CA2331934A1 - Protein and its encoding nucleotide sequences for diagnosis, prevention and treatment of lung injuries and disorders - Google Patents

Abstract

The present invention describes the identification of a novel protein P-SCGB
belonging to the family of uteroglobins/secretoglobins, and nucleic acid sequences encoding said protein. Said protein and nucleic add sequences are useful in the diagnosis and/or treatment of lung injuries and diseases, and of inflammation or immunomodulation-related disorders. More particularly said protein and nucleic add sequences are useful in the diagnosis and/or treatment of asthma. The new protein is specific for the lung and has a small size facilitating its transepithelial leakage from the lung into serum. It therefore has all the features to serve as a peripheral lung marker.
Furthermore, its homology with members of the uteroglobin/secretoglobin family make it a potentially important protein that may be used as a diagnostic or therapeutic tool for many other pathologies.

Description

Protein and its encoding nucleotide sequences for diagnosis, prevention and treatment of lung injuries and disorders FIELD OF THE INVENTION
The present invention relates to a novel protein, named pneumo-secretoglobin (P-SCGB), nucleic acid sequences encoding said protein and portions thereof.
The invention further relates to the use of said nucleic acids or protein sequences as a marker of respiratory health status and its use in the prognosis, diagnosis, prevention and/or treatment of lung ~njuries and diseases, and of inflammation or immunomodulation-related disorders. More particularly, the present invention relates to the use of said nucleic acids or proteins for diagnosis or treatment of asthma. The present invention also relates to the use of the spec'rfic marker in the assessment of a compound's impact on trachea and lung integrity.
BACKGROUND OF THE INVENTON
It is known that the lung interfaces with the environment access a continuous and heterogeneous epithelium. The proximal conducting airways are lined with a pseudostratified epithelium that is progressively replaced by a simple cuboidal cell layer in the more distal airways and by a very thin epithelial lining coating more than 95% cf the lung's surface area in the alveoli. Numerous cell types, irxluding ciliated, basal, goblet and Clara cells, are present along the airways (Breeze R. G. et al., Am. Rev.
Respir. Dis. 116:705, 1977).
The primary function of the lung and of the alveolar epithelium in particular, is to provide an extensive and thin surface for gas exchange. The pulmonary epithelium serves a number of additional functions that basically act to preserve the capacity for such gas exchange. It provides a barrier that protects the host from the outside environment by segregating inhaled foreign agents, and it controls the movement of solutes and water, contributing 'o the maintenance of lung fluid balance. The lung epithelium also plays an active role in the metabolism of endogenous mediators and xenobiotic agents, and is capable of regeneration, allowing normal cell turnover and restoration of airway and alveolar functions after lung injury. Beyond this, the lung epithelium produces complex secretions, among which is the mucus blanket, a surface-active agent (surfactant), as well as several proteins important for host defense (Shale S.
et al., In R.G. Crystal, J.B. West, E.R. Weibel and P.J. flames, ed. The Lung:Sdentific Foundations. Lippincotl-Raven, Philadelphia, 479, 1996).
Sampling of the epithelial lining fluid (ELF} by bronchoalveolar lavage (BAL) is the common means of studying the proteins secreted by the lung epithelium and investigating their changes in lung diseases (Reynolds H. Y. and Newball H.H., J Lab.
Clin. Med. 84:559, 1974}. Recent studies have shown that some proteins secreted by airvvay or alveolar epithelial cells are present not only at the surface of the respiratory tract, but also and normally in small amounts in the bloodstream. Among these proteins to are the 16-kD Clara cell secretory protein (CC16, CC10), three surfactant-associated proteins (surfactant protein [SP]-A, SP-8, and SP-D) and mucin-associated antigens, as recognized by monoclonal antibodies (KL-6, 17-81, 17-D2). Because these proteins are mainly, if not exclusively, secreted within the respiratory tract, their occurrence in the vascular compartment is explained by their leakage from the lung into the bloodstream.
The leakage of proteins from the lung into the circulation appears to be governed by a number of factors, among which the permeabiiityof the tight bronchoalveolar epithelium, and the molecular features of the proteins themselves (e.g., size and charge) and their exchangeable pool, appear to be critical (Hermans C. et al., Am. J. Respir.
Crit. Care Med. 159(2):646, 1999). The significance of the levels of these proteins in serum 2o depends markedly on whether the air-blood barrier is intact or damaged.
Moreover, these proteins show variations in the serum of patients with d'rfferent lung diseases and subjects exposed to lung toxicants (Lindaht M. et al., Thorax 51:1028, 1996;
Lenz A. G.
et al., Electrophoresis 11:540, 1990; Lenz A.G. et aL, Electrophoresis 14:242, 199.
Patent application W099/09054 discloses the discovery of a new bronchoalveolar protein, belonging to the family of peroxiredoxins. The inventors identified proteins from bronchoalveolar lavage fluids (BALF's) which were obtained by washing the epithelial lining fluid of the lungs with saline. A pool of BALFs from various patients was analyzed by 2D electrophoresis and was fcund to contain a total of 211 silver-stained spots in the range of pl 3.5-10 and molecular mass 5-100 kDa. Next, 182 spots were identrfied by microsequence analysis and by matching with human blood plasma and the Macrophage Like Cell line reference 2-DE maps available from the SWISS-2DPAGE
database. The human bronchoalveolar lavage fluid was finally found to contain different proteins or isoforms thereof. Most of the proteins identified in the lavage were known and either were produced locally or originate from plasma. However, one particular peptide sequence did not correspond to a previously know protein and thus was subjected to further analysis to unravel the potentially new protein associated. The DNA coding sequence was cloned and sequenced and the protein was found to be a new peroxiredoxin (Knoops B. ef al., J. Biol. Chem. 274(43):3045 t, 1999).
As mentioned above, the concentration of lung specrfic proteins in serum can be used in the evaluation of lung disorders with a similar utility as proteinuria in kidney diseases involving the glomeruli. Human and experimental data indicate that comparable structural and functional features govern the passage of proteins across the lung epitheliumJblood barrier and the glomerular filter. ft is known that the concentrations in 1o serum of some lung-specific secretory proteins, such as the bronchiolar Clara cell 16 kD
protein (CC16) and alveolar surfactant-associated proteins A and 8 (SP-A and SP-B), can be used to assess the integrity of the respiratory tract epithelium.
Therefore, newly identified lung-specific proteins could be used as diagnostic tools for lung disorders in the same way as human Clara cell protein (CC16, another member of the uteroglobin/secretoglobin family). In addition, such newly identified molecules may help to develop and/or to improve several therapeutic applications of various lung injuries and disorders, and of inflammation- or immunomodulation-related disorders. Any protein spec'rfic for the lung and secreted in respiratory airways is likely to be used as peripheral lung marker especially if it is a small size protein (molecular 2o weight less than 20 kD) readily diffusible from the lung into serum.
Therefore, it is a major goal of the present invention to provide nucleic acid as well as amino acid sequences encoding novel lung-specific proteins.
It is another major goal of the present invention to provide methods and kits for the diagnosis and detection of pulmonary injuries and disorders, as well as inflammation or immunomodulation-related disorders. Human health is determined by the complex interplay between genetic susceptibility and environmental exposure. It is a further goal of the invention to demonstrate the linkage of the new protein and sequences to lung diseases and disorders, as well as inflammation- or immunomodulation-related disorders, and to provide tools for the analysis of such disorders. In particular the 3o determination of an individual's allelic pattern for a gene related to particular diseases provides greater insight into the identification of individuals at risk for such diseases, early prevention and therapy, and understanding of the pathology. It is a further goal of the invention to provide methods for this application in the field of lung diseases and disorders, as well as inflammation- or immunomodulation-related disorders.

It is a further goal of the present invention to provide compos~fions for the prevention andlor treatment of said lung injuries and the related disarde.~s as metrsoned above.
Another goal of the present invention is to provide methods a>tvw to screen ccrnpounds used for the prevention or treatment of lung injuries and reed deso~s.
All the goals of the present invention have been met by the emooc5rnertts at set a.t below in the description of the invention.
SJMMARY OF THE INVENTION
to The essence of this invention is based on the identif'~ ~ a never low molecular weight human bronchoalveolar protein and its encodes reudeic acid sequence. Said protein was found by studying 2D electrophoresis ~oten reaps of h;~man bronchoalveolar lavage fluids (BALF's) from individual patients w~ wNl-defined t5 interstitial lung diseases (sarcoidosis, idiopathic pulmonary fps (1PF] and hypersensitivity pneumonitis (HP)). The protein spots were furtt~ analysEd by r~icrosequence analysis and by matching with human blood ~asr-~a and the t~!$crophage Like Cell Line reference 2-DE maps available from tte S~NtSS-2D
PAGE
database. This microsequencing gave the sequence of a short pence from which 20 o~gonucleotides were designed to deduce the sequence of the compi~ txxr~an cDNA
~quence (SEQ ID N02) of a new bronchoalveolar protein (SEQ i0 ~t0 t), named pneumo-secretoglobin (P-SCGB). In addition, the human genomic DNA seq;~ence (SEo ID NO 3), and the homologous mouse cDNA (SEA 10 NO 4) and its encoring amino acid sequence (SEO ID NO 5) have been revealed. It was furthem~ap demonstrated 25 tt'~at the expression of said protein is very specific to the respiratory tract, esa. l4ng and t~achea.
DETAILED DESCRIPTION OF THE INVENTION
3o According to a first embodiment, the present invention relates to an ~aated ~~ucleic acid encoding a new lung marker protein or an immunologocaA~f acwe andlor a '~nctional fragment of said protein selected from the group consistasg of ;a) a nucleic acid comprising or consisting of at least a functional ,:art of the DNA
sequence as given in SEQ ID NO 2, 3 or 4, or the complement thefxt.

