KR20220122269A - Diagnostic methods for prognosis of small-cell lung cancer using CD155 and CD226 SNP - Google Patents

Abstract

The present invention relates to a method for diagnosing a prognosis of small cell lung cancer using single nucleotide polymorphisms of CD155 and CD226, and specifically, the present invention relates to a method for diagnosing or predicting a prognosis affecting the survival outcome from small cell lung cancer by using at least one single nucleotide polymorphism selected from a group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226, and to a use thereof. A technology for predicting a survival prognosis from small cell lung cancer according to the present invention can easily and accurately predict the prognosis of a patient with small cell lung cancer, can be also applied to methods for selection and evaluation of appropriate treatments, and can increase the survival rate of a small cell lung cancer patient through new targeted therapies for small cell lung cancer.

Description

Prognosis diagnosis method for small-cell lung cancer using single nucleotide polymorphisms of CD155 and CD226 {Diagnostic methods for prognosis of small-cell lung cancer using CD155 and CD226 SNP}

The present invention relates to a method for diagnosing the prognosis of small cell lung cancer using single nucleotide polymorphisms of CD155 and CD226, and specifically, diagnosing or predicting survival prognosis of small cell lung cancer using single nucleotide polymorphisms rs1058402G>A of CD155 and rs763361C>T of CD226 It relates to methods and uses thereof.

Lung cancer is known to be the leading cause of cancer-related death worldwide, and lung cancer is divided into two types: non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). Non-small cell lung cancer, for which surgery is the best treatment option at an early stage, accounts for about 85% of all lung cancers.

On the other hand, small cell lung cancer is a very aggressive carcinoma, accounting for about 15% of all lung cancers. Small cell lung cancer is predicted to be derived from neuroendocrine cells, and has a high rate of rapid tumor formation, metastasis, and recurrence. Survival and treatment methods for patients with small cell lung cancer are areas of ongoing research.

In addition, since cancer often recurs locally or metastasizes to distant tissues and organs from the original cancer tissue, it is a disease that can be effectively treated only when a treatment method is appropriately selected according to the stage. It is very important to predict

The ideal treatment for lung cancer is to detect it early and completely remove the cancer with surgery. In addition, after the onset of lung cancer, especially small cell cancer, there is very little possibility that it can be treated through surgical operation when it is detected. Current clinical diagnostic methods cannot confirm the prognosis of early small cell lung cancer with sufficient accuracy to instruct more aggressive therapy for patients with a high risk of relapse.

Therefore, it is necessary to apply a treatment method by determining the type of chemotherapy according to the stage and characteristics, and it is necessary to determine the prognosis of small cell lung cancer patients accurately and quickly so that it is easy to determine whether to use additional necessary treatment methods. There is a need for diagnostic technology.

Republic of Korea Patent Registration 10-2195593

Accordingly, the present inventors can use the rs1058402G>A single nucleotide polymorphism of CD155 or the rs763361C>T polymorphism of CD226 as a new diagnostic marker related to the development and progression of small cell lung cancer (SCLC), and specific genotypes of these polymorphisms and small cell lung cancer The present invention was completed by identifying the relevance to the patient's survival prognosis.

Accordingly, it is an object of the present invention to provide a marker composition for prognosis and prediction of small cell lung cancer comprising at least one single nucleotide polymorphism selected from the group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226 will do

Another object of the present invention is to predict the survival prognosis of a patient with small cell lung cancer, including an agent capable of detecting one or more single nucleotide polymorphisms selected from the group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226 To provide a composition for

Another object of the present invention is to provide a kit for predicting survival prognosis of small cell lung cancer patients comprising the composition of the present invention.

Another object of the present invention is to provide a microarray for predicting survival prognosis of a patient with small cell lung cancer comprising the composition of the present invention.

Another object of the present invention is to provide information for predicting survival prognosis of patients with small cell lung cancer, including identifying the rs1058402G>A single nucleotide polymorphism of CD155 and the rs763361C>T single nucleotide polymorphism of CD226 with respect to the nucleic acid extracted from the sample. It is to provide a way to provide.

In order to achieve the above object of the present invention, the present invention provides a prognosis diagnosis of small cell lung cancer and A composition for a marker for prediction is provided.

In one embodiment of the present invention, the rs1058402G>A single nucleotide polymorphism of CD155 is located at the 250th base in the nucleotide sequence consisting of SEQ ID NO: 1, and the rs763361C>T single nucleotide polymorphism of CD226 consists of SEQ ID NO: 2 It may be located at the 500th base in the base sequence.

In one embodiment of the present invention, the small cell lung cancer may be treated using chemotherapy.

In addition, the present invention provides an agent capable of detecting one or more single nucleotide polymorphisms selected from the group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226, for predicting survival prognosis of patients with small cell lung cancer A composition is provided.

In one embodiment of the present invention, the agent is a polynucleotide consisting of 10 to 100 consecutive bases comprising rs1058402G>A single nucleotide polymorphism of CD155 or rs763361C>T single nucleotide polymorphism of CD226 or a polynucleotide complementary thereto It may be a primer or probe capable of amplifying.

In one embodiment of the present invention, the rs1058402G>A single nucleotide polymorphism of CD155 is located at the 250th base in the nucleotide sequence consisting of SEQ ID NO: 1, and the rs763361C>T single nucleotide polymorphism of CD226 consists of SEQ ID NO: 2 It may be located at the 500th base in the base sequence.

In one embodiment of the present invention, the agent comprises a primer pair consisting of nucleotide sequences of SEQ ID NOs: 3 and 4 capable of detecting the rs1058402G>A single nucleotide polymorphism of CD155; Alternatively, it may be a primer pair consisting of the nucleotide sequences of SEQ ID NOs: 5 and 6 capable of detecting the rs763361C>T single nucleotide polymorphism of CD226.

In one embodiment of the present invention, the rs1058402G>A single nucleotide polymorphism of CD155 or the rs763361C>T single nucleotide polymorphism of CD226 is a chemotherapy response to an anticancer agent, overall survival (OS), or progression-free It may be related to progression-free survival (PFS).

The present invention also provides a kit for predicting survival prognosis of small cell lung cancer patients comprising the composition of the present invention.

The present invention also provides a microarray for predicting survival prognosis of a patient with small cell lung cancer comprising the composition of the present invention.

Furthermore, the present invention provides information for predicting survival prognosis of patients with small cell lung cancer, including confirming the rs1058402G>A single nucleotide polymorphism of CD155 and the rs763361C>T single nucleotide polymorphism of CD226 with respect to the nucleic acid extracted from the sample. provide a way

In one embodiment of the present invention, when the genotype of the rs1058402G>A polymorphism of CD155 is GG, it can be determined as a group having a higher survival prognosis than in the case of GA or AA.

In one embodiment of the present invention, when the genotype of the rs763361C>T polymorphism of CD226 is CT or TT, it can be determined as a group having a higher survival prognosis than CC.

In one embodiment of the present invention, the GA or AA genotype of the rs1058402G>A polymorphism of CD155; Or, when the CC genotype of the rs763361C>T polymorphism of CD226 is 0, the chemotherapy response to the anticancer agent is 80.4%, and in the case of one, the chemotherapy response to the anticancer agent is 71.0%, and in the case of two, the chemotherapy response to the anticancer agent The therapy response may be 61.3%.

