KR20200017195A - Method for Providing Information on Classification and Prognosis of Hematologic Malignancy - Google Patents

Abstract

The present invention relates to a method for providing information necessary for the classification and prognosis determination of hematologic malignancies, and to a composition used in the method. In the present invention, the level of an NGAL protein can be very useful in the classification and prognosis determination of various types of hematologic malignancies. In addition, by measuring the level of one or more proteins selected from the group consisting of IL-6, IL-1β, and TNF-α in addition to the NGAL protein, it is possible to provide very accurate and useful information for classification and prognosis determination of hematologic malignancies.

Description

Method for Providing Information on Classification and Prognosis of Hematologic Malignancy}

The present invention relates to a method for providing information necessary for the classification and prognosis of blood cancer, and to a composition used in the method.

The incidence of hematological tumor cancers is increasing as a cause of death diseases, and various causes include not only genetic factors of patients but also harmful environmental exposure and viral infection. Among these, infection and associated inflammation are recognized as one of risk factors for cancer.

If inflammation persists, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB), myeloid differentiation primary response gene 88, TLR4 (Toll-like receptor 4), and TNF (tumor necrosis factor) receptor-associated factors Variations accumulate in a variety of genes, including pre-malignant hematopoietic stem cell clones, which do not cause apoptosis or become hyperproliferative, leading to the development and progression of bone marrow malignancies. Is that.

Neutrophil gelatinase-associated lipocalin (NGAL) is a glycoprotein that has been characterized and isolated from granules of human neutrophils. The protein was isolated from human neutrophils as a 25-kDa glycoprotein covalently linked with matrix metalloproteinase (MMP) -9. NGAL is known to play a role as a biomarker of kidney damage, but recently it is involved in the regulation of iron-reactive genes for cell proliferation and is expressed in other tissues in response to various pathological conditions such as ischemia or tissue damage. It turns out to be. In addition, the expression of NGAL protein has been evaluated under various infectious and inflammatory diseases, and is involved in various processes such as apoptosis, apoptosis, inflammation, infection, and immune response. It is recognized as a family of fat-derived factors, adipokine.

CRP (C-reactive protein), interleukin (IL) -1β, IL-6 and tumor necrosis factor (TNF) -α are known as inflammatory factors. CRP is known as an acute protein of liver origin belonging to the petraxins superfamily. IL-1 and IL-6 are involved in the proliferation and differentiation of hematopoietic stem cells (HSCs) resulting in a decrease in the regenerative capacity of hematopoietic stem cells during chronic inflammation.

The patents and references mentioned herein are incorporated herein by reference to the same extent as if each document was individually and clearly specified by reference.

United States Patent Application Publication No. 2011/0081650 A1 Korean Patent Publication No. 10-2006-0095114

X. Leng, et al., Lipocalin 2 is required for BCR-ABL-induced tumorigenesis, Oncogene 27 (47) (2008) 6110-9.

The present inventors have tried to develop a method of providing useful information for the classification and prognosis of blood cancer by measuring the expression of biomarker proteins. As a result, the present invention was completed by experimentally confirming that by measuring the level of NGAL protein, it can provide not only the prognosis of blood cancer but also very useful information for its classification.

Accordingly, an object of the present invention is to provide a method for measuring the level of NGAL protein from a biological sample in order to provide information necessary for classification or prognosis of blood cancer.

Another object of the present invention is to provide a composition for classification or prognosis of blood cancer, including an agent capable of measuring the level of NGAL protein.

Other objects and technical features of the present invention are more specifically shown by the following detailed description, claims and drawings.

According to one aspect of the present invention, in order to provide information necessary for classification or prognosis of blood cancer, a method of measuring the level of NGAL (Neutrophil Gelatinase-Associated Lipocalin) protein from a biological sample is provided.

As used herein, the term "level of NGAL protein" refers to the amount in a biological sample of NGAL protein expressed from the gene encoding the protein.

As used herein, the term “blood cancer” refers to a cancer affecting the blood and lymphatic system and collectively refers to a cancer occurring in blood producing tissue (eg, bone marrow) or immune system cells.

According to one embodiment of the invention, the hematologic cancer is myeloproliferative neoplasm (MPN), acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and plasma cell neoplasm (plasma) cell neoplasms (PCN).