(b) a nucleic acid which selectively hybridizes with any of the sequences as given in SEO 10 NO 2, 3, 4, 7, 8, 9 or 10, (c) a nucleic acid comprising at least a functional part of a sequence encoding a protein with an amino acid sequence which is at least 60 ~ homologous to the amino acid 5 sequence as given in SEA ID NO 1 or 5, (d) a nucleic acid encoding a protein comprising or consisting of the amino acid sequence as given in SECT ID NO 1 or 5, (e) a nucleic acid sequence which is degenerated as a result of the genetic code to a nucleotide sequence encoding a protein as given in SEO NO 1 or 5 or to a nucleic 1o acid as defined in (a) to (d), and, (t) a nucleic acid sequence encoding a protein as defined in SEQ NO 1 or S or as defined in any one of (a) to (e) interrupted by intervening DNA sequences.
As illustrated in Example t , the present inventors were able to clone and sequence a new gene encoding a new human bronchoalveolar protein, named pneumo secretoglobin (P-SCGB). This new protein was found by sequencing proteins from human bronchoalveolar fluids (BALF's) which were separated on a 2D-electrophoresis get. The present inventors surprisingly found that this until now unknown protein was differentially expressed in some lung pathologies, as it clearly appears in bronchoalveolar lavage fluids (BALF) from patients suffering from idiopathic pulmonary fibrosis (1PF} and is almost undetectable in BALF from controls.
In Figure 1 the cDNA sequence given by SEA ID NO 2 and the corresponding amino acid sequence given by SEA ID NO 1 is represented. Sequence similarity searches on both nucleic acid (SEO ID NO 2) as protein level (SEQ ID NO 1 ) with sequences available in public databases revealed that the cDNA sequence including its encoded protein sequence do not share significant homology with previously known proteins or cDNA's of human origin, and therefore can be defined as new.
According to the invention, the protein P-SCGB given by SEG1 ID NO 1 presents a pl of 7.66 and a molecular weight of 10161 D. Other structural properties of P-SCGB
are further defined in example 2. The sequence of this new protein given by SEO ID NO
1 shows sign'rficant similarity with the consensus sequence of the uteroglobins/secretoglobins, as illustrated in example 3. This finding together with additional functional properties of the protein as described in example 3 indicate that the new protein P-SCGB can be classified within the "uteroglobin/secretoglobin"
family. The inventors were also able to clone the homologous gene from mouse (example 5).
In Figure 5 the mouse cDNA sequence given by SEO ID NO 4 and its corresponding amino acid sequence given by SEQ ID NO 5 are represented. Finally, the inventors have also identified the genomic sequence of P-SCGB (example 6). This genomic sequence is given by SEA ID NO 3 and illustrated in Fgure 7.
The term "nucleic acid" as used above refers to DNA or RNA, or ampl'rfied versions thereof, or the complement thereof. DNA may refer to genomic, synthetic DNA
or cONA.
The term "isolated" distinguishes the protein or nucleic aad according to the invention from the naturally occuring versions thereof. A nucleic acid accorcf~ng to the to invention may also comprise any modified nucleotide known in the art.
The term "complement' refers to a nucleotide sequence which is complementary to an indicated sequence and which is able to hybridize to the indicated sequence.
The term "functional fragment" when referred to nucleic acids, refers to a part of the nucleic acid encoding substantially at least one biological function of the protein which said nucleic acid is encoding.
In the context of the present invention, and since P-SCGB is specifically expressed in the airway tract, it is believed that this protein is very important in the field of lung and/or inflammation related disorders. A list of lung disorders which may be related to the expression of P-SCGB is given ~n the description of this invention further 2o below.
Furthermore, the inventors could assign the gene coding for the protein P-SCGB
to chromosome 5 locus Sq31-33 (example 6). Therefore P-SCGB is likely to play a role in diseases linked to this chromosomal locus. Evidence for linkage of this locus to the immune response was provided by studies showing that it is a site of regulation of peripheral blood eosinophilia (Martinez F.D. ef al., Am. J. Respir. Crit. Care Med.
158(6):1739, 1998) and the immunoglobulin E concentration in serum. These parameters correlate with clinical expression of allergy, atopy and asthma.
More precisely, this chromosomal locus has been associated with disorders of the bronchial responsiveness. Indeed, chromosome 5 is particularly studied in the field of asthma research. Asthma is known as a complex heritable disease. This means that there are a number of genes that contribute towards a person's susceptibility to that disease, and in the case of asthma, chromosomes 5, fi, 11, 14 and 12 have all been implicated.
The relative roles of these genes in asthma predisposition are not clear, but one of the most promising sites for investigation is on chromosome ~. Although a gene for asthma from this site has not yet been specifically identified, it is known that this region is rich in genes coding for key molecules in the inflammatory response seen in asthma.
This is consistent with the suspected anti-inflammatory properties of P-SCGB.
Loci associated with asthma have been more precisely located to 5q31-33 and 11 q13 (Palmer L.J. et al., Am. J. Respir. Crit. Care Med. 158(6):1825, 1998).
It is known that 11q13 is the locus of CC16 (Zhang Z. et al., DNA Cell Biol. Jan 16(1):73.
199, a protein definitely associated with asthma (Lensmar C. et al., Ce!! Mol. Life Sci.
57(6):976, 2000), and it is known that a mutation in the CC16 gene is associated with asthma (Laing LA. et al., J. Med. Genet. 35(6):463, 1998).
to The fact that the chromosomal locus for P-SCGB is linked to asthma, like the one for CC16, the fact that P-SCGB is an uteroglobin-like, small MW, secreted and lung-specific protein like CC16, lead the inventors to suggest that P-SCGB is associated with asthma. Further evidence for linkage of the chromosomal linkage of the chromosomal locus of P-SCGB to asthma and disorders of responsiveness is provided by numerous studies (Los H. et al., Eur. E~espir. J. 14(5):1210, 1999; Mansur A.H. et aL, Clin. Exp.
Allergy 28(2):141, 1998; Holgate S. T., J. Allergy Clin. ImmunoL 104(6):1139, 1999; Km H. S. et al., Cun-. Opin. Pulm. Med. 4(1)'46, 1998; Mansur A.H. et al., CGn.
Exp. Allergy 28(2):141, 1998; Kawakami Y. et aL, Respirology 2(1):7, 1997).
It has also been shown that chromosome 5 aberrations are associated with an Zo increased risk for lung cancer (Wu X. et al., Int. J. Cancer 79(5):490, 1998), or an increase in prostate cancer aggressiveness (Sark T et al., Int. J. Cancer 81(2):219, 1999) the locus being once again 5q. This is consistent with some properties already demonstrated for CC16, i.e. that its overexpression reduces carcinogenesis and has tumor suppressor-like effects (Zhang Z. et al., Pros. Natl. Acad. Sci. USA
96(7):3963, 1999). Thus information provided in the present invention are useful in the prognosis, diagnosis and/or treatment of some forms of cancer, more particularly prostate and lung cancer.
The term "functional fragment" when referred to peptides or proteins, refers to a fragment having substantially at least one of the biological activities of the polypeptide from which it is derived. The term "functional fragment" of a protein relates to a truncated version of the original protein or polypeptide referred to. The truncated protein sequence can vary widely in length; the minimum size being a sequence of sufficient size to provide a sequence with at least a comparable function and/or activity of the original sequence referred to, while the maximum size is not critical. In some applications, the maximum size usually is not substantially greater than that required to provide the desired activity and/or functions) of the original sequence. A functional fragment can also relate to a subunit with similar function as said protein. Typically, the truncated amino acid sequence will range from about 5 to about 60 amino acids in length.
More typically, however, the sequence will be a maximum of about 50 amino acids in length, preferably a maximum of about 60 amino acids. It is usually desirable to select sequences of at least about 10, 12 or 15 amino acids.
Functional fragments include those comprising an epitope or an amino acid stretch which is specific or unique for the proteins according to the invention. Epitopes may be determined using, for example, peptide scanning techniques as described in Geysen et al. (1996). Preferred functional fragments have a length of at least, for example, 5, 10, 25 or 50 amino acids.
With "immunologically active' is meant that a molecule or specific fragments thereof such as epitopes or haptens are recognized by, i.e. bind to antibodies. This i5 recognition is preferably a specific immunological recognition.
The term 'spec'rfically hybridizing" means hybridizing under conditions wherein sequences can be detected which are homologues of the sequences of the invention, but which are for instance derived from heterologo~s cells or organisms, and wherein said sequences do not hybridize with known sequences. It is well known to the person skilled in the art which methods for hybridization can be used and which conditions are necessary for selectively or specifically hybridizing. Hybridization conditions are essentially described in Sambrook (SambrookJ., Fritsch E.F. and Maniatis T.
Molecular cloning: a laboratory manual. Cold Spring Harfwr Laboratory Press, blew York;
198, and preferably specific or high stringently hybridization conditions are aimed at. Stringent conditions are sequence dependent and will be ditierent under different circumstances.
Generally, stringent conditions are selected to be about 5°C lower than the thermal melting point Tm for the specific sequence at a defined ionic strength and pH.
The Tm is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. Typically, stringent conditions will be 3o those in which the salt concentration is about 0.02 molar at pH 7 and the temperature is at least about 60'C.
According to a more specific embodiment, the present invention relates to a nucleic acid sequence which is at least 35 % identical to the nucleic acid sequence as given in SEC7 ID NO 2. Preferably said nucleic acid is 40 %, 45 % or 50 %
identical, more preferably 55 °~, 60 %, 65 %, 70%, 75°~6, 80%, 85%, 90%, most preferable 95 identical to the nucleic acid sequence as given in SEO ID NO 2.
According to another specific embodiment, the present invention relates to a nucleic acid sequence which is at least 35 % identical to the nucleic acid sequence as given in SE(7 ID NO 3. Preferably said nucleic acid is 40 %, 45 % a 50 %
identical, more preferably 55 %, 60 %, 65 %, 70%, 75%, 80%, 85%, 90°~6, most preferable 95 identical to the nucleic acid sequence as given in SEO ID NO 3.
According to another specific embodiment, the present invention relates to a nucleic acid sequence which is at least 35 % identical to the nucleic acid sequence as io given in SEA ID NO 4. Preferably said nucleic acid is 40 °~, 45 % of 50 % identical, more preferably 55 %, 60 %, 65 %, 70%, 75%, 80%, 85%, 90%, most preferable 95 identical to the nucleic acid sequence as given in SEQ ID NO 4.
According to yet another embodiment the present invention relates to a nucleic acid sequence comprising a sequence encoding a protein which is at least 60 i5 homologous, preferably 65%, 70 % or 75 % homologous, more preferable 80 %, 85 % or 90 % identical, most preferable 95 % homologous to the amino acid sequences as given in SEO tD NO 1.
According to yet another embodiment the present invention relates to a nucleic acid sequence comprising a sequence encoding a protein which is at least 60 2o homologous, preferably 65%, 70 % or 75 % homologous, more preferable 80 %, 85 % or 90 % identical, most preferable 95 % homologous to the amino acid sequences as given in SEA ID NO 5.
The term "identityl' as used above means the degree of sequence relatedness between two polypeptide or nucleic acid sequences as determined by the identity of the 25 match between two strings of such sequence. The term "homology' between two polypeptides is determined by comparing the amino acid sequence of one polypeptide to the sequence of a second polypeptide. Amino acids are considered homologous when they have the same basic properties in view of their hydrophobicity, hydrophilicity, hydrophobic moment, etc. (Vogr G. ef ai., J. Mol. Biol. 249(4):816, 1995).
30 As a practical matter, whether any particular amino acid or nucleic acid sequence is identical or homologous to, for instance, the amino acid or nucleic acid sequence of this invention, can be determined conventionally using known computer program methods. Such computer program methods include, but are not limited to, GCG