In one embodiment of the present invention, when alanine (A), the 67th amino acid of CD155, is mutated to threonine (T) in the rs1058402G>A polymorphism of CD155, chemotherapy response to an anticancer agent, overall survival rate (Overall survival: OS) or progression-free survival (PFS) may be determined to be low.

In one embodiment of the present invention, when alanine (A), the 67th amino acid of CD155, is mutated to threonine (T) in the rs1058402G>A polymorphism of CD155, the binding force between CD155 and CD226 is reduced, thereby reducing T cell activity or NK cell activity. activity may be reduced.

The technology for predicting the survival prognosis of small cell lung cancer according to the present invention can rapidly and accurately predict the prognosis or survival prognosis of the patient's anticancer drug treatment for a patient who has been diagnosed with small cell lung cancer and has received anticancer drug treatment. It can also be applied to the method, and furthermore, the survival rate of small cell lung cancer patients can be increased through a new targeted treatment for small cell lung cancer.

1 shows a schematic diagram of a process for selecting a patient group in an embodiment of the present invention.
2 shows a Kaplan-Meier plot for overall survival according to polymorphism in an embodiment of the present invention, where A is CD155 rs1058402, B is CD226 rs763361, (C) is the association with overall survival for a combination of bad genotypes. is shown. P values were calculated using a multivariate Cox proportional hazards model.
Figure 3 shows the structural modeling analysis results for the CD226/CD155 (wild-type or A67T mutant) complex, where A is mCD226-ecto (grey) and hCD155-Domain1 (orange) aligned with the structural model of hCD155-T67 (green). The 3D structure of the structure (PDB ID: 6ISC) is shown, B and C are the structural models for the interaction between CD226-Domain1-N116 and CD155-G73, S74, A67/T67, and D is CD226-Domain2- A structural model for the interaction between E185 and CD155- wild-type/mutant model G70 is shown. Relevant amino acid residues are indicated by bars (nitrogen atom: blue, oxygen atom: red). The interatomic distance (angstroms) is indicated by the black dotted line, and the estimated polarity junction is indicated by the red line. All images of CD226/CD155 (wild-type versus A67T mutant) were formed using PyMOL (https://pymol.org/2/).

The present invention has identified that single nucleotide polymorphisms of CD155 and CD226 can be used as biomarkers for prognostic diagnosis and prediction of small cell lung cancer. Specifically, the present invention provides rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T of CD226 It is characterized by providing a composition for a marker for the diagnosis and prediction of prognosis of small cell lung cancer comprising one or more single nucleotide polymorphisms selected from the group consisting of single nucleotide polymorphisms.

CD155 can be represented by SEQ ID NO: 1.

SEQ ID NO: 1: ACACCCCACG GTCCACCGGG GATCCAGGGC CCCAGCGTGC AAGTCGCGCC AGGTGCCCGG

GCTCCTGGAG CCCCTCCCTA TCTAGTCCAA GAACGCCCCG GGTCTGACAC CTTCTCTTCG GTTCTCCGCA GGGGACGTCG TCGTGCAGGC GCCCACCCAG GTGCCCGGCT TCTTGGGCGA CTCCGTGACG CTGCCCTGCT ACCTACAGGT GCCCAACATG GAGGTGACGC ATGTGTCACA GCTGACTTG G CGCGGCATGG TGAATCTGGC AGCATGGCCG TCTTCCACCA AACGCAGGGC CCCAGCTATT CGGAGTCCAA ACGGCTGGAA TTCGTGGCAG CCAGACTGGG CGCGGAGCTG CGGAATGCCT CGCTGAGGAT GTTCGGGTTG CGCGTAGAGG ATGAAGGCAA CTACACCTGC CTGTTCGTCA CGTTCCCGCA GGGCAGCAGG AGCGTGGATA TCTGGCTCCG AGTGCTTGGT GAGCAGGGGG TTTTGGGGAG

CD226 can be represented by SEQ ID NO:2.

SEQ ID NO: 2: CATCTTGCCA ACACAAGAGG AAAGCCTGCA TGAGAGTGAG GCCAAGACCA GGGAAATGGA

TCCAACACTC TGGAAAGGGA TTCAGAAGCC AGTTTATGCC CATACTTAAT TCTAGACACA ACATTTAAGC CCTGGTAAAT AGCCCTTGCC CAAAGCTTAA TCTCCCCTGG ATCATTCTGT TATATGATAC AGTAAGTTTT CTTTTGTATA TAAACCAGTT AGAGTCAGGT TTTCTGAAAC AGTTCTATGA AAAATGATTT TAGGTAATGA AGTATTTTTC CTATCAAAGT AACTCAATTC AGACAACTAG TATCTAAGGT AGACCTTGGG TAGTGGAAAA AAATTGCATA AAGATCCATG CATGAGTACA TAAGAGTCAT TACTAATGCA CTCATGTCAA GAATAAGCTT AAACTCTAGT CTTTGGTCTG CGAGAGAAGG TTGGATAGTT GACATAAATA TCCTCTCTTG TATCATCCAT GGATTGATTG GTAGGTTGA C GGTAGAGATG GGACTTCTAT AGTTATTGGG TGCCTAGAAA GACAAACAGC AGAGAGTGTC AATAATTCAC TGCATTTCAA CAACTAATGA AGACATTCAA AACATACCTC TCATAAATGC AGGCATGATA TGGGACAAGT TTCCATTATA ATTAATTTGA TAAAACATTC TCATTGTATT GAACTTCTGT TGGAAATCAT ATTCTTGACT GAGATGCAAC AGAAATCTAG TCTAGATTAA TACTTGCTGT ATTGTGCTAT GAATTACCTG TTTCAATGTA GAACTCCCTC TCCCAATGTT AAGCTATTTT GATAAGAACA ATTACTATCA TTTTTATATG GAAATTCTAA CACAGTGTCT GAACTTTAAA AAATAAGCAA CTTAACATAT AAAAAGTTAA CAAAACTGAC ATCAAGATGA TATTGTATAA TCATTAATCT TCTCTACCGT AATCAAGTCA CTTTGACCA C ATATGTATGT ATGTAACAGA AAATATGCAG

While the present inventors were researching to discover biomarkers that can rapidly and accurately diagnose and predict the prognosis of small cell lung cancer, immune checkpoint genes and association with small cell lung cancer as candidate markers related to the development and progression of small cell lung cancer (SCLC) Selected genes with this gene and analyzed whether the single nucleotide polymorphism present in the genes was correlated with the clinical results of patients who received chemotherapy for small cell lung cancer. It was confirmed that the rs763361C>T single nucleotide polymorphism of CD226 was significantly related to the chemotherapy response to small cell lung cancer and the survival prognosis of the patient.

Meanwhile, CD155, also called PVR or Necl-5, encodes a transmembrane glycoprotein belonging to the immunoglobulin superfamily. CD155 was initially known to act on cell adhesion and migration, but recently it has also been reported to have relevance in immunology and oncology.

CD226 also encodes for glycoproteins expressed on the surface of natural killer (NK) cells, T cells, subsets of B cells and monocytes and plays an important role in their activation and inhibition. CD155 present in antigen-presenting cells or tumor cells binds to CD226 present in T cells and NK cells, and the binding of CD155 and CD226 acts as a co-stimulator to activate T cells and NK cells.