As used herein, the term "classifying" refers to the subclass of hematologic cancers that constitute hematologic cancer, more specifically myeloid growth tumor (MPN), acute myeloid leukemia (AML), and myelodysplasia. Syndrome (MDS) and plasma cell neoplasia (PCN) means that the cancer is classified into any one group.

As used herein, the term “myeloproliferative tumor (MPN) is a generic term for diseases in which the cells, such as red blood cells, white blood cells, platelets, and the like are excessively produced in the bone marrow (Tefferi A, Vainchenker W (February 2011). J. Clin. Oncol. 29 (5): 573-82).

According to one embodiment of the present invention, the myeloid proliferation tumor may include chronic myeloid leukemia (CML), erythrocytosis (polycythemia vera, PV), or thrombocythemia (ET).

As used herein, the term “acute myeloid leukemia (AML)” refers to cancerous mutations in precursor cells or stem cells in the early stages of differentiation of myeloid leukocytes, resulting in excessive division and It refers to a disease in which myeloblasts accumulate in bone marrow and accumulate in bone marrow.

As used herein, the term "myelodysplastic syndrome (MDS)" refers to a blood disease derived from abnormal hematopoietic stem cells characterized by proliferation of bone marrow, dysplasia of constituent cells, and inefficient hematopoiesis. As a disease that collectively refers to various types of diseases having these characteristics, it is known to occur in many elderly people. Myelodysplastic syndromes can be divided into primary myelodysplastic syndromes without a clear cause and secondary myelodysplastic syndromes after radiation therapy or chemotherapy for other diseases. In addition, it can be classified into several stages according to the ratio of blasts, immature cells of immature cells in the peripheral blood and bone marrow, and the degree of abnormality of the cells.

As used herein, the term "plasma cell neoplasia" (PCN) is a generic term for diseases caused by excessive proliferation of plasma cells. The plasma cell neoplasia (Multiple Myeloma, MM), its predecessor is a monoclonal gammopathy of undetermined significance (MGUS), SMM (smoldering multiple myeloma), single osteoblastic plasmacytoma (solitary) osseous plasmacytoma, solitary non-osseous plasmacytoma, or heavy chain disease.

As used herein, the term “determining the prognosis of blood cancer” is meant to include determining a particular prognosis of a blood cancer patient. Specific prognosis of the hematologic cancer includes, but is not limited to: (i) assessing the likelihood of metastasis to another organ, or risk of recurrence; (Ii) evaluating the stage of progression of blood cancer; (Iii) prognostic assessment of patients with hematological cancer if not treated; (Iii) evaluating the responsiveness of blood cancer patients to treatment methods (chemotherapy, radiation therapy; surgical operations); (Iii) diagnosis of the actual blood cancer patient's response to current or past treatment; (Iii) the selection of preferred methods for the treatment of hematological cancer patients; And (iii) prognostic determination of life expectancy of patients with hematological cancer (eg, prognostic determination of overall survival).

According to one embodiment of the invention, the methods of the invention may further measure the level of one or more proteins selected from the group consisting of IL-6, IL-1β, and TNF-α.

As used herein, the term "level of a protein of IL-6, IL-1β, or TNF-α" refers to a protein of IL-6, IL-1β, or TNF-α expressed from a gene encoding the protein. Means quantity.

According to one embodiment of the invention, (i) when the level of NGAL protein is lower than the control group can be classified as acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS).

According to another embodiment of the invention, (i) the level of NGAL protein is lower compared to the control, (ii) the level of IL-1β is lower than the control, and (iii) the level of TNF-α is compared with the control. In contrast, hematologic cancer may be classified as acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS).

According to another embodiment of the invention, (i) the level of NGAL protein is lower compared to the control, (ii) the levels of IL-6 and IL-1β are lower compared to the control, (iii) TNF-α Hematologic cancer may be classified as acute myeloid leukemia (AML) if the level of is higher than that of the control.

According to one embodiment of the present invention, (i) when the level of NGAL protein is higher than the control group hematological cancer can be classified as chronic myelogenous leukemia (CML) or erythrocytosis (PV).

According to another embodiment of the invention, (i) the level of NGAL protein is higher compared to the control, (ii) the level of IL-1β is lower than the control, and (iii) the level of TNF-α is compared with the control. In contrast, the hematologic cancer may be classified as chronic myelogenous leukemia (CML) or erythrocytosis (PV).