program package (Oevereux J., Nucleic Acids Research 12, 387; 1984), BLASTP, BLASTN and FASTA (Alfschul S.F., et al., J. Mol. Biol. 215, 403; 19907.
The present invention is also 'related to an antisense molecule comprising a nucleic acid sequence capable of specifically hybridizing to a nucleic acid of the present invention as defined above. Such antisense molecule allows to block a nucleic acid sequence of the invention. Nucleic acids of the invention may be inserted into vectors in an antisense orientation to provide far the production of antisense RNA. In addition, antisense RNA or other antisense nucleic acids may also be produced by synthetic means.
to Said antisense molecules may be particularly advantageous for development of therapeutics for lung or related disorders. For example, an antisense nucleic acid capable of binding to the nucleic acid sequences of the invention may be used to selectively inhibit expression of the corresponding polypeptides, leading to reduction of associated lung disorders or related disorders.
IS According to further embodiments the present invention also relates to probes or primers containing a sequence of at least 15 contiguous nucleotides of a nucleic acid as defined above.
The probes will hybridize specifically with any of the ~ nucleic acids of the invention.
zo The term "probe" according to the present invention refers to a single-stranded oligonucleotide sequence which is designed to seleCively hybridize to any of the nucleic acids of the invention. The probes used in the process of the invention can be produced by any method known in the art, such as cloning of recombinant plasmids containing inserts including the corresponding nucleotide sequences, rf need be, by cleaving the z5 latter out from the Boned plasmids upon using the appropriate nucleases and recovering them (e.g. by fractionation according to molecular weight). Preferably the probe is about 5-50 nucleotides, more preferably 20, 25, 30, 35, 40 or 45 nucleotides.
The primers will specifically amplify any of the nucleic acids of the invention.
The term "primer" refers to a single stranded nucleotide sequence capable of 3o acting as a point of initiation for synthesis of a primer extension product which is complementary to the nucleic acid strand to be copied. The length and the sequence of the primer must be such that they allow to prime the synthesis of the extension products.
Preferably the primer is about 5-50 nucleotides, more preferably 20, 25, 30, 35, 40 or 45 nucleotides. Specific length and sequence wilt depend on the complexity or the required it DNA or RNA targets, as well as on the conditions of primer use such as temperature and ionic strength. The fact that amplification primers do not have to match exactly with corresponding template sequence to warrant proper amplification is amply documented in the literature (Kwok et al., Nucl. Acids Res. 18:99, 1990).
The primers according to the invention may be used in polymerise chain reaction (PCR) cloning mechanisms which generally involve making a pair of primers, which may be from approximately 10 to 50 nuGeotides to a region of the gene which is desired to be cloned, bringing the primers into contact with mRNA, cDNA or genomic DNA to be amplified, performing a polymerise chain reaction under conditions which 1o bring about ampf'rfication of the desired region, isolating the amplified region or fragment and recovering the amplified DNA. Generally, such techniques as defined herein are well known in the art, such as described in Sambrook (Sambrook J., Frttsch E.F. and Maniatis T. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York; 1989). These primers can also be used to clone homologues of the nucleic acid sequences of the invention in other organisms.
The probes and primers as defined above can also be synthesized chemically, for instance, by the conventional phopho-triester method.
The probes and primers of the invention can optionally be labeler using any canventional label. ~ his may include the use of labelled nucleotides incorporated during the polymerise step of the amplification or by any other method known to the person skilled in the art. Suitable labels include radioisotopes such as ~P, ~'P
or'~S, enryme labels or other protein labels such as biotin or fluorescent markers. Such labels may be added to any nucleic acid or oligonucleotide of the invention and may be detected using techniques known in the art.
The present irivention is also related to a vector comprising a nucleic acid sequence as defined above.
Additionally, this vector may be transformed, transfected or infected into a host cell.
According to a more preferred embodiment, said vector is an expression vector 3o comprising one or more adjacent regulatory or control sequences (such as promoter(s), secretion and termination signal sequence(s)) advantageously operably Inked to the nucleic acid sequence of the invention.
The vector according to the invention may advantageously be a plasmid, cosmid, virus or other suitable vector which is known to those skilled in the art.
E~ression can be done in both prokaryotic as eukaryotic host cells such as bacteria, yeast, fungi, insect or mammalian cells, which have been previously transformed or transfected by a vector coding for the polypeptides of the invention. It is expected that those of skilled in the art are knowledgeable in the numerous expression systems available for expression in these systems.
According to another embodiment, the present invention relates to an isolated new lung marker protein comprising one of the polypeptides selected from the group consisting of (a) a polypeptide as given in SEO ID NO 1 or 5, (b) a polypeptide with an amino acid sequence which is at least 60 %
homologous to the amino acid sequence as given in SEO ID NO 1 or 5, (c) a polypeptide encoded by a nucleic acid as given in any of SEO ID NO 2,3 or 4, (d) a polypeptide encoded by a nucleic acid as given in any of claims 1 or 2, or a homologue or a derivative of said protein, or an immunologically active andlor functional fragment thereof.
The term "protein" in the context of the present invention is used interohangeabty with the terms 'polypeptide" and "peptide".
The term "hcmologue" of a protein of the invention are those peptides, oligopeptides, polypeptides, proteins and enzymes which contain amino acid substitutions, deletions and/or additions relative tv the said protein with respect to which they are a homolog, without altering one or more of its funcfional properties, in particular without reducing the activity of the resulting. For example, a homolog of said protein will consist of a bioactive amino acid sequence variant of said protein. To produce such homologs, amino acids present in the said protein can be replaced by other amino acids z5 having similar properties, for example hydrophobicity, hydrophilicity, hydrophobic moment, antigenicity, propensity to form or break a-helical structures or ~-sheet structures, etc.
The term "derivative" of a protein of the invention are those peptides, oligopeptides, polypeptides, proteins and enzymes which comprise at least about five or ten contiguous amino acid residues of said polypeptide but which retain the biological activity of said protein. A "derivative" may further comprise additional naturally-occurring, altered glycosylated, acylated or non-naturally occurring amino acid residues compared to the amino acid sequence of a naturally-occurring form of said polypeptide.
Alternatively or in addition, a derivative may comprise one or more non-amino acid t3 substituents compared to the amino acid sequence of a naturally-occurring form of said polypeptide, for example a reporter molecule or other ligand, covalently or non-covalently bound to the amino acid sequence such as, for example, a reporter molecule which is bound thereto to facilitate its detection.
Said polypeptide of the invention may be produced in a recombinant manner The present invention therefore relates to a method for producing a polypeptide as defined above comprising culturing a host cell under conditions allowing the expression of said polypeptide and recovering said produced polypeptide from the culture.
Recombinant host cells according to the present invention as described below may be used for the 1o recombinant expression of said polypeptide.
The polypeptide may be expressed in a modified form, such as a fusion protein, and may include not only secretion signals, but also additional heteroiogous functional regions. For instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistance in the host cell, during purification, or during subsequent handling and storage. Also, peptide moieties may be added to the polypeptide to facilitate purification.
Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to polypeptides to engender secretion or excretion, to improve stability and facilitate purification, among others, are familiar and routine techniques in 2o the art.
Said recombinant polypeptides may advantageously be used in diagnostic methods or kits or the like.
According to yet another embodiment the present invention is related to an antibody specifically recognizing a protein as defined above, or recognizing 's immunologically active parts or specific epitopes thereof.
The term "antibody" refers to both monoclonal or polycfonal antibodies, capable of specrfically binding to one or more epitopes of the proteins of the invention.
The term "specifically recognizing' implies that there is substantially no cross reaction of the antibody with other proteins. The antibodies according to the invention 3o may be produced according to techniques which are known to those skilled in the art.
Monoclonal antibodies may be prepared using conventional hybridoma technology as described by Kohler and Milstein (Kohler F. and Milstein C. Nature 256, 495;
1975). This classical method comprises producing any hybridoma formed by, on the one hand, isolating splenic lymphocytes of an animal, particularly a mouse or a rat immunized against a polypeptide of the present invention or a fragment as defined above, and cells of a myeloma cell line on the other hand, and selecting said hybridoma for the ability to produce the monbclonal antibodies recognizing the polypeptide which has been initially used for the immunization of the animals.
The antibodies involved in the invention can be labelled by an appropriate label of the enzymatic, fluorescent or radioactive type or any label known in the art.
Alternative to preparing monoclonal antibody-secreting hybridomas, a monoclonal antibody directed against a polypeptide of the invention can be identified and isolated by screening a recombinant combinatorial immunoglobuiin library (e. g., an to antibody phage display library) with the polypeptide of interest. flits for generating and screening phage display libraries are commercially available (e. g., the Pharmacia Recombinant Phage Antibody S~sie~rn, Catalog No. 27-9400-01; and the Stratagene SurJZAPPhage Display Krt, Catalog No. 240612). Additionally, examples of methods and reagents particularly amenable for use in generating and screening antibody display IS library can be found in, for example, U. S. Patent No. 5,223,409; PCT
Publication No.
WO 92/18619; PCT Publication No. WO 91/17271; PCT Publication No. WO 92/20791;
PCT Publication No. NlO 92/15679; PCT Publication No. WO 93/01288; PCT
Publication No. WO 92/01047; PCT Publication No. WO 9?J09690; PCT Publication No. WO 90/02809; Fucks et al. BioITechnology 9: 1370, 1991; Hay et at.
20 Hum. Anfibod. Hybridomas 3: 81, 1992; Huse et al. Saence 246: 1275, 1989;
Griffiths et al. EMBO J. 12: 725, 1993).
Additionally recombinant antibodies, such as chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, which can be made using standard recombinant DNA techniques, are within the scope of the z5 invention.
A chimeric antibody is a molecule in which different portions are derived from different animal species, such as those having a variable region derived from a murine mAb and a human immunoglobulin constant region. (see, e. g., Cabilly et al., U. S.
Patent No. 4,816,567; and Boss ef al., U. S. Pafenf No. 4,816397, which are 3o incorporated herein by reference in their entirety.) Humanized antibodies are antibody molecules from non-human species having one or more complementarily determining regions (CDRs) from the non-human species and a framework region from a human immunoglobulin molecule (see, e. g., queen, U.
S. Patent No. 5,585,089, which is incorporated herein by reference in its entirety.) Such IS
chimeric and humanized monoclonal antibodies can be produced by recombinant DNA
techniques known in the art, for example, but not limited to, using methods described in PCT Publication No. WO 87/02671; European Patent Application 184,187; European Patent Application 171,496; European Patent Application 173,494; PCT
Publication No.
WO 86/01533; U. S. Patent No. 4,816,567; European Patent Application 125,023;
Better et al. Science 240: 1041, 1988; Liu et aL Proc. Natl. Acad. Sci. USA 84: 3439, 1987; Liu et al. J Immunol. 139: 3521, 1987; Sun et al. Proc. NatL
Acad. Sci. USA 84: 214, 1987; Nishimura et al. Canc. Res. 4T 999, 1987; Wood et aL
Nature 314: 446, 1985; Shaw et al. J. Natl. Cancer Inst.
80: 1553, 1988; Marrison Science 229: 1202, 1985; Oi et al. SiolTechniques 4:
214, 1986; U. S. Patent 5,225,539; Jones et al. Nature 32i: 552, 1986; Verhoeyan et al.
Science 239: 1534, 1988; Beidler et al. J. lmmunol. 141: 4053, 1988; Holliger et al.
Cancer Metastasis Rev. 18(4):411, f 999; and Kipriyanov et al., 12(2):173, 1999).
Completely human antibodies are particularly desirable for therapeutic treatment t5 of human patients. Such antibodies can be produced, for example, using transgenic mice which are incapable of expressing endogenous immunoglobulin heavy and light chains genes, but which can express human heavy and light chain genes. The transgenic mice are immunized in the normal fashion with a selected antigen, e. g., all or a portion of a polypeptide of the invention. Monoclonal antibodies directed against the antigen can be obtained using conventional hybridoma technology. The human immunoglobulin transgenes harbored by the transgenic mice rearrange during B
cell differentiation, and subsequently undergo class switching and somatic mutation. Thus, using such a technique, it is possible to produce therapeutically useful IgG, IgA and IgE
antibodies.
For an overview of this technology for producing human antibodies, see Lonberg and Huszar (Int. Rev. Immunol. 13: 65-93, 1995). For a detailed discussion of this technology for producing human antibodies and protocols for producing such antibodies, see, e. g., U. S. Patent 5,625,126; U. S. Patent 5,633,425; U. S. Patent 5,569,825; U. S.
Patent 5,661,016; and U. S. Patent 5,545,806. In addition, companies such as Abgenix, Inc. (Fremont, CA), can be engaged to provide human antibodies directed against a selected antigen using technology similar to that described above. Completely human antibodies which recognize a selected epitope can be generated using a technique referred to as 'guided selection." In this approach a selected non-human monoclonal antibody, e. g., a mouse antibody, is used to guide the selection of a completely human l6 antibody recognizing the same epitope. (Jaspers ef al.
&otechnology 12: 899, 199.
Polyclonal antibodies may also be prepared using conventional technology well known to those skilled in the art, and which comprises inoculating a host animal, such as a mouse, with a protein or epitope according to the invention and recovering the immune serum.
The present invention also includes fragmertis of whole antibodies which maintain their binding activity, such as for example, Fv, F(ab') and F(ab')2 fragments as well as single chain antibodies, and which are prepared according to methods weU
l0 known in the art.
Antibodies according to the invention may be used in a method of detecting the presence of a polypeptide according to the invention, which method comprises reacting the antibody with a sample from a body fluid (of a patient) and identifying any protein bound to said antibody. A diagnostic kit may also be provided for performing said method which comprises an antibody according to the invention and means for reacting the antibody with said sample.
The antibodies according ;o the invention may also be used as a medicament for treating lung disorders, or may be ~mprised in a pharmaceutical composition.
According to a further embodiment, the present invention also relates to an arrfi-idiotype antibody raised against the antibody as defined above.
In particular, an anti-idiotype antibody raised against an antibody as defned above, refers to a monoclonal antibodies raised against the antigenic determinants of the variable region of the antibody raised against the polypeptides of the present inventron or against fragments thereof as defined above. These antigenic determinants of immunoglobins are known as idiotypes (set of idiotypes) and can therefore be considered to be the "fingerprint of an antibody. Monoclonal anti-idiotypic antibodies have the property of forming an immunological complex with the idiotype of the monoclonal antibody against which they were raised.
In this respect the monoclonal antibody is referred to as Abl, and the anti idiotypic antibody is referred to as Ab2. Ab2 or fragments thereof can thus recognise the interaction site or receptor of ;he polypeptide of the present invention, and can consequently block this interaction site. As a result Ab2 may prevent any lung disorder associated with an interaction between the polypeptide and its receptor. These anti idiotype antibodies can be prepared by methods well known in the art.