In this regard, the present inventors studied the association between the rs1058402G>A polymorphism in CD155 and the rs763361C>T polymorphism in CD226 with small cell lung cancer. As a result, the genotype of the rs1058402G>A polymorphism in CD155 is GG. case, the survival prognosis was higher than that of GA or AA, and when the genotype of the rs763361C>T polymorphism of CD226 was CT or TT, the survival prognosis was higher than that of CC.

Therefore, it was found that the survival prognosis of small cell lung cancer patients can be predicted or diagnosed through genotyping of these single nucleotide polymorphisms.

On the other hand, the rs1058402G>A polymorphism present in CD155 is a miss sense mutation. rs1058402G>A has a mutant form in which alanine (A) at position 67 of the amino acid codon of CD155 is substituted with threonine (T). In one embodiment of the present invention, it was confirmed that the distance to S74/G73 of CD155 was shortened when alanine at codon 67 of CD155 was changed to threonine through the 3-D structural model in an embodiment of the present invention. It was confirmed that the shortened distance formed a new hydrogen bond between T67 and S74 of CD155, and this change induced an extension of the distance between S74 of CD155 and N116 of CD226 and loss of hydrogen bond.

In addition, the structural alteration due to rs1058402G>A(A67T) was shown to increase the bonding distance between G70 of CD155 and E185 of CD226, leading to loss of hydrogen bonds, and S74/G70 of CD155 in the interaction between CD155 and CD226. and N116/E185 of CD226 were confirmed to be important binding sites.

In an example of the present invention, it was found that the CD155 rs1058402G>A(A67T) polymorphism was significantly associated with worse chemotherapy response and overall survival in patients with small cell lung cancer, and 3D structural model analysis showed that rs1058402G>A(A67T) was It was found that by increasing the distance of the binding site between CD155 and CD226, the binding force between CD155 and CD226 was weakened. It was found that the treatment effect and survival rate were negatively affected in lung cancer patients. In the present invention, the amino acid sequence of CD155 is shown in SEQ ID NO: 7, and the CD155 rs1058402G>A(A67T) mutation position corresponds to the 67th amino acid of SEQ ID NO: 7.

In addition, the CD226 rs763361C>T polymorphism was shown to be associated with better chemotherapy response and overall survival in patients with small cell lung cancer, where rs763361C>T is located in exon 7, encoding the cytoplasmic tail of CD226, 2 It has two phosphorylation sites and is a non-synonymous mutation. In the rs763361 polymorphism, the substitution of the C base with T leads to the substitution of serine for glycine at amino acid codon 307. It was found that the Gly307Ser substitution affects CD226 expression by altering phosphorylation or by disrupting the exon splicing silencer sequence to alter RNA splicing.

In the present invention, the amino acid sequence of CD226 is shown in SEQ ID NO: 8, and the mutation position of rs763361C>T (Gly307Ser) corresponds to the 307th amino acid of SEQ ID NO: 8.

Therefore, in the present invention, polymorphisms in the immune checkpoint genes CD155 and CD226 are related to the survival outcome of small cell lung cancer patients who have received chemotherapy, and mutations in CD155 and CD226 are more important than immunotherapy alone or chemotherapy and immunity than chemotherapy alone. It was found that it can have a greater influence on predicting clinical outcomes according to the combination treatment of therapy.

Furthermore, in the present invention, it was confirmed that mutants and mutants of the immune checkpoint gene, CD155 rs1058402G>A(A67T) and CD226 rs763361C>T(G307S), are related to chemotherapy response and overall survival, in particular, CD155 rs1058402G>A(A67T). ) could affect the interaction and binding force between CD155 and CD226.

Therefore, the present invention provides a marker composition for diagnosing and predicting the prognosis of small cell lung cancer, comprising one or more single nucleotide polymorphisms selected from the group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226. can

In the present invention, the rs1058402G>A single nucleotide polymorphism of CD155 is located at the 250th nucleotide in the nucleotide sequence consisting of SEQ ID NO: 1, and the rs763361C>T single nucleotide polymorphism of CD226 is located at the 500th nucleotide in the nucleotide sequence consisting of SEQ ID NO: 2 do.

In addition, the present invention provides an agent capable of detecting one or more single nucleotide polymorphisms selected from the group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226, for predicting survival prognosis of patients with small cell lung cancer A composition is provided.

The agent may be a primer or probe capable of amplifying a polynucleotide consisting of 10 to 100 consecutive bases containing rs1058402G>A single nucleotide polymorphism of CD155 or rs763361C>T polymorphism of CD226 or a complementary polynucleotide thereof have.

Preferably, the agent is a primer pair consisting of nucleotide sequences of SEQ ID NOs: 3 and 4 capable of detecting rs1058402G>A single nucleotide polymorphism of CD155 or rs763361C>T of SEQ ID NOs: 5 and 6 capable of detecting rs763361C>T single nucleotide polymorphism of CD226 It may be a primer pair consisting of a nucleotide sequence.

In addition, according to the present invention, the rs1058402G>A single nucleotide polymorphism of CD155 or the rs763361C>T single nucleotide polymorphism of CD226 according to the present invention is a chemotherapy response, overall survival (OS) or progression-free survival rate (Progression-) free survival (PFS)).

In the present invention, the "prognosis" refers to the progress and cure of diseases such as the onset, recurrence, metastatic spread, and the possibility of lung cancer-induced death or progression, including drug resistance, of a disease such as lung cancer. For the purposes of the present invention, the prognosis means the risk of developing lung cancer, preferably small cell lung cancer, and the prognosis for survival after the onset.

"Prediction" means determining whether a patient is likely to develop lung cancer, preferably small cell lung cancer, and responding favorably or unfavorably to a treatment, such as chemotherapy or radiation, to treat the patient, e.g., a specific therapeutic agent. , and/or surgical removal of the primary tumor, and/or survival and/or likelihood of survival after treatment with chemotherapy for a specified period of time without cancer recurrence.

The prediction method of the present invention is a patient with a high risk of developing small cell lung cancer for any specific patient, and through special and appropriate management, the onset time is delayed or the onset is not delayed, or the treatment is performed by selecting the most appropriate treatment method for the small cell lung cancer patient. It can be used clinically to make decisions. The predictive method of the present invention determines whether a patient responds favorably to a treatment regimen, such as a prescribed treatment regimen, including, for example, administration of a given therapeutic agent or combination, surgical intervention, chemotherapy, etc. It is possible to predict whether or not survival is possible.

"Genetic polymorphism" refers to the occurrence of a genetic mutation with a frequency of at least 1% or more in a population. The insertion, deletion, or substitution of one nucleotide in DNA is called single nucleotide polymorphism (SNP).

The term "polymorphism" refers to the placement in the sequence of a gene that varies within a population. Polymorphisms consist of different “alleles”. The placement of such polymorphisms can be identified by their position in the gene and the different amino acids or bases found therein. These amino acid variations are the result of two different alleles, two possible variant bases. Because the genotype consists of two different distinct alleles, any of several possible variants can be observed in any one individual. Individual polymorphisms are also known to those skilled in the art and are used, for example, in the Single Nucleotide Polymorphism Database (dbSNP) of Nucleotide Sequence Variation available on the NCBI website. Identifier ("reference SNP", "refSNP" or "rs#").