According to one embodiment of the invention, the biological sample is a supernatant of tissue, whole blood, serum, plasma, body fluids, urine, cells, cell debris or cell culture.

According to a more specific embodiment of the present invention, the biological sample is whole blood, serum, plasma, bone marrow-derived tissue, or bone marrow-derived cells, most specifically bone marrow-derived tissue or bone marrow-derived cells.

According to another aspect of the present invention, there is provided a composition for classification or prognosis of blood cancer, including an agent capable of measuring the level of NGAL protein.

According to one embodiment of the invention, the composition may further comprise an agent capable of measuring the level of one or more proteins selected from the group consisting of IL-6, IL-1β, and TNF-α.

According to one embodiment of the present invention, the hematologic cancer may be at least one selected from the group consisting of myeloproliferative tumor (MPN), acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and plasma cell neoplasia (PCN). .

According to one embodiment of the present invention, the myeloid proliferation tumor (MPN) may include chronic myeloid leukemia (CML), polycythemia vera (PV), or thrombocythemia (ET). .

According to one embodiment of the invention, the "agent that can measure the level of NGAL, IL-6, IL-1β, or TNF-α protein" may be an antibody that specifically binds to the protein.

For example, the agent includes, but is not limited to, an antibody or fragment thereof that can immunologically bind to the protein, a polypeptide derived from the protein, or a fragment thereof.

The antibody is a polyclonal or monoclonal antibody, preferably a monoclonal antibody. The antibody can be prepared by methods commonly practiced in the art, such as fusion methods (Kohler and Milstein, European Journal of Immunology, 6: 511-519 (1976)), recombinant DNA methods (US Pat. No. 4,816,56). Or phage antibody library methods (Clackson et al, Nature, 352: 624-628 (1991) and Marks et al, J. Mol. Biol., 222: 58, 1-597 (1991)). .

The method for measuring the level of a specific protein using the antibody can use a variety of immunological assays known in the art, for example, radioimmunoassay, radioimmunoprecipitation, immunoprecipitation, ELISA (enzyme-linked immunosorbent) assays, capture-ELISA, inhibition or competition assays, sandwich assays, flow cytometry, immunofluorescence staining and immunoaffinity purification.

Specifically, when the method of the present invention is carried out in an ELISA method, a specific embodiment of the present invention comprises the steps of (i) coating the sample to be analyzed on the surface of the solid substrate; (Ii) reacting the sample with an antibody against NGAL, IL-6, IL-1β, or TNF-α protein as the primary antibody; (Iii) reacting the resultant of step (ii) with the secondary antibody to which the enzyme is bound; And (iii) measuring the activity of the enzyme.

On the other hand, when the method of the present invention is carried out in a capture-ELISA mode, certain embodiments of the present invention are directed to (i) NGAL, IL-6, IL-1β, or TNF-α proteins as capturing antibodies. Coating the antibody on the surface of the solid substrate; (Ii) reacting the capture antibody with the sample; (Iii) reacting the result of step (ii) with a detecting antibody having a label that generates a signal and which specifically reacts with the modulator protein; And (iii) measuring the signal resulting from the label.

Measurement of the final enzyme activity or signal in the ELISA method and the capture-ELISA method can be carried out according to various methods known in the art.

The features and advantages of the present invention are summarized as follows:

(i) The present invention relates to a method for providing information necessary for classification and prognosis of blood cancer, and a composition used in the method.

(Ii) In the present invention, the level of NGAL protein can be very useful for the classification and prognosis of various types of blood cancer.

(Iii) The present invention provides highly accurate and useful information for the classification and prognosis of blood cancer by further measuring the level of one or more proteins selected from the group consisting of IL-6, IL-1β, and TNF-α in addition to NGAL proteins. Can provide.

1 shows the results of a combination assay of NGAL and inflammatory cytokines IL-1β, IL-6, and TNF-α via Western blot.