t~
Anti-idiotypic antibodies may also be used in c ats ,.sed for ~e detection of lung and related disorders.
According to a next embodiment the present in~~: is .~eiat$d ~ a nor~-~s-man transgenic animal transformed by a nucleic acid as defines above, or ~r a ~ ~
~e present invention as defined above.
In a more preferred embodiment, the present investor rela;.es oc a me>~soa for the production of a genetically modified non-human anrrat in wryd's ~ s modficaZzon results in overexpression, underexpression or no expresson of Lne nrdac aac~
or the polypeptides of the invention as defined above.
1o Said anima( is preferably a mammal such as a ma.se or a ra: trar'sfom'ed by a vector according to the invention and overexpressing a protein ac~rding >3o the invention, or genetically modified by a partial or total ~ ~~ ~~~e encoding the protein according to the invention (a knodc~u; non-hurrsar:
~~ar'~ct~1 and obtained by methods well known by the person skilled in the ar. sr~ as ='ze one t5 described by Kahn et al. (Cell 92: 593596; 1998.
In particular, the present inventors have ider~e~d ~e h~-~obc;:.us «cuse sequence of P-SCGB, as further illustrated in example i Ktcvwecge ~ d'ns '''~cuse sequence is the first and necessary step in the generation ~ ~ransges-~c a~
~'aa.
t7ther examples of genetically modified non-hcrar ar,mais :.r'~~d~ ~ fi''e 2o invention are for instance transgenic non-human arur~s comprssr~ ar a~sense sequence as defined above and complementary to rie ~e~c a-=~ ~en~ces according to the invention, and placed in such a way tha_ ~ is ~ns~'~ ~'I"~
3_~s~ense mRNA which is complementary to the nucleic acid sequ~es assorting b ~e in'~~~ti°n and which hybridises to said nucleic acid sequences, the~Y ~~r~9 ~
b~'"°r'9 ~e'r 25 translation.
A further aspect of the present invention is relates ~n a com~o~r' 's~9 at least one nucleic acid sequence, an antisense molecule, 3 pot~peptfce- ar, artt~c~r, an anti-idiotypic antibody as defined above, optionally in acr,-~x'u'e Win" a ;~arma~"~t tally acceptable carrier.
3o A pharmaceutically acceptable carrier includes a°~ ca er ti~i yes r:~t itself induce the production of antibodies harmful to the indi~r~a.:a~ 'ece~Y~g ='~e ~r"ion.
Suitable carriers are typically large, slowly metabol~z:~g rr'aa"..r'ra°~..les sLCi as proteins, polysaccharides, polylactic acids, polyglycouc ands. ~YT"'~'~ ~'~
aids, amino acid copolymers; and inactive virus particles. S..aw~~ers a,- weif to I$
those of ordinary skilled in the art. Pharmaceutically acceptable carriers vary accorang to the mode of administration (intravenous, intramuscular, subcutaneous, parental, etc.), and may comprise also adjuvants well known by the person skilled in the art to increase, reduce andlor regulate humoral, local andlor cellular response of the immune system.
According to a further embodiment said composition may be used as a medicament.
In a more preferred embodiment, said medicament is used for the diagnosis, prevention andlor treatment of a patient, especially a human patient, preferably affected by lung disorders or any related disorder as defined hereunder. A sufficient amount of the pharmaceutical composition is administered to said patient in order to detect, trzat, avoid and/or reduce the symptoms of said injuries and/or diseases.
The various types of lung disorders which may be treated according to the present invention can subdivided within 3 groups: (1 ) lung disorders caused by altered t5 air/blood (or alveolocapillary) permeability e.g. sarcoidosis, fibrosis, acute respira;ory distress syndrome CARDS), oedema, lung injury caused by toxic chemicals (omne, particles, chlorine) or particles (crystalline silica, dust) and lung inflammation (bronc~rtis, pneumonia, LPS (lipopolysaccharides); (2) lung disorders caused by acute or chronic epithelium damage e.g. BPCO (bronchopulmonary chronic obstructive disease), tobacco smoking (active or passive), cancer, asthma, emphysema, fibrosis, lung allergic diseases, bronchial hyperresponsiveness and (3) lung disorders associated with immature lung epithelium e.g. occurring in pre-term babies or babies at risk for bronchopulmonary dysplasia.
Other related disorders which can be prevented and/or treated within the scope of this invention are injuries and/or diseases caused by oxidative stress, inflammation and/or aberrant responsiveness: e.g. BPD (bronchopulmonary dysplasia) in pre=
erm babies, asthma, ARDS (acute respiratory distress syndrome), atopy, allergy, fibrosis, acute lung injury etc.
The composition and medicament in accordance with the present invention may be provided to a patient by means well known in the art, i.e. orally, intranasally, subcutaneousiy, intramuscularly, intradermally, intravenously, intraarterially, parenterally or by means of catheterization.