"Single nucleotide polymorphism (SNP)" is the diversity of DNA sequences that occurs when a single nucleotide (A, T, C, or G) in the genome differs between members of a species or between pairs of chromosomes in an individual. means For example, when three DNA fragments from different individuals (eg, AAGT[A/A]AG, AAGT[A/G]AG, AAGT[G/G]AG) contain differences in a single base, Called two alleles (A or G), in general almost all SNPs have two alleles. Within a population, SNPs can be assigned a minor allele frequency (MAF; the lowest allele frequency at a locus found in a particular population). A single base can be changed (replaced), removed (deleted), or added (inserted) into the polynucleotide sequence. SNPs can cause changes in the translation frame.

The term “genotype” refers to a specific allele of a particular gene in a cell or tissue sample.

"Allele" or "allele" means one of two or more alternative forms of a gene that occupy the same chromosomal locus.

"Diagnosing" means identifying the presence or character of a pathological condition. Among them, the present invention particularly includes predicting recurrence, progression, metastasis, and the like of small cell lung cancer. Therefore, an indicator that can accurately predict the recurrence and progression of small cell lung cancer is very important, and a factor that can predict the response to treatment while supplementing clinical indicators such as the degree of differentiation and stage of the tissue is needed. Since the rs763361C>T SNP of SNP or CD226 functions as such an indicator, it can be used as a diagnostic factor for small cell lung cancer. That is, measurement of polymorphism of these genes can be used as a useful indicator (diagnostic marker) for predicting the degree of differentiation, stage, and progression of small cell lung cancer.

"Diagnostic marker or diagnostic marker" is a substance that can differentiate and diagnose cells with small cell lung cancer from normal cells, and a polypeptide or nucleic acid (e.g., : mRNA, etc.), lipids, glycolipids, glycoproteins, and organic biomolecules such as sugars (monosaccharides, disaccharides, polysaccharides, etc.).

"Marker for predicting survival prognosis of patients with small cell lung cancer" means a marker having a polymorphism that can predict the risk of small cell lung cancer, whether or not the onset of small cell lung cancer is cured, or the course, and preferably refers to the nucleotides described above. . In addition, the patient refers to a patient for determining the risk of developing small cell lung cancer or a patient who has undergone chemotherapy for small cell lung cancer.

"Subject" or "patient" means any single individual in need of treatment, including humans, cattle, dogs, guinea pigs, rabbits, chickens, insects, and the like. Also included in the subject are any subjects who participated in a clinical study trial that do not show any clinical manifestations of any disease, or subjects who participated in epidemiological studies or subjects used as controls.

"Tissue or cell sample" means a collection of similar cells obtained from the tissue of a subject or patient. Sources of tissue or cell samples may include solid tissue from fresh, frozen and/or preserved organ or tissue samples or biopsies or aspirates; blood or any blood component; The cells may be at any time of conception or development in a subject. A tissue sample may also be a primary or cultured cell or cell line.

"Nucleic acid" is meant to include any DNA or RNA, eg, chromosomal, mitochondrial, viral and/or bacterial nucleic acids present in a tissue sample. It includes one or both strands of a double-stranded nucleic acid molecule, and includes any fragment or portion of an intact nucleic acid molecule.

By “gene” is meant any nucleic acid sequence or portion thereof that has a functional role in protein coding or transcription or in the regulation of other gene expression. A gene may consist of any nucleic acid encoding a functional protein or only a portion of a nucleic acid encoding or expressing a protein. Nucleic acid sequences may include gene abnormalities within exons, introns, initiation or termination regions, promoter sequences, other regulatory sequences, or unique sequences adjacent to the gene.

A "primer" is an oligonucleotide sequence that hybridizes to a complementary RNA or DNA target polynucleotide and serves as a starting point for the stepwise synthesis of polynucleotides from mononucleotides, for example, by the action of nucleotidyl transferases occurring in the polymerase chain reaction. means

"Protein" also includes fragments, analogs and derivatives of proteins that retain essentially the same biological activity or function as a reference protein.

"Treatment" means an approach for obtaining beneficial or desirable clinical results. For the purposes of the present invention, beneficial or desirable clinical outcomes include, but are not limited to, alleviation of symptoms, reduction of the extent of the disease, stabilization (ie, not worsening) of the disease state, delay or reduction in the rate of disease progression, disease state improvement or temporary alleviation and alleviation (partial or total), detectable or undetectable. "Treatment" may also mean increasing survival as compared to the expected survival rate if not receiving treatment. Treatment refers to both therapeutic treatment and prophylactic or prophylactic methods. Such treatments include the treatment required for the disorder being prevented as well as the disorder that has already occurred. "Palliating" a disease means that the extent and/or undesirable clinical signs of the disease state are reduced and/or the time course of progression is delayed or prolonged, compared to no treatment. means to lose

In addition, the present invention can provide a kit for predicting the survival prognosis of a small cell lung cancer patient comprising the composition according to the present invention, and further, it is possible to provide a microarray for predicting the survival prognosis of a small cell lung cancer patient comprising the composition according to the present invention. have.

The kit of the present invention can be used for predicting small cell lung cancer survival prognosis by identifying polymorphic sites of rs1058402G>A of CD155 or rs763361C>T of CD226, which are markers for predicting small cell lung cancer survival prognosis. The kit for predicting the survival prognosis of small cell lung cancer of the present invention may include polynucleotides, primers or probes for confirming the polymorphism, as well as one or more other component compositions, solutions or devices suitable for the analysis method.

In addition, the kit of the present invention may be a kit including essential elements necessary for performing PCR. The PCR kit contains, in addition to polynucleotides, primers or probes specific for the SNP, a test tube or other suitable container, reaction buffer (with varying pH and magnesium concentrations), deoxynucleotides (dNTPs), Taq-polymerase and reverse transcriptase. enzymes such as DNase, RNAse inhibitors, DEPC-water and sterile water, and the like.

In addition, the microarray of the present invention may be composed of a conventional microarray except for containing the polynucleotide, primer or probe of the present invention.

Hybridization of nucleic acids on microarrays and detection of hybridization results are well known in the art. The detection is, for example, labeling the nucleic acid sample with a labeling material capable of generating a detectable signal including a fluorescent material, for example, a material such as Cy3 and Cy5, and then hybridizing on a microarray and the labeling material. A hybridization result can be detected by detecting a signal generated from

The microarray method can simultaneously study the RNA expression of thousands or even tens of thousands of genes within a tumor, allowing more effective insight into the molecular basis of human disease.

It can also be used to evaluate gene expression patterns in tumor classification, clinical outcomes, and response to chemotherapy.

Furthermore, the present invention can provide a method for providing information for predicting the survival prognosis of a patient with small cell lung cancer, wherein the method preferably includes rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C> of CD226 with respect to the nucleic acid extracted from the sample. and identifying the T single nucleotide polymorphism.

In this case, when the genotype of the rs1058402G>A polymorphism of CD155 is GG, it can be determined as a group having a higher survival prognosis compared to the case of GA or AA, and when the genotype of the rs763361C>T polymorphism of CD226 is CT or TT , it can be determined as a group with a higher survival prognosis than CC.