Example

Experiment method

1. Preparation of samples

Bone marrow (BM) aspirates and peripheral blood (PB) samples were obtained from 41 patients with hematologic malignancy at Korea University Ansan Hospital. Patients who collected the samples were treated with myeloid growth tumor (MPN, n = 12), acute myeloid leukemia (AML, n = 5), myelodysplastic syndrome (MDS, n = 13) and plasmacytocin according to WHO diagnostic criteria. Groups of patients classified for each disease of living organisms (plasma cell neoplasms, PCN, n = 6). As a control, samples of lymphoma patients (n = 5) without bone marrow (BM) involvement were used. Patients with symptoms of infection, inflammatory disease or kidney disease were excluded from the experimental and control groups. After centrifugation (3000 rpm, 10 minutes), the sample was placed in a tube containing ethylene-diamine-tetra acetic acid (EDTA), from which a supernatant of PB plasma and BM aspirate was obtained and analyzed. Store at 80 ° C. PB and BM aspirate samples were analyzed simultaneously. Plasma levels of NGAL, IL-1b, IL-6, and TNF-α in PB and BM aspirate samples were measured using an enzyme-linked immunosorbent assay (ELISA) and western blot on an automated platform.

2. Design of Experiment

Supernatant samples of a total of 41 PB plasma and BM aspirates were analyzed by CRP and NGAL tests on an automated platform. Of these samples, 20 samples were analyzed for NGAL, IL-6, IL-1β and TNF-α by Western blot. NGAL analysis used monomeric specific monoclonal antibodies for western blot and immunoassay of automated platforms. Analysis results were assessed for association with clinical features including bone marrow features.

3. Immunoassay of NGAL and CRP

Particle-enhanced turbidity immunoassay using a Cobas 8000 (Roche Diagnostics, Rotkreuz, Switzerland) automated instrument for PB plasma and BM supernatants using mouse monoclonal anti-NGAL (human) antibodies (BioProto Diagnostics, Gentofte, Denmark) Was performed. Specifically, 3 μl of sample and 150 μl of reaction buffer were reacted for 5 minutes and further reacted for 5 minutes using 50 μl suspension of immunoparticles to determine the level of NGAL. The concentration of NGAL was calculated from absorbance changes based on a calibration curve prepared using calibration results of known concentrations of NGAL from 50 ng / mL to 3000 ng / mL. CRP measurements were made using an immunoturbidimetric assay assay using Cobas 8000 (Roche Diagnostics GmbH, Penzberg, Germany).

4. Western Blot

After taking a portion of the BM supernatant and PB plasma, the total protein amount was measured using the Quick Start Bradford Protein Assay Kit (Bio-Rad, Inc., Hercules, Calif., USA). About 30 μg of total protein was loaded into each lane on a 12% SDS-polyacrylamide gel. Proteins are separated by electrophoresis for 4 hours to 6 hours at 50 V in tris-glycine buffer, and PVDF membrane (Millipore Corporation) for 6 hours at 100 mA at 4 ° C. in transfer buffer. , MA, USA). Each blot was blocked for 90 minutes with stirring at room temperature in 5% skim milk in tris-buffered saline (TBS) and 0.1 Tween 20 (TBST). Primary antibodies were diluted in TBST with 3% skim milk: anti-NGAL antibody (1: 1000, abcam, Cambridge, UK), anti-IL-1β antibody (1: 500, Santa Cruz, TX, USA) , Anti-IL-6 antibody (1: 500 dilution, Santa Cruz), anti-TNF-α antibody (1: 1000, Cell Signaling Technology, MA, USA) and anti-Transferrin antibody (1: 1000, Santa Cruz, TX , USA). The blot was reacted overnight at 4 ° C. with each primary antibody, washed with TBST, and then horseradish peroxidase-conjugated secondary antibody (1: 2000, Santa Cruz, TX, USA) Reaction at room temperature for 90 minutes. After thorough washing with TBST, blots were immersed in NBT / BCIP substrate (Roche Applied Science) until the results were visualized. Immunocomplexes were detected using the ECL system. The relative intensity of each blot was analyzed using Image J analysis software (National Institutes of Health, USA).

5. Statistical Method

NGAL and CRP levels of PB and BM samples were analyzed using Paired t-test or Mann-Whitney U test. Mann-Whitney U test was used to compare successive variations between disease groups and controls. Statistical significance was defined as P <0.05 or P <0.01 or P <0.001. All statistical analyzes were performed using SPSS version 21.0 (SPSS, Chicago, USA).

Experiment result

1. Patient Characteristics Analysis

BM and PB samples were obtained in pairs from a total of 41 patients with an average age of 63 years (35-88 years) and males 53.7% (22/41) and females 46.3% (19/41). Patients who obtained the samples were myeloid growth tumors (MPN, n = 12), acute myeloid leukemia (AML, n = 5), myelodysplastic syndromes (MDS, n = 13), plasma cell neoplasia (PCN, n = 6 ), And patients diagnosed with lymphoma without bone marrow involvement (n = 5, control). MPN samples (n = 12) consisted of chronic myelogenous leukemia (CML, n = 4), erythrocytosis (polycythemia vera, PV, n = 7), and thrombocythemia (ET, n = 1).