l9 In a preferred embodiment, the invention relates to the use of a composition as described above for the preparation of a medicament for treating lung disorders or related disorders as defined above.
The present invention also relates to a method for the in vitro detection of lung disorders or other related disorders according to the invention as defined above, which method comprises the steps of:
(a) providing a sample from a body fluid of a patient, (b) contacting said sample with an antibody according to the invention as described above, to (c) optionally contacting said antibody with a secondary antibody, and (d) detecting a reaction of a molecule in said sample with said antibody.
Said body fluid can be both pulmonary {bronchoalveolar lavage (BAL), sputum, aspirate, biopsy, condensate of exhaled air, etc.) or extrapulmonary materials (serum, plasma, urine, pleural fluid, etc.).
Said method may be based on one of the techniques known to the one skilled in the art, such as for instance binding assays, especially ELISA's, RIA's, FAGS
analysis, dot blot hybridisation, Western Slots: Line immuno-assay (Lia), BIAcore real time detection system, doutie immunodiffusion technique, counterelectrophoresis technique, agglutination assay, or a mixture thereof.
2o The antibodies used in said techniques may be labelled by a tag, fluorescent marker, an enzyme, a particle or a radioactive marker. Such labels are well known to the person skilled in the art and comprise, for example, horse radish peroxidase, alkaline peroxidase, alkaline phosphatase, ~-galactosidase, fluorochromes (tike fluorescein, rhodamine, Texas Red, etc.) colloidal metals (like gold particles), biotin, digoxygenin and chemi-or bioluminescent compounds. Any detection method may be assisted by computer technology and detection methods can therefore be automated by various means.
A number of supports known in the art are suitable for use in the techniques as given above. Such supports may comprise membranes, plates, strips, wells of a 3o microtiter plate, microchips or containers. Suitable materials for such supports or materials for further coating of said supports include, but are not limited to, glass, polystyrene, polyvinyl chloride polypropylene, polyethylene, polycarbonate, dextran, nylon, amyloses, natural and modified celluloses, like nitrocellulose, poiyacrylamide, agaroses, magnetide and metals.

In a more preferred embodiment, said d~ection method is based on an ELISA
assay or a latex agglutination assay.
For example, an enzyme-linked immunosorbent assay (ELISA) can be used to 5 measure antigen concentrations, e.g. the concentration of P-SCGB in a body fluid. This ELISA method depends upon conjugation of an enzyme to an antibody, and uses the bound enzyme activity as a quantitative label. To measure said antigen concentrations, the test material containing antigen is fixed to a solid phase such as the wells of a microtiter plate, after an incubation the solid phase is washed and an enzyme-labeled io antibody, specific for said antigen, is added. Optionally, a first non-labeled antibody can be used, which is then incubated with a secondary enzyme-labeled antibody.
After washing, substrate is added, and enzyme actNity is estimated colorimetricatty, and related to antigen concentration.
As an alternative an agglutination assay rnay be used. Said agglutination assay t5 may be based on cross(linking) one of the binding partners e.g. an antibody specific for said antigen P-SCGB, to a solid support such as a bead, preferably a latex bead or a colloidal gold particle. Said beads are incubated wrth the test material containing antigen and the interactionlagglutination of said beads is measured.
Other assays than those described herein may be used, these assay formats 2o given herein are not a !imitation on the present irwention.
Said detection methods allow the diagnoss and/or monitoring of lung injuries and diseases or other related disorders such as oxidative stress related disorders and inflammation as defined above. More particularly said methods allow the diagnosis and/or monitoring of asthma.
Since the protein of the invention, P-SCGB is specifically expressed in the respiratory tract, esp. lung and trachea, and Z has a small size which facilitates its transepithelial leakage from the lung into serum, it has all the features to serve as a peripheral lung marker, easily detectable in the serum by means of an antibody. The present inventors surprisingly found that this protein was differentially expressed in 3o patients suffering from lung disorders, when compared to normal individuals. In addition, the assay of P-SCGB in urine might be used as test to detect proximal tubular dysfunction in a variety of renal diseases (e.g. cadmium nephropathy, Fanconi syndrome, diabetic nephropathy, etc.) 2t According to another embodiment, the present invention relates to a method for the detection of lung disorders or other related disorders of the invention, which method comprises the steps of:
(a) providing a sample containing nucleic acids from said patient, (b) isolating and possibly purifying nucleic acids from said sample, (c) amplifying said nucleic acids using a primer sequence as defined above, and, (d) detecting the presence of amplified DNA indicative for said lung disorders or related disorders.
According to the invention, the term "amplification' used in any method described may be performed by means of the polymerase chain reaction (PCR) and any of the primers of the invention. The amplification step is preferably done by the PCR
reaction but can also be done by any other type of nucleic acid amplification, such as 6gase chain reaction (LCR; Landgren et al., Science 241:1077, 1988; Wu and Wallace, Genomics 4:560, 1989; Barany, Proc. Natl. Acad. Sci. USA 88: 189, 1991), nucleic acid sequence based amplification (NASBA; Guatelli ei aL, Proc. Natl. Acad Sd. USA 87: 1874, 1990;
Compton, Nature 350: 91, 1991), transcription-based amplification system (TAS;
Kwoh et al., 1989), strand displacement amplificatin (SDA; Dudc, Biotechniques 9:
142, 1990;
Walker et al., Proc. Nafi. Acad. Sci. USA, 89: 392, 1992) or amplrfication by means of ass replicase (Lizardi et al., BicYTechnoiogy 6: 119? 1988; Lomeli ef al., Clin. Chem.
35: 1826, 1989) or any other suitable method to ampl'rfy nucleic acid molecules. The amplification reaction is preferably repeated between 20 and 70 times, advantageously between 25 and 45 times.
Also included in this invention are methods where the analysis of said amplified DNA as defined above and indicative for said lung disorders or related disorders (more particularly asthma) is done by direct sequencing or by micro array methods.
Also other methods can be used to analyze an amplified DNA or a mutation characteristic for said lung disorders or related disorders of the invention, including methods such as STS-PCR, countourclamped homogeneous electric field (CHEF) gel electrophoresis, restriction mapping, hybridization, Southern and Northern blotting, FISH
3o analysis, mismatch cleavage, single strand conformation polymorhism (SSCP) or any other method known in the art. The diagnostic methods of the present invention also include segregation analysis, involving PCR-based genotyping and/or haplotyping methods. The diagnostic methods according to the present invention also include methods based on direct sequencing or CAS (coupled amplification and sequencing) optionally combined with additional analytic steps as known in the art, such as ligation analysis to detect and evaluate mutations.
According to yet another embodiment, the present invention relates to a method for the detection of lung disorders or other related disorders which method comprises the s steps of:
(a) providing a sample containing nucleic acids from said patient, {b) isolating and possibly purifying nucleic acids from said sample, (c) contacting said nucleic acids with a DNA probe as defined above, and, (d) detecting a hybridization product indicative for said lung disorders or rated disorders.
Thus the detection of said lung disorders or other related disorders of the invention can also be done by means of a hybridisation reaction with any of the probes according to ~e invention. These tests generally comprise contacting the probe with the sample under hybridising conditions and detecting the presence of any duplex or triplex formation between the probe and any nucleic acid in the sample.
According to a following embodiment, the present invention relates to a method for the detection of lung disorders or other related disorders which method comprises the steps of:
{a) providing a sample from a patient, (b) contacting said sample with at ;east one of the polypeptides of as defined above, and, (c) detecting a reaction between a molecule in said sample with said polypeptide.
According to a further embodiment, the present invention relates to a method for identifying compounds for treating or preventing lung disorders or related disorders, more particularly asthma.
Such screening method may comprise the following steps of (a) contacting a compound to be tested with at least one of the polypeptides of the invention as defined above, (b) detecting the complex formed between the compound to be tested and said polypeptide, (c) alternatively, examining the diminution of complex formation between 3o said polypeptide and a receptor, caused by the addition of the compound being tested, (d) alternatively, examining the alteration in the functional activity of the polypeptide.
caused by the addition of the compound being tested, and (e) identrfying saia compound.