In one embodiment of the present invention, the GA or AA genotype of rs1058402G>A polymorphism of CD155 with poor prognosis for the genotypes for the two polymorphisms; Alternatively, the number of CC genotypes of the rs763361C>T polymorphism of CD226 was measured and the relationship between the number of the bad genotypes and the chemotherapy response to the anticancer agent was analyzed. As a result, the number of the bad genotypes was 0. The chemotherapy response to chemotherapy was 80.4%, and in the case of one patient, the chemotherapy response to the anticancer agent was 71.0%, and in the case of two patients, the chemotherapy response to the anticancer agent was 61.3%.

In addition, through mutation and protein structure analysis of the rs1058402G>A polymorphism of CD155, mutations from alanine (A), the 67th amino acid of CD155, to threonine (T) in the rs1058402G>A polymorphism of CD155, chemotherapy response to anticancer drugs ( chemotherapy response), overall survival (OS), or progression-free survival (PFS) can be judged to be low, and the binding force between CD155 and CD226 is reduced, resulting in a decrease in T cell activity or NK cell activity do.

On the other hand, in the method of providing information for predicting the survival prognosis of the small cell lung cancer patient according to the present invention, the method of confirming the polymorphism is a method of confirming the genotype of the rs1058402G>A of CD155 and rs763361C>T of the CD226 polymorphism. The genotype can be confirmed by sequencing analysis, sequencing analysis using an automatic sequencing analyzer, pyrosequencing, hybridization by microarray, PCR-RELP method (restriction fragment length polymorphism), PCR-SSCP method (single strand conformation polymorphism), PCR-SSO method (specific sequence oligonucleotide), ASO (allele specific oligonucleotide) hybridization method combining PCR-SSO method and dot hybridization method, TaqMan-PCR method, MALDI-TOF/MS method, RCA method (rolling circle amplification), HRM (high resolution melting) method, primer extension method, Southern blot hybridization method, it can be carried out by a known method such as a dot hybridization method.

In addition, the results of the SNP polymorphism can be statistically processed using statistical analysis methods commonly used in the art, for example, Student's t-test, chi-square test (Chi-) square test), linear regression line analysis, and multiple logistic regression analysis, such as continuous variables, categorical variables, odds ratio, and 95 It can be analyzed using variables such as % confidence interval.

In addition, the diagnostic method relates to examining the expression characteristics of specific markers according to small cell lung cancer, and the method disclosed herein is convenient and useful for obtaining useful data and information when evaluating an appropriate or effective therapy for treating a patient with small cell lung cancer. , it will be possible to provide an efficient and cost-effective means.

Nucleic acids from small cell lung cancer patients can be obtained from samples such as tissues, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid or urine obtained from these patients, and the nucleic acid sample is DNA, mRNA, or synthesized from mRNA. contains cDNA.

The nucleic acid of the small cell lung cancer patient can be performed by conventional methods and separation methods such as phenol/chloroform extraction and protease K treatment, and can also be obtained by amplifying the target nucleic acid through PCR and purifying it.

In addition, the present invention can also provide information for determining the amount and administration method of an anticancer agent necessary to prevent recurrence or metastasis after chemotherapy treatment for small cell lung cancer.

That is, the present invention confirmed the rs1058402G>A polymorphism of CD155 and the rs763361C>T polymorphism of CD226 from a biological sample isolated from a patient with small cell lung cancer, and the type, amount, and concentration of the anticancer agent to be additionally administered depending on the case of poor prognosis or otherwise Anticancer drug administration methods, such as, may be applied differently to suit the type of cancer.

Hereinafter, the components and technical features of the present invention will be described in more detail through the following examples. However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited by the examples.

<Test materials and methods>

1. Selection of research subjects

This study was conducted as an observational retrospective study. From March 2001 to November 2017, 261 small cell lung cancer (SCLC) patients who were clinically diagnosed at Daegu Kyungpook National University Hospital (KNUH) were included. The patient selection process is shown in FIG. 1 . All patients received platinum doublet chemotherapy as the primary treatment and received at least 2 cycles of chemotherapy. In addition, patients with limited-stage SCLC who received chemotherapy in conjunction with radiation therapy were excluded from this study to avoid confounding effects of radiation on chemotherapy response. Patients who received radiotherapy sequentially after 2 cycles of chemotherapy were included. In this case, the response after 2 cycles of chemotherapy was considered as the best response to chemotherapy. Two chemotherapy regimens were applied to the patients, one of which was cisplatin 60 mg/m ² on day 1 and etoposide 100 mg/m ² every 3 weeks on days 1, 2 and 3 administered. In another method, cisplatin 60 mg/m ² was administered on the 1st day, and irinotecan 60 mg//m ² was administered every 4 weeks on the 1st, 8th, and 15th days. The decision to maintain treatment was based on the patient's disease progression, major toxicity, or the patient's or physician's decision. Response to chemotherapy was assessed using the response criteria for solid tumors. Chemotherapy response categories were divided into responders and non-responders. Patients with complete or partial response to responders were considered best responders, and patients with stable disease or progressive disease were considered non-responders.

This study was conducted with the approval of the institutional review committee of the Korea Advanced Institute of Science and Technology. Blood samples for genotyping were provided and used by the National Institutes of Health, National Biobank, supported by the Ministry of Health and Welfare. All blood samples were collected prior to first-line chemotherapy, and all subjects were 18 years of age or older, and informed consent was obtained prior to chemotherapy.

2. Polymorphism Selection and Genotyping

38 genes involved in immune checkpoint pathways were selected by searching public databases and related literature. The public SNP database (http://www.ncbi.nlm.nih.gov/SNP) and RegulomeDB (http://www.regulomedb.org/) were used to collect polymorphisms of immune checkpoint genes. After excluding minor allele frequencies of ≤0.05 based on HapMap JPT data, 216 potential functionalities were identified using the FuncPred utility for functional SNP prediction on the SNPinfo web server (https://snpinfo.niehs.nih.gov/). SNPs were collected. For LD-tagged SNP selection, 123 SNPs were selected using the TagSNP utility, excluding those with linkage disequilibrium (LD) coefficient r ² greater than or equal to 0.8. Of the 123 SNPs, 27 with a call rate of less than 95% or a P value of less than 0.05 for Hardy-Weinberg equilibrium (HWE) were excluded from the analysis. 96 SNPs from 33 immune checkpoint genes were selected for analysis for association and response studies. In addition, genotyping was performed using the Sequenom MassARRAY iPLEX Platform (Agena Bioscience, San Diego, USA). All genotyping analyzes were performed blindly with respect to the patient's condition, and all methods were performed according to the relevant guidelines and regulations.

3. Statistical processing

HWE was analyzed using the goodness-of-fit χ2 test with 1 degree of freedom. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression analysis for comparisons between clinical variables or genotype and chemotherapy response. Overall survival (OS) was analyzed from first chemotherapy treatment to death or last follow-up. Progression-free survival (PFS) was analyzed from the day of first-line chemotherapy until disease progression or death from any cause. The Kaplan-Meier method was used to estimate survival outcomes, and the log-rank test was used to compare OS in different groups or genotypes. Hazard ratios (HR) and 95% CI were estimated using a multivariate Cox proportional hazards model. Adjustment factors were age, sex, smoking status, stage, ECOG (Eastern Cooperative Oncology Group) performance status, weight loss, neuron specific enolase (NSE) level, first-line chemotherapy, second-line chemotherapy, and radiation for primary tumor. . P values less than 0.05 were considered statistically significant, and statistical analysis was performed using Statistical Analysis System Version 9.4 for Windows (SAS Institute, Cary, NC, USA).