2. Relationship between NGAL and CRP levels in hematologic malignancies in peripheral blood (PB) and bone marrow (BM)

Analysis of NGAL levels in PB and BM samples (n = 41) revealed 163.0 ± 258.3 and 413.1 ± 616.2 ng / mL (mean ± SD), respectively. The NGAL level (PB-NGAL) in the PB sample showed the highest value (329.0 ± 417.1) (mean ± SD) in the MPN group and the lowest value (30.8 ± 21.2) (mean ± SD) in the AML group. . Similarly, the NGAL level (BM-NGAL) in the BM sample also showed the highest value (1184.9 ± 1045.0) (mean ± SD) in the MPN group and the lowest value (44.8 ± 52.1) (mean ± SD) in the AML group. ) (See Table 1). These results are statistically significant.

In addition, CRP expression levels in PB samples and BM samples (n = 41) were 1.195 ± 2.160 and 1.187 ± 2.087 mg / dL (mean ± SD), respectively (see Table 1). The CRP expression level (PB-CRP) in the PB sample showed the highest value (3.547 ± 2.472) (mean ± SD) in the AML group and the lowest value (0.446 ± 0.844) (mean ± SD) in the PCN group. Indicated. Similarly, CRP expression levels (BM-CRP) in BM samples showed the highest value (3.672 ± 2.782) (mean ± SD) in the AML group and the lowest value (0.422 ± 0.796) in the PCN group. (See Table 1). For the total sample and each disease group, the expression level of CRP showed no statistically significant difference in PB and BM.

From the above results, the NGAL showed the highest level in the MPN group, but the lowest in the AML group. This trend was more pronounced in BM samples than in PB samples. In contrast, the CRP showed the highest level in the AML group, but the low value in the MPN group, in contrast to the NGAL results. That is, among the five disease groups, they showed opposite patterns on NGAL and CRP levels. This suggests that the two substances will play opposite roles in the development of the inflammatory response in blood cancers.

NGAL expression levels and CRP expression levels in PB and BM samples showed different patterns according to each disease group (MPN, AML, MDS, PCN) of hematologic tumors.

In the MPN group, the level of NGAL was determined to be higher than the control, and this NGAL expression level pattern was more pronounced in the BM sample (BM-NGAL) than in the PB sample (PB-NGAL) (P = 0.001).

For CML in the MPN group, BM-NGAL levels (2458.1 ± 659.6 ng / mL) were higher than PB-NGAL levels (595.0 ± 692.3 ng / mL). In the PV of the MPN group, the BM-NGAL level (578.4 ± 393.7 ng / mL) was higher than the PB-NGAL level (192.7 ± 91.5 ng / mL). However, in the AML, MDS, and PCN groups, there was no statistically significant difference in NGAL levels in BM and PB. The results of measurement of CRP levels in PB and BM samples (n = 41) were 1.195 ± 2.160 and 1.187 ± 2.087 mg / dL, respectively, and there was no statistically significant difference between PB and BM samples. There was no statistically significant difference between BM samples (Table 1).

group NGAL (ng / ml) (mean ± SD) p-value CRP (mg / ml) (mean ± SD) p-value PB BM PB BM MPN (n = 12) 329.0 ± 417.1 1184.9 ± 1045.0 0.001 * 0.449 ± 1.145 0.430 ± 1.095 0.665 AML (n = 5) 30.8 ± 21.2 44.8 ± 52.1 0.834 3.547 ± 2.472 3.672 ± 2.782 0.602 MDS (n = 13) 69.6 ± 48.7 106.0 ± 90.4 0.158 1.551 ± 2.886 1.463 ± 2.563 0.939 PCN (n = 6) 179.6 ± 133.4 392.2 ± 307.8 0.078 0.446 ± 0.844 0.422 ± 0.796 0.631 Control
(n = 5) 85.0 ± 22.3 203.1 ± 78.9 0.016 * 0.609 ± 0.886 0.720 ± 1.088 1.000 Total
(n = 41) 163.0 ± 258.3 413.1 ± 616.2 0.002 * 1.195 ± 2.160 1.187 ± 2.087 0.865

*: Statistically significant (p <0.05, by paired t-test or Mann-Whitney U test).