Said detection step in (b) is a quantitative detection step based on the affinity of said compound. A preferred example of said method is an in vitro assay based on an ELISA technique or any other method known in the art, and alkrnrs to test possible drugs which can be used in the field of lung disorders or related disorders as defined above, preferably via a high throughput method High throughput screening methods are well known in the art. Any of the well known assay formats, for example radioimmunoassays, competitive-binding assays, Western Blot analysis, antibody sandwich assays, antibody detection, ELISA
assays, fluorescence polarization, fluorescence energy transfer including fluorescence to resonance energy transfer (FRET) and homogenous time-resolved fluorescence (HTRF), fluorescence intensity, fluorescence correlation spectroscopy, sdntillation proximity assay (SPA), flash plate assays, and assays which require biotin incorporation to provide a recognition event for binding or immobilization of one or more components, etc. can be used.
The invention also relates to the compounds identifiable by the above described methods.
According to a further embodiment, the present invention also relates to an alternative compound screening method comprising the following steps of (a) providing a transgenic non-human animal as described above, in which knock-out, underexpression or overexpression of at least one of the poiypeptides of the invention results in symptoms indicative for lung disorders or related disorders, (b) administering said compound to said transgenic animal which allows said symptoms to be alleviated or cured, and (c) identifying said compound.
The invention thus relates to said non-human transgenic animals as a model system far testing potential compounds or drugs for lung disorders or related disorder, more particularly asthma. Also the compounds identifiable by this method are part of the invention.
A good example of using a transgenic animal would be the study of asthma and potential treatments of asthma using mice whose gene for P-SCGB is deleted or mutated (known as knock-out mouse or transgenic mouse). The transgenic animal may be used for the study of diseases or compounds treating diseases in the field of inflammatory and immunomodulatory lung diseases, as defined elsewhere in the patent.
Other examples describing the use of animal models as compound screening method are incorporated by the following article references, but are not considered as a limitation on the present invention (Castro C.M. et al., Lab. Invest.
80(10):1533, 2000;
Mukherjee A.B. et aL, Am. J; Kidney l7is. 32(6):1106, 199a Nizielshi S.E. et al., 126(11):2697, 1996; Aguzzi A. et al., 74(3):111, 1996; Viney J.L 4(3):461, 1994;
Metsaranfa M. et al., Ann-Med 24(2):117, 1992).
The present invention also relates to methods for treating lung disorders or related disorders (more particularly asthma) as defined above by administering to a patient a suitable nucleic acid delivery system for gene therapy. Said method may comprise administering to a patient in need a normal version of a nucleic add or gene of tt~e present invention or in the alternative switching off or krnrering the possible to overexpression of a nucleic acid or gene of the invention in a lung disorder or related disorder.
Known gene therapy protocols can consist of nucleic acid delivery systems, such as by means of expression vectors for transfection and expression of said nucleic acids as to reconstitute the function of the affected gene, or alternatively systems whereby a functional form of the affected gene or protein is delivered. Expression constructs may be administered in any biologicaly effective carrier as known in the art.
Refrovirus vectors, adenovirus vectors and adeno-associated virus vectors are exemplary nucleic acid delivery carriers for the transfer of exogenous genes in vivo, particularly into humans.
In addition to viral transfer methods, non-viral methods can also be employed, such as liposomal derived systems, poly-lysine conjugates, and artificial viral envelopes.
In clinical settings, the gene delivery systems for therapeutic use can be introduced into a patient by any of a number of methods, each of which is familiar in the art.
The present invention also relates to a pharmaceutical preparation of the gene therapy construct which can consist essentially of the gene delivery system in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery system can be produced intact from recombinant cells, e.g. retroviral vectors, the pharmaceutical preparation can comprise one or more cells which produce the gene delivery system.
3o The present invention also relates to a method for identifying compounds which selectively modulate the expression/production of a polypeptide of the invention, or alternatively which inhibit, activate or interfere with the functionality of said polypeptide of the invention, or more specific compounds which selectively inhibit, induce or interfere with one of the metabolic pathways in which said polypeptide is involved.

Such a screening method may comprise the following steps (a) providing a hose cell comprising at least part of a nucleic acid sequence of the invention in a expressible format, or transformed, transfected or infected with a vector according to the invention as described above, (b) contacting a compound to be tested with said host cell, (c) 5 monitoring the increased or decreased expression of said potypeptide caused by said compound, and, {d) identifying said compound.
The present invention further relates to a method for preparing a composition comprising a compound of the invention as detined above, comprising admixing said compound with a suitable adjuvant.
1o According to a further embodiment, the present invention relates to a diagnostic kit ~mprising an element selected from the group consisting of the nucleic adds, arrisense molecules, potypeptides, antibodies and anti-idiotypic antibodies of the invention as defined above.
Said diagnostic kit may comprise also necessary reactants and media far the diagnosis 15 according to the invention. Said diagnostic kit can be based upon a technique selected frcrn the group of techniques consisting of ELISA's, RIA's, FACS analysis, dot blot hybridisation, Western Blots, Line immuno-assay (Lia), BIAcore real time detection system, double immunadiffusion technique, countereiectrophoresis technique, agglutination assay, or additionally selected from the group consisting of .n s;tu 2o hybridisation, Northern blot hybridisation, Southern blot hybridisation, isotopic or non isotopic labelling (by immunofluorescence or biodnylated probes), genetic amplification (especially by PCR or LCR), STS-PCR, countourclamped homogeneous electric field (CHEF) gel electrophoresis, restriction mapping, FISH analysis, mismatch deavage, single strand conformation poiymorhism (SSCP) or any other method known in the art, 25 or a mixture thereat.
Said diagnostic kit thus allows the diagnosis and/or monitoring of lung injuries arid diseases or related disorders, such as inflammation- or immunomodulation-related d'sorders. More particularly, said diagnostic kit allows the diagnosis andlor monitor;ng of asth ma.
3o According to yet another embodiment, the present invention relates to a nucleic acid sequence encoding a promotor sequence comprising at least part of the sequence as defined in SEQ ID NO 3 allowing gene expression restricted to organs of the respiratory tract.

When analyzing the genomic sequence (SEG7 ID NO 3 and illustrated in Fgure 7), the inventors identified a promotor sequence upstream from the transcription start.
Said sequence presented all the features of an eukaryotic promotor and contains a characteristic sequence as given in SEO 1D NO 11. It was shown that expression of P-SCGB is very specific of the respiratory tract, especially lung and trachea.
Use of this promotor thus would allow expression of a gene restricted to lung and trachea.
This is a very useful tool, not only for expression studies in animals, but also in the prospect of gene therapy for lung disorders as set out further below.
Also included within the scope of this invention is a vector comprising said promotor sequence as defined above. Said vector can be an expression vector, as defined elsewhere in this patent application. In addition to the promotor sequence, it may be desirable to add other regulatory sequences which allow for regulation of expression.
Regulatory sequences are known to those of skill in the art, and examples include those which cause the expression of a gene to be turned on or off in response to a chemical stimulus, including the presence of a regulatory compound. Other types of regulatory elements may also be present in the vector, for example, enhancer sequences.
A host cell containing said promotor sequence or, transformed, transfected or infected with a vector comprising said sequence is also included in this invention.
Said promotor sequence of the invention as defined above can be used in the 2o generation of non-human transgenic animals. Said promotor will allow expression of genes specifically in lung tissue or other tissues of the respiratory tract.
In the transgenic animal genes may be overexpressed, underexpressed or knocked-out in said tissues by the aid of said promotor.
Said promotor sequence of the invention as defined above can also be used in nucleic acid delivery systems for gene therapy as described elsewhere in this patent application. In particular, it can be usetul in the treatment of patients with lung disorders or other related disorders (more particularly asthma), as described before in this application. In these patients, said promotor can be used for the expression of a gene restricted to the respiratory tract, especially lung and trachea. The promotor may allow 3o the expression of a normal version of a gene in the respiratory tract of said patients or alternatively, it may allow switching off or lowering a possible overexpression of a gene restricted to the respiratory tract.

2~
Another embodiment of the present invention is related to a host cell containing a nucleic acid of the invention as defined above, or transformed, transfected or infected with a vector of the invention.
According to a further embodiment, said host ceii is selected from the group comprising bacterial, fungal, insect or mammal cells.
Several documents are cited throughout this text. Each of the documents cited herein are hereby incorporated by reference, however thefe is no admission that any document cited is indeed prior art of the present invention.
Further aspects of the present invention will be described in the enclosed non-to limiting examples in reference to the following Figures.

LIST OF FIGURES
Figure 1. cDNA sequence (SEA ID NO 2) and corresponding amino acid sequence (SEA ID NO 1) of the newly identified new human branchoatveoiar protein, P-SCGB.
The peptide sequence (SEG7 ID NO 6) is given in bold.
Figure 2. Sequence alignment between P-SCGB and the consensus sequence of the family of uteroglobinslsecretoglobins.
1o Figure 3. (A) A radioactive probe was designed based on the cDNA sequence of P-SCGB (SEA 1D NO 2). The probe was used to check the expression of P-SCGB in 8 different human tissues in a Human RNA Blot (Clontech # 7760-1 ). This experiment shows that expression of P-SCGB is restricted to lung tissue.
(B) Control hybridization of the prcbe to a ubiquitos gene ((i-actin).
t5 Figure 4. Hybridization of a radioactive probe (prepared as described in F'~gure 3) to a Human Multiple Expression Array (Clontech # 7775) containing poly A+ RNA from different human tissues (+ 8 control RNAs and DNAs). This experiment confirms that the expression of P-SCGB is restricted to trachea, lung and fetal lung. The sutoradiography 2o shown in (A) must be compared to the array diagram of the different analyzed RNAs given in (B).
Figure 5. Homologuous cDNA sequence from mouse {SEA ID NO 4) and its corresponding amino acid sequence (SEQ ID NO 5).
Figure 6. Alignment of the human P-SCGB amino acid sequence (SEf~ ID NO 1 ) with the homologous mouse sequence (SEO ID NO 5).
Figure 7. Genomic sequence of the new protein P-SCGB (SEO ID NO 3). Capital letters 3o indicate bases retrieved on the cDNA given by SEO ID NO 2. Italic letters represent the promotor sequence given by SEO ID NO 1 t .
Figure 8. Schematic representation of the gene organization of P-SCGB.