4. Structural Modeling for the CD155 A67T Mutation

To confirm the effect on the mutation of alanine (A) to threonine (T) at codon 67 on the rs1058402G>A polymorphism of CD155, the mutant structure of CD155 (A67T) was used to determine wild-type CD155 using MODELLER v9.12. The model was compared with the structure (PDB: 6ISC). At least 10 structural violations were identified by model/fine loop (loop modeling protocol DOPE) using UCSF Chimera v1.14, the structure showed no violations and the lowest energy was chosen as the model. All images of CD226/CD155 (wild-type VS A67T mutant) were generated using PyMOL (https://pymol.org/2/).

<Example 1> Patient characteristics and clinical predictors

The results of the chemotherapy response and overall survival (OS) according to the clinical characteristics of the patients are shown in Table 1 below. The overall response rate was found to be 72.8% (95% CI = 67.4-78.2%). The median survival time (MST) was found to be 10.4 months (95% CI = 9.1-11.1 months). The response rate was higher with the irinotecan/cisplatin regimen than with the etoposide/cisplatin regimen (78.7% vs 67.2%, P = 0.04), and the OS did not differ according to the regimen (MST, 10.0 months vs 10.7 months, P = 0.88, Table 1). OS, age, stage, ECOG performance status, weight loss, NSE level, second-line chemotherapy and tumor radiation and the correlation are shown in Table 1 (univariate analysis results for overall survival according to chemotherapy response and clinical variables).

<Example 2> Analysis of the effect of polymorphism on treatment outcome

Of the 96 selected polymorphisms, the CD155 rs1058402G>A (Ala67Thr, A67T) and CD226 rs763361C>T (Gly307Ser, G307S) polymorphisms were found to be associated with both chemotherapy response and OS. rs1058402G>A was found to be significantly associated with worse chemotherapy response and OS (adjusted OR [aOR] = 0.52, 95% CI = 0.27-0.99, P = 0.05 in the dominant model and adjusted for HR [aHR] = 1.55, 95% CI). = 1.12-2.14, P = 0.01; Table 2 and Figure 2A). Specifically, when the genotype of the rs1058402 polymorphism was GA or AA, the prognosis of chemotherapy response and OS was worse than that of GG. The effect of the rs1058402 polymorphism on PFS showed the same trend as that of OS, but was not statistically significant (aHR = 1.30, 95% CI = 0.96-1.76, P = 0.09 in the dominant model). In addition, the rs763361C>T polymorphism each showed significantly better chemotherapy response, OS and PFS compared to the rs1058402 polymorphism (adjusted aOR = 2.03, 95% CI = 1.10-3.75, P = 0.02; aHR = in the dominant model) 0.69, 95% CI = 0.51-0.94, P = 0.02; aHR = 0.73, 95% CI = 0.54-0.97, P = 0.03, Table 2 and Figure 2B). The genotyping results of the rs763361 polymorphism and the chemotherapy response, OS and PFS response to small cell lung cancer showed that when the genotype of the rs763361 polymorphism was CT or TT, the prognosis was better than that of CC (Table 2). ; Results of association analysis on chemotherapy response and survival outcomes according to genotype of CD155 rs1058402 and CD226 rs763361 polymorphisms).

We also analyzed survival outcomes in the extensive stage. In extended-stage SCLC, both rs1058402 and rs763361 were found to be associated with OS in univariate analysis (Log Rank P = 0.03 and 0.006 in the dominant model, respectively), and only the rs1058402 polymorphism was found to be associated with OS in multivariate analysis. (aHR = 1.75, 95% CI = 1.21-2.53, P = 0.003 in the dominant model). Also, PFS showed the same trend as OS, but was not statistically significant (in the dominant model, aHR = 1.34 for rs1058402, 95% CI = 0.94-1.90, P = 0.11, aHR = 0.79 for rs763361, 95% CI = 0.56- 1.10 , P = 0.16) (see Table 3).

The effect of polymorphic mutations in rs1058402 and rs763361 on chemotherapy response and OS did not differ according to clinical variables such as age and sex, except for the chemotherapy regimen for chemotherapy response (see Table 4). .

<Example 3> Analysis of effects on treatment results by the combination of rs1058402 and rs763361

The present inventors performed an analysis on the association of the combination of polymorphisms rs1058402 and rs763361 with the treatment of small cell lung cancer.

As a result, when the genotype of the rs1058402 polymorphism was GA or AA, and the genotype of the rs763361 polymorphism was CC, it was found that the chemotherapy response and OS were associated with poor outcomes. Thus, the polymorphic genotype was considered a bad genotype and showed a decrease in chemotherapy response as the number of bad genotypes increased (0 bad genotypes - 80.4% responders, 1 bad genotype - 71.0% responders, 2 bad genotypes) 61.3% responders; aOR = 0.52, 95% CI = 0.33-0.81, Ptrend = 0.004; Table 5). MST also appeared to decrease significantly with increasing number of bad genotypes (MST = 11.5 months for 0 bad genotypes, 9.5 months for 1 bad genotype, 7.2 months for 2 bad genotypes, aHR = 1.48, 95% CI = 1.19-1.84, Ptrend = 4 × 10-4 (Table 5 and Figure 2C). In addition, it was shown that PFS also decreased significantly with increasing number of bad genotypes (Ptrend = 0.009) (Table 5; analysis of association between chemotherapy response and survival analysis and the combination of rs1058402G > A and rs763361C > T genotypes).

Analysis of only patients with expanded stage showed that the combined effects of the rs1058402 and rs763361 polymorphisms were still significant on chemotherapy response, OS and PFS, respectively (aOR = 0.55, 95% CI = 0.33-0.93, Ptrend = 0.03; aHR = 1.55, 95% CI = 1.21-1.98, Ptrend = 6 × 10-4: aHR = 1.28, 95% CI = 1.07-1.59, Ptrend = 0.04).

<Example 4> Functional prediction through structural analysis of CD 155 rs1058402 G>A(A67T) polymorphism

In the rs1058402 G>A polymorphism of the CD155 gene, alanine to threonine at codon 67 of the CD155 protein is mutated. CD155 binds to CD226 of immune cells and acts as a co-stimulatory signal. A three-dimensional structural model was generated using PyMOL (https://pymol.org/2/) and analyzed whether these amino acid changes affect the function of CD155.

As a result, as shown in FIG. 3 , it was found that the distance from CD155 to S74 and G73 was shortened by the change of the 67th alanine of CD155 codon to threonine (from 6.3 Å to 2.9 Å and 4.4 Å to 3.3 Å, respectively). shortened to Å, Figures 3B, 3C). The short distance between T67 and S74 indicated that new hydrogen bonds were created (Fig. 3C). This change resulted in an increase in the distance between S74 of CD155 and N116 of CD226 (from 2.7 Å to 6.0 Å) and loss of hydrogen bonding (Figs. 3B, 3C). In addition, it was found that CD155 A67T affected other important interactions by distance extension (from 3.3 Å to 6.0 Å) between G70 of CD155 and E185 of CD226, resulting in loss of hydrogen bonds (Fig. 3D). Therefore, it was found that the increased distance and loss of hydrogen bonds could weaken the binding activity between CD155 and CD226. Therefore, the mutation of alanine to threonine at CD155 codon 67 interferes with the binding of CD155 and CD226 and reduces T cell activity, ultimately increasing the activity of cancer cells and decreasing the activity of inhibiting cancer cells, thereby further prolonging cancer progression in cancer patients. I knew it could make it worse.