EXAMPLES
EXAMPLE 1: Cloning and sequencing of a gene encoding a new human branchoalveolar protein, P-SCGB.
A new protein was identified by studying 2D-electrophoresis (2-DE) protein maps of human bronchoalveolar lavage fluids (BALFs) using a pool of BALFs from individual patients with well-defined intestitial lung diseases such as sarcoidosis, idiopathic pulmonary fibrosis and hypersensitivity pneumonitis. On these 2-DE
gels to about 600-1000 stained protein spots could be identfied by microsequence analysis and by matching with human blood plasma and the Macrophage Like Celt Line reference 2-DE maps available from the SWISS-2D PAGE database. This microsequencing gave the sequence of a short peptide (SEA 10 NO 6: APLPLDNILPFMDPLKLLLKTLGISVEH).
Frorn the peptide sequence homologies were searched in published databases.
t5 No signrficant homologies were found, indicating that this amino acid sequence does not appear in known peptide databases.
From the peptide sequence oligonudeotide primers were designed corresponding to the sequence (SEO ID NO 7: 5'-ATATCCCAGATAACTGTCATGAAGC
and SEQ ID NO 8: 5'-CCAAGTGTGATAGCGCCTCCAGCAG). Using these two primers, 2o and following a strategy known as "5' an 3' Rapid Amplification of cONA
Ends', cDNA of human origin was amplified. Amplification was done on a cDNA library from human lung (Clontech #7408), and allowed to clone two DNA fragments, which were next sequenced. This allowed to deduce the complete sequence of a cDNA (SEGl ID NO
2) expressed in human lung (Figure 1 ). This cDNA sequence is new. Comparison of the 25 sequence with all the sequences available in public databases (Genbank) shows that the sequence does not share significant homology with a previously characterized human cDNA sequence.
Analysis of the sequence and particularly translation simulations showed that this sequence is actually the coding sequence of a gene for a new protein harboring the 3o peptide sequence given by SEO ID NO 6. The amino acid sequence of the encoded protein sequence is given in SEO ID NO 1 (Figure t). Comparison of the sequence with all the sequences available in public da:abases of showed that the sequence does not share significant homology with a previously known polypeptide of human origin.

EXAMPLE 2. Structural properties of P-SCGB.
Computer simulation indicated that the new protein P-SCGB given by SEA ID
NO 1 should theoretically have a length of 93 aa, a molecular weight of 10161 Da and 5 an isoelectric point of 7.66.
However it is well known that the protein actually expressed from genes can have very drtferent characteristics, because proteins are subject to a lot of post-transtational processing such as glycosyiation, N-terminal truncation or other intracellular processing. Also, isoforms of the proteins may exist and co-exist in a living cell.
to Particular features of the protein sequence SEA ID NO 1, as well as experimental data, provide a probability that part of the sequence is cleaved after synthesis of the polypeptide and that the mature protein is shorter than 93 aa. In particular, the N-terminal part of the sequence exhibits the properties of a signal sequence, and it is likely that this part of the sequence is cleaved after or during 15 synthesis of the polypeptide. The point at which such a cleavage could occur is either between amino acids 18 and 19 or between amino acids 21 and 22, according to computer simulations.
The peptide (SEQ ID NO 6) which was isolated after 2D electrophoresis had an N-terminal sequence beginning at aminoacid 35, an apparent MW of 8500 Da and an 2o isoelectric point of 5.4. This can be due to degradation of the protein in the biological extract or to the further processing of the polypeptide (cleavage of a pre-sequence) to yield the mature P-SCGB.
This result indicates also that P-SCGB is probably decorated with haptens such as sugars or miristyl groups because the theoretical MW of a peptide beginning at as 35 25 is only 6.4 kDa and its pl 6.0, thus slightly different from observations (MW 8.5 kDa and pl 5.4). Recent data suggest that cleavage of a signal peptide occurs at or upstream from as 16 since a peptide whose N-terminal is AFLINK was identified in the 2D
electrophoresis of BASF.
3o EXAMPLE 3: Functional properties of P-SCGB indicating that it might be classified within the "uteroglobin/secretoglobin" family.
Other features of the protein suggest its classification in the family of "uteroglobinslsecretoglobins". Indeed, alignment of the P-SCGB protein with the consensus sequence of uteroglobins/secretoglobins shows signrficant similarity as illustrated in figure 2.
In the fo4lowing paragraph and related references a brief literature overview related to the 'uteroglobin/secretoglobin" family is given. For a putative biological function of P-SCGB the inventors assume properties similar to the properties of other members of the "uteroglobin/secretoglobin" family, such as anti-inflammatory, immunomodulatory, inhibition of phospholipase A2, binding of polychlorinated biphenyls (PCB's), tumor suppressor-like properties and inhibition of cancer invasiveness.
~ Uteroglobin/secretoglobin is a secreted, steroid-inducibie, progesterone binding to protein. It is also a potent immunomodulatory/antiinflammatory agent.
Indeed it is an inhibitor of phospholipase A2. (Phospholipases A2 (PLA2s; E.C.3.1.1.4) are a family of esterases that are involved in the pathogenesis of several human inflammatory diseases).
Dierynck I. et al., Mutt-Scler. t (6): 385, 1996; Mukherjee A.B. et al., DNA
Cell 8iol.
11 (3):233, 1992; Miele L et aL, Adv. Exp. Med. t3ioL 279:137, 1990; Mukherjee A.8. ef al., 32(6):1106, 1998.
~ Uteroglobinlsecretoglobin is evolutionarily conserved and secreted by the mucosal epithelial of virtually all mammals. Initially, uteroglobin/secretoglobin was identified as the major protein of rabbit uterine secretion. Counterparts of the rabbit zo uterogfobin/secretoglobin or its gene are described in rat, mouse, hamster, hare, pig, horse and human.
~ Uteroglobinlsecretoglobin is present in the blood and in other body fluids including urine, which is important for its use as a peripheral marker.
Recently, human uteroglobin/secretoglobin was shown to reverse the transformed phenotype of cancer cells and consequentiy, may have tumor suppressor-like effects.
Zhang Z. et al., Proc. Natl. Acad. Sci U S A 30;96(7):3963, 1999.
~ It has been shown that metabolites of polychlorinated biphenyls (PCBs) bind with high affinity to uteroglobin/secretoglobin, thus often called PCB-binding proteins.
Hard T. et al., Nat. Struct. Biol. 2(11):983, 1995.
~ Finally, mechanisms of uteroglobinlsecretoglobin action are likely to be even more complex as it also functions via a putative receptor-mediated pathway that has not yet been clearly defined.

EXAMPLE 4: Distribution of P-SCGB and organ specificity From the cDNA sequence SEA ID NO 2 previously determined new oligonucleotides to amplify, Gone and sequence a DNA fragment corresponding to the complete cDNA were designed. This fragment was then radioactiveiy labelled and used to probe expression of the gene in eight different human tissues using a Human RNA
Blot (Clontech #7760-1 ). The probe was hybridized to approximately 2 Ng polyA+ RNA
from the tissues. It was found that the gene is expressed in the lung, but not (or undetectable) in heart, brain, placenta, liver, skeletal muscle, kidney or pancreas (Figure to 3-A). Control hybridization of the probe to a ubiquitos gene (p-actin) is given in Figure 3-B.
In addition, to confirm the lung specifidty of the expression of P-SCGB, a Human Multiple Tissue Expression Array (Clontech #7775-1 ) was used. A radioactive probe was prepared as described above and hybridized to potyA+ RNA from 76 different human tissues and 8 different control RNAs and DNAs. In Figure 4, the autoradiography shown in (A) must be compared to the array diagram of the different analyzed RNAs given in (B).
This result shows that the protein is expressed exclusively in trachea, lung and fetal lung. It is undetectable in any other tissues tested. The tissues used for preparation of RNAs were normal, under non-pathological conditions. Cause of death was sudden death/trauma.
EXAMPLE 5: Cloning of the homologous gene from mouse.
Two oligonucleotide primers (SEO ID NO 9: 5'- CAGATA,ACTGTCATGAAGCTGGTA
and SEo ID NO 10: 5'-CCAAGTGTGATAGCGCCTCCAGCA) were designed and used to amplify cDNA from mouse lung. This allowed to amplify, clone and sequence a cDNA
fragment (given by SEO ID NO 4) which is expressed in mouse lung. The protein encoded by this sequence is given by SEQ ID NO 5. Both SEO ID NO 4 and SEO 1D
3o NO 5 are represented in Figure 5.
In Figure 6 alignment of the human P-SCGB peptide sequence (SECT ID NO 1 ) with the homologous mouse sequence (SEA VD NO 5) is given. As can be seen on this alignment, the protein encoded by the mouse cDNA is quite similar to the one encoded by the human cDNA, and thus can be considered as the mouse homologue of P-SCGB.
The identity is 79 % and the similarity is 94 %.
EXAMPLE 6: Genomic sequence of P-SCGB and chromosomal localization.
Human genome databases were searched for information relevant to the sequence of the new cDNA sequence as given by SEO ID NO 2 and revealed signrficant similarities with two large fragments of chromosome 5. This allowed to identify the genomic sequence coding for P-SCGB, as well as regulatory sequences, infom~ation for to RNA splicing, and so on. The genomic sequence given by SEO ID NO 3 is represented in Figure 7. Capital letters in Figure 7 indicate bases retrieved on the cDNA
sequence given by SEQ ID NO 2. In Fgure 8 a scheme of the gene organization of P-SCGB
is represented.
It is known that the sequence upstream from the transcription start contains a promoter and/or regulatory sequence for the expression of the gene. As it was demonstrated in the present invention that expression of P-SCGB is very specific of the respiratory tract, esp. lung and trachea, the use of this promotor andlor regulatory sequence thus will allow expression of a gene restricted to lung and trachea;
this will be a very useful tool, not only for expression studies in animals, but also in the prospect of gene therapy far lung diseases, for example. The present invention also relates to the use of nucleic acid sequence derived from those given in Figure 7 and more precisely to the sequence given in SEo ID NO 11, which presents ail features of an eukaryotic promotor.
The chromosomal assignment of P-SCGB coding information by refined by the method of Gene Radiation Hybrids. A portion of the gene for P-SCGB was amplified by PCR using 2 oligonucleotides 5'-CCTCTGGTCCCAGCTCATTTACACAG3' and 5'-TGACTATGGCCATTGCAGGCTTCTCC-3'. The Genebridge4 radiation hybrid panel was used according to Research Genetics, Inc. This panel of 93 Radiation Hybrid clones of the whole human genorne is a subset of the 199-clone panel by reported by 3o Goodfellow and Weissenbach (Walter M.A. et al., Net Genet. 7(1):22, 1994;
Gyapay G.
ef al. Hum. Mot. Genet. 5(3):339, 1996.
The obtained PCR scoring was transmitted to the "Whitehead Institute/MIT
Center for Genome Research. The locus for P-SCGB was determined as being on chromosome S, between markers D5S436 and 055470. it places 2.43 cR from W I-and 0.50 cR from WI-2452.

Claims (33)

1. An isolated nucleic acid encoding a lung marker protein or an immunologically active and/or a functional fragment of said protein selected from the group consisting of:
(a) a nucleic acid comprising or consisting of at least a functional part of the DNA
sequence as given in SEQ 10 NO 2, 3 or 4, or the complement thereof, (b) a nucleic acid which selectively hybridizes with any of the sequences as given in SEQ ID NO 2, 3, 4, 7, 8, 9 or 10, (c) a nucleic acid comprising at least a functional part of a sequence encoding a protein with an amino acid sequence which is at least 60 % homologous to the amino acid sequence as given in SEQ ID NO 1 or 5, (d) a nucleic acid encoding a protein comprising or consisting of the amino acid sequence as given in SEQ ID NO 1 or 5, (e) a nucleic acid sequence which is degenerated as a result of the genetic code to a nucleotide sequence encoding a protein as given in SEQ ID NO 1 or 5 or to a nucleic acid as defined in (a) to (d), and, (f) a nucleic acid sequence encoding a protein as defined in SEQ ID NO 1 or 5 or as defined in any one of (a) to (e) interrupted by intervening DNA sequences.

2. An isolated nucleic acid sequence according to claim 1 which is DNA, cDNA, genomic DNA, synthetic DNA or RNA wherein T is replaced by U.

3. An antisense molecule comprising a nucleic acid sequence capable of hybridizing to a nucleic acid of claim 1 or 2.

4. A probe of at least 15 contiguous nucleotides in length specifically hybridizing to a nucleic acid of claim 1 or 2.

5. A primer of at least 15 contiguous nucleotides in length specifically amplifying a nucleic acid of claim 1 or 2.

6. A vector comprising a nucleic acid sequence according to any of claims 1 to 3.

7. A vector according to claim 6 which is an expression vector wherein said nucleic acid sequence is operably linked to one or more control sequences allowing the expression in prokaryotic and/or eukaryotic host cells.

8. An isolated lung marker protein comprising one of the polypeptides selected from the group consisting of:
(a) a polypeptide as given in SEQ ID NO 1 or 5, (b) a polypeptide with an amino acid sequence which is at least 60 %
homologous to the amino acid sequence as given in SEO ID NO 1 or 5, (c) a polypeptide encoded by a nucleic acid as given in any of SEQ ID NO 2, 3 or 4, (d) a polypeptide encoded by a nucleic acid as given in any of claims 1 or 2, or a homologue or a derivative of said protein, or an immunologically active and/or functional fragment thereof.

9. A method for producing a polypeptide according to claim 8 comprising culturing a host cell comprising a vector according to claim 6 or 7, under conditions allowing the expression of said polypeptide and recovering said produced polypeptide from the culture.

10. An antibody specifically recognizing a polypeptide of claim 8 or specifically recognizing immunologically active parts or specific epitopes thereof.

11. An anti-idiotypic antibody raised against the antibody of claim 10.

12. A non-human transgenic animal transformed by a nucleic acid according to claim 1 or 2, or a vector according to claim 6 or 7.

13. A method for the production of a genetically modified non-human animal in which this modification results in overexpression, underexpression or no expression of the nucleic acids as defined in claim 1 or 2, or the polypeptides as defined in claim 8 or 9.

14. A composition comprising at least one element selected from the group consisting of a nucleic acid sequence according to any of claims 1, 2, 4 or 5; an antisense molecule according to claim 3, a polypeptide according to claim 8, an antibody according to claim 10, an anti-idiotypic antibody according to claim 11, optionally in admixture with a pharmaceutically acceptable carrier.

15. The composition according to claim 14 for use as a medicament.

16. The composition according to claim 14 for the diagnosis, prevention and/or treatment of lung disorders or ether related disorders.

17. Use of a composition according to claim 14 for the preparation of a medicament for treating lung disorders or other related disorders.

18. A method for the detection of lung disorders or other related disorders, which method comprises the steps of:

(a) providing a sample from a patient.

(b) contacting said sample with an antibody according to claim 10, (c) optionally contacting said antibody with a secondary antibody, and, (d) detecting a reaction of a molecule in said sample with said antibody.

19. A method according to claim 18 wherein said method is an enzyme linked immunosorbent assay (ELISA).

20. A method according to claim 18 wherein said method is a latex agglutination assay.

21. A method for the detection of lung disorders or other related disorders, which method comprises the steps of:

(a) providing a sample containing nucleic acids from said patient, (b) isolating and possibly purifying nucleic acids from said sample, (c) amplifying said nucleic acids using a primer sequence of claim 5, and, (d) detecting the presence of amplified DNA indicative for said lung disorders or related disorders.

22. A method for the detection of lung disorders or other related disorders which method comprises the steps of:

(a) providing a sample containing nucleic acids from said patient, (b) isolating and possibly purifying nucleic acids from said sample, (c) contacting said nucleic acids with a DNA probe of claim 4, and, (d) detecting a hybridization product indicative for said lung disorders or related disorders.

23. A method for the detection of lung disorders or other related disorders which method comprises the steps of:

(a) providing a sample from a patient, (b) contacting said sample with at least one of the polypeptides of claim 8, and, (c) detecting a reaction between a molecule in said sample and said polypeptide.

24. A method for identifying compounds for treating or preventing lung disorders or related disorders, which method comprises the steps of:

(a) contacting a compound to be tested with at least one of the polypeptides of claim 8, (b) detecting the complex formed between the compound to be tested and said polypeptide, (c) alternatively, examining the diminution of complex formation between said polypeptide and a receptor, caused by the addition of the compound being tested, (d) alternatively, examining the alteration in the functional activity of the polypeptide, caused by the addition of the compound being tested, and, (e) identifying said compound.

25. A method for identifying compounds for treating lung disorders or related disorders, which method comprises the steps of:

(a) providing a transgenic non-human animal according to claim 12 in which underexpression or overexpression of at least one of the polypeptides of claim results in symptoms indicative for lung disorders, (b) administering a compound to be tested to said transgenic animal which allows said symptoms to be alleviated or cured, and, (c) identifying said compound.

26. A method for treating lung disorders or related disorders by gene therapy which method consists of administering to a patient a suitable nucleic acid delivery system capable of reconstituting the function of the affected gene by delivering a functional form of, or switching off, or lowering a possible overexpression of a nucleic acid according to any of claims 1 or 2.

27. A method for identifying compounds which selectively modulate expression of a polypeptide of claim 8, which method comprises the steps of:

(a) providing a host cell comprising at least part of a nucleic acid as defined in any of claims 1 to 3 in an expressible format, or transformed, transfected or infected with a vector according to claim 6 or 7, (b) contacting a compound to be tested with said host cell, (c) monitoring the increased or decreased expression of said polypeptide caused by said compound, and, (d) identifying said compound.

28. A method for preparing a composition comprising a compound identifable by a method according to claims 24, 25 or 27, comprising admixing said compound with a suitable adjuvant.

29. A diagnostic kit comprising at least one element selected from the nucleic acids according to any of claims 1, 2, 4 or 5, the antisense molecules according to claim 3, the polypeptides according to claim 9, the antibodies according to claim 10, the anti-idiotypic antibody according to claim 11.

30. A nucleic acid encoding a promotor sequence comprising at least part of the sequence as defined in SEQ ID NO 3 allowing gene expression restricted to organs of the respiratory tract.

31. A vector comprising a nucleic acid sequence according to claim 30.

32. A host cell containing a nucleic acid molecule according to any of claims 1 to 3 or 30, or transformed, transfected or infected with a vector according to claim 6, 7 or 31.

33. A host cell according to claim 32 selected from the group comprising bacterial, fungal, insect or mammal cells.