So far, with respect to the present invention, the preferred embodiments have been looked at. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

<110> Kyungpook National University Industry-Academic Cooperation Foundation <120> Diagnostic methods for prognosis of small-cell lung cancer using CD155 and CD226 SNPs <130> PN2012-624 <160> 8 <170> KoPatentIn 3.0 <210> 1 <211> 500 <212> DNA <213> Artificial Sequence <220> <223> CD155 <400> 1 acaccccacg gtccaccggg gatccagggc cccagcgtgc aagtcgcgcc aggtgcccgg 60 gctcctggag cccctcccta tctagtccaa gaacgccccg ggtctgacac cttctcttcg 120 gttctccgca ggggacgtcg tcgtgcaggc gccccacccag gtgcccggct tcttgggcga 180 ctccgtgacg ctgccctgct acctacaggt gcccaacatg gaggtgacgc atgtgtcaca 240 gctgacttgg cgcggcatgg tgaatctggc agcatggccg tcttccacca aacgcagggc 300 cccagctatt cggagtccaa acggctggaa ttcgtggcag ccagactggg cgcggagctg 360 cggaatgcct cgctgaggat gttcgggttg cgcgtagagg atgaaggcaa ctacacctgc 420 ctgttcgtca cgttcccgca gggcagcagg agcgtggata tctggctccg agtgcttggt 480 gagcaggggg ttttggggag 500 <210> 2 <211> 1000 <212> DNA <213> Artificial Sequence <220> <223> CD226 <400> 2 catcttgcca acacaagagg aaagcctgca tgagagtgag gccaagcca gggaaatgga 60 tccaacactc tggaaaggga ttcagaagcc agtttatgcc catacttaat tctagacaca 120 acatttaagc cctggtaaat agcccttgcc caaagcttaa tctcccctgg atcattctgt 180 tatatgatac agtaagtttt cttttgtata taaaccagtt agagtcaggt tttctgaaac 240 agttctatga aaaatgattt taggtaatga agtatttttc ctatcaaagt aactcaattc 300 agacaactag tatctaaggt agaccttggg tagtggaaaa aaattgcata aagatccatg 360 catgagtaca taagagtcat tactaatgca ctcatgtcaa gaataagctt aaactctagt 420 ctttggtctg cgagagaagg ttggatagtt gacataaata tcctctcttg tatcatccat 480 ggattgattg gtaggttgac ggtagagatg ggacttctat agttattggg tgcctagaaa 540 gacaaacagc agagagtgtc aataattcac tgcatttcaa caactaatga agacattcaa 600 aacatacctc tcataaatgc aggcatgata tgggacaagt ttccattata attaatttga 660 taaaacattc tcattgtatt gaacttctgt tggaaatcat attcttgact gagatgcaac 720 agaaatctag tctagattaa tacttgctgt attgtgctat gaattacctg tttcaatgta 780 gaactccctc tcccaatgtt aagctatttt gataagaaca attactatca tttttatatg 840 gaaattctaa cacagtgtct gaactttaaa aaataagcaa cttaacatat aaaaagttaa 900 caaaactgac atcaagatga tattgtataa tcattaatct tctctaccgt aatcaagtca 960 ctttgaccac atatgtatgt atgtaacaga aaatatgcag 1000 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rs1058402 forward primer <400> 3 tacaggtgcc caacatggag 20 <210> 4 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> rs1058402 reverse primer <400> 4 gctgccagat tcaccat 17 <210> 5 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> rs763361 forward primer <400> 5 ccatggattg attggtag 18 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> rs763361 reverse primer <400> 6 gtttgtcttt ctaggcaccc 20 <210> 7 <211> 343 <212> PRT <213> Artificial Sequence <220> <223> CD155 <400> 7 Met Ala Arg Ala Met Ala Ala Ala Trp Pro Leu Leu Leu Val Ala Leu 1 5 10 15 Leu Val Leu Ser Trp Pro Pro Pro Gly Thr Gly Asp Val Val Val Gln 20 25 30 Ala Pro Thr Gln Val Pro Gly Phe Leu Gly Asp Ser Val Thr Leu Pro 35 40 45 Cys Tyr Leu Gln Val Pro Asn Met Glu Val Thr His Val Ser Gln Leu 50 55 60 Thr Trp Ala Arg His Gly Glu Ser Gly Ser Met Ala Val Phe His Gln 65 70 75 80 Thr Gln Gly Pro Ser Tyr Ser Glu Ser Lys Arg Leu Glu Phe Val Ala 85 90 95 Ala Arg Leu Gly Ala Glu Leu Arg Asn Ala Ser Leu Arg Met Phe Gly 100 105 110 Leu Arg Val Glu Asp Glu Gly Asn Tyr Thr Cys Leu Phe Val Thr Phe 115 120 125 Pro Gln Gly Ser Arg Ser Val Asp Ile Trp Leu Arg Val Leu Ala Lys 130 135 140 Pro Gln Asn Thr Ala Glu Val Gln Lys Val Gln Leu Thr Gly Glu Pro 145 150 155 160 Val Pro Met Ala Arg Cys Val Ser Thr Gly Gly Arg Pro Pro Ala Gln 165 170 175 Ile Thr Trp His Ser Asp Leu Gly Gly Met Pro Asn Thr Ser Gln Val 180 185 190 Pro Gly Phe Leu Ser Gly Thr Val Thr Val Thr Ser Leu Trp Ile Leu 195 200 205 Val Pro Ser Ser Gln Val Asp Gly Lys Asn Val Thr Cys Lys Val Glu 210 215 220 His Glu Ser Phe Glu Lys Pro Gln Leu Leu Thr Val Asn Leu Thr Val 225 230 235 240 Tyr Tyr Pro Pro Glu Val Ser Ile Ser Gly Tyr Asp Asn Asn Trp Tyr 245 250 255 Leu Gly Gln Asn Glu Ala Thr Leu Thr Cys Asp Ala Arg Ser Asn Pro 260 265 270 Glu Pro Thr Gly Tyr Asn Trp Ser Thr Thr Met Gly Pro Leu Pro Pro 275 280 285 Phe Ala Val Ala Gln Gly Ala Gln Leu Leu Ile Arg Pro Val Asp Lys 290 295 300 Pro Ile Asn Thr Thr Leu Ile Cys Asn Val Thr Asn Ala Leu Gly Ala 305 310 315 320 Arg Gln Ala Glu Leu Thr Val Gln Val Lys Glu Gly Pro Ser Glu 325 330 335 His Ser Gly Met Ser Arg Asn 340 <210> 8 <211> 336 <212> PRT <213> Artificial Sequence <220> <223> CD226 <400> 8 Met Asp Tyr Pro Thr Leu Leu Leu Ala Leu Leu His Val Tyr Arg Ala 1 5 10 15 Leu Cys Glu Glu Val Leu Trp His Thr Ser Val Pro Phe Ala Glu Asn 20 25 30 Met Ser Leu Glu Cys Val Tyr Pro Ser Met Gly Ile Leu Thr Gln Val 35 40 45 Glu Trp Phe Lys Ile Gly Thr Gln Gln Asp Ser Ile Ala Ile Phe Ser 50 55 60 Pro Thr His Gly Met Val Ile Arg Lys Pro Tyr Ala Glu Arg Val Tyr 65 70 75 80 Phe Leu Asn Ser Thr Met Ala Ser Asn Asn Met Thr Leu Phe Phe Arg 85 90 95 Asn Ala Ser Glu Asp Asp Val Gly Tyr Tyr Ser Cys Ser Leu Tyr Thr 100 105 110 Tyr Pro Gln Gly Thr Trp Gln Lys Val Ile Gln Val Val Gln Ser Asp 115 120 125 Ser Phe Glu Ala Ala Val Pro Ser Asn Ser His Ile Val Ser Glu Pro 130 135 140 Gly Lys Asn Val Thr Leu Thr Cys Gln Pro Gln Met Thr Trp Pro Val 145 150 155 160 Gln Ala Val Arg Trp Glu Lys Ile Gln Pro Arg Gln Ile Asp Leu Leu 165 170 175 Thr Tyr Cys Asn Leu Val His Gly Arg Asn Phe Thr Ser Lys Phe Pro 180 185 190 Arg Gln Ile Val Ser Asn Cys Ser His Gly Arg Trp Ser Val Ile Val 195 200 205 Ile Pro Asp Val Thr Val Ser Asp Ser Gly Leu Tyr Arg Cys Tyr Leu 210 215 220 Gln Ala Ser Ala Gly Glu Asn Glu Thr Phe Val Met Arg Leu Thr Val 225 230 235 240 Ala Glu Gly Lys Thr Asp Asn Gln Tyr Thr Leu Phe Val Ala Gly Gly 245 250 255 Thr Val Leu Leu Leu Leu Phe Val Ile Ser Ile Thr Thr Ile Ile Val 260 265 270 Ile Phe Leu Asn Arg Arg Arg Arg Arg Glu Arg Arg Asp Leu Phe Thr 275 280 285 Glu Ser Trp Asp Thr Gln Lys Ala Pro Asn Asn Tyr Arg Ser Pro Ile 290 295 300 Ser Thr Gly Gln Pro Thr Asn Gln Ser Met Asp Asp Thr Arg Glu Asp 305 310 315 320 Ile Tyr Val Asn Tyr Pro Thr Phe Ser Arg Arg Pro Lys Thr Arg Val 325 330 335

Claims (16)

A composition for a marker for the diagnosis and prediction of prognosis of small cell lung cancer, comprising at least one single nucleotide polymorphism selected from the group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226. According to claim 1,
The rs1058402G>A single nucleotide polymorphism of CD155 is located at the 250th base in the nucleotide sequence consisting of SEQ ID NO: 1,
The rs763361C>T single nucleotide polymorphism of CD226 is a marker composition for prognosis and prediction of small cell lung cancer, characterized in that it is located at the 500th base in the nucleotide sequence consisting of SEQ ID NO: 2. According to claim 1,
The small cell lung cancer is characterized in that the treatment using chemotherapy, marker composition for the diagnosis and prediction of the prognosis of small cell lung cancer. A composition for predicting survival prognosis of a patient with small cell lung cancer, comprising an agent capable of detecting one or more single nucleotide polymorphisms selected from the group consisting of rs1058402G>A single nucleotide polymorphism of CD155 and rs763361C>T single nucleotide polymorphism of CD226. 5. The method of claim 4,
The agent is a primer or probe capable of amplifying a polynucleotide consisting of 10 to 100 consecutive bases containing rs1058402G>A single nucleotide polymorphism of CD155 or rs763361C>T single nucleotide polymorphism of CD226 or a complementary polynucleotide thereof. A composition for predicting survival prognosis of small cell lung cancer patients, characterized in that. 5. The method of claim 4,
The rs1058402G>A single nucleotide polymorphism of CD155 is located at the 250th base in the nucleotide sequence consisting of SEQ ID NO: 1,
The composition for predicting survival prognosis of small cell lung cancer patients, characterized in that the rs763361C>T single nucleotide polymorphism of CD226 is located at the 500th base in the nucleotide sequence consisting of SEQ ID NO: 2. 5. The method of claim 4,
The agent comprises a primer pair consisting of the nucleotide sequences of SEQ ID NOs: 3 and 4 capable of detecting the rs1058402G>A single nucleotide polymorphism of CD155; Or a pair of primers consisting of the nucleotide sequences of SEQ ID NOs: 5 and 6 capable of detecting rs763361C>T single nucleotide polymorphism of CD226; 5. The method of claim 4,
The rs1058402G>A single nucleotide polymorphism of CD155 or the rs763361C>T single nucleotide polymorphism of CD226 is a chemotherapy response to an anticancer agent, overall survival (OS) or progression-free survival (PFS). A composition for predicting survival prognosis of small cell lung cancer patients, characterized in that it is related to. A kit for predicting survival prognosis of a patient with small cell lung cancer, comprising the composition according to claim 1 or 4. A microarray for predicting survival prognosis of a patient with small cell lung cancer, comprising the composition according to claim 1 or 4. A method of providing information for predicting survival prognosis of patients with small cell lung cancer, comprising the step of identifying the rs1058402G>A single nucleotide polymorphism of CD155 and the rs763361C>T single nucleotide polymorphism of CD226 with respect to the nucleic acid extracted from the sample. 12. The method of claim 11,
When the genotype of the rs1058402G>A polymorphism of CD155 is GG, it is characterized in that the survival prognosis is determined as a group having a higher survival prognosis than in the case of GA or AA. A method of providing information for predicting survival prognosis of patients with small cell lung cancer. 12. The method of claim 11,
When the genotype of the rs763361C>T polymorphism of CD226 is CT or TT, a method of providing information for predicting survival prognosis of small cell lung cancer patients, characterized in that the group has a higher survival prognosis than CC. 12. The method of claim 11,
GA or AA genotype of the rs1058402G>A polymorphism of CD155; Or, when the CC genotype of the rs763361C>T polymorphism of CD226 is 0, the chemotherapy response to the anticancer agent is 80.4%, and in the case of one, the chemotherapy response to the anticancer agent is 71.0%, and in the case of two, the chemotherapy response to the anticancer agent A method for providing information for predicting survival prognosis of patients with small cell lung cancer, characterized in that the response to therapy is 61.3%. 12. The method of claim 11,
In the rs1058402G>A polymorphism of CD155, when alanine (A), the 67th amino acid of CD155, is mutated to threonine (T), a chemotherapy response to an anticancer agent, overall survival (OS), or progression-free A method of providing information for predicting survival prognosis of small cell lung cancer patients, characterized in that the survival rate (Progression-free survival: PFS) is determined to be low. 16. The method of claim 15,
In the rs1058402G>A polymorphism of CD155, when alanine (A), the 67th amino acid of CD155, is mutated to threonine (T), the binding force between CD155 and CD226 is reduced, characterized in that T cell activity or NK cell activity is reduced, A method for providing information for predicting survival prognosis in patients with small cell lung cancer.

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