3. Comparison of BM-NGAL levels in each disease group through automated NGAL testing

Compared to the control BM-NGAL level (203.1 ± 78.9 ng / mL), the BM-NGAL level of MPN was significantly higher (1184.9 ± 1045.0 ng / mL; P <0.01), but the BM-NGAL level of AML (44.8 ± 52.1 ng / mL; P <0.05) and BM-NGAL (106.0 ± 90.4 ng / mL; P <0.05) of MDS were significantly lower. For the MPN group, the CML (2458.1 ± 659.6 ng / mL) and PV (578.4 ± 393.7 ng / mL) groups had significantly higher BM-NGAL levels compared to the control (Table 2).

4. Comparison of PB-NGAL level of each disease group through automated NGAL test

Compared to the control PB-NGAL level (85.0 ± 22.3 ng / mL), the PB-NGAL level (329.0 ± 417.1 ng / mL; P <0.01) of the MPN group was significantly higher, but the PB-NGAL level of the AML group ( 30.8 ± 21.2 ng / mL; P <0.05) was significantly lower. However, unlike in BM samples, PB-NGAL levels did not show statistically significant differences in MDS groups and controls. In the MPN group, the PB-NGAL level (595.0 ± 692.3 ng / mL) of the CML group and the PB-NGAL level (192.7 ± 91.5 ng / mL) of the PV group were significantly higher compared to the control (Table 2).

group NGAL (ng / ml)
(Mean ± SD) p-value group NGAL (ng / ml)
(Mean ± SD) p-value PB MPN (n = 12) 329.0 ± 417.1 0.006 ^* BM MPN (n = 12) 1184.9 ± 1045.0 0.004 ^* AML (n = 5) 30.8 ± 21.2 0.016 ^* AML (n = 5) 44.8 ± 52.1 0.028 ^* MDS (n = 13) 69.6 ± 48.7 0.218 MDS (n = 13)  106.0 ± 90.4 0.034 ^* PCN (n = 6) 179.6 ± 133.4 0.100 PCN (n = 6)  392.2 ± 307.8 0.201 Control (n = 5) 85.0 ± 22.3 Control (n = 5) 203.1 ± 78.9

*: Statistically significant (p <0.05, by paired t-test or Mann-Whitney U test).

5. Comparison of CRP level of each disease group through automated immunoassay

Compared to the control BM-CRP level (0.720 ± 1.088 mg / dL), the BM-CRP level (3.672 ± 2.782 mg / dL; P <0.05) in the AML group was significantly higher. The other disease groups did not show significant differences compared to the control. PB samples did not show any significant differences from the control group in all disease groups.

6. Combination assay of NGAL and inflammatory cytokines IL-1β, IL-6, and TNF-α via Western blot

Western blotting methods measured NGAL, IL-6, IL-1β and TNF-α levels in BM and PB samples and then compared their relative levels with each other.

In the CML group (P <0.01) and PV group (P <0.05), BM-NGAL levels were significantly higher than PB-NGAL levels. In the AML group, BM-IL-6 levels were higher than PB-IL-6 levels (P <0.05). For IL-1β, there was no statistically significant difference between BM-IL-1β and PB-IL-1β levels in all disease groups. The BM-TNF-α levels in the CML group (P <0.05) and the MDS group (P <0.01) were significantly higher compared to the PB-TNF-α levels.

In the CML group (P <0.001) and PV group (P <0.05), BM-NGAL levels increased significantly compared to the control. In contrast, BM-NGAL levels were significantly decreased in the MDS group (P <0.01). In PB-NGAL, only the PV group showed a significantly higher level (P <0.05) compared to the control (A in FIG. 1).

For BM-IL-6, no disease group showed significant difference from the control group. In the case of PB-IL-6, the AML group showed significantly lower levels (P <0.05) compared to the control group (B in FIG. 1).

For BM- and PB-IL-1β, all of the disease groups showed significantly lower levels compared to the control group (P <0.001) (FIG. 1C).

For BM-TNF-α, all disease groups showed higher levels compared to the control group (P <0.01) (Panel D of FIG. 1). PB-TNF-α showed significantly higher levels in the AML group (P <0.001), PV and MDS groups (P <0.01) compared to the control (D in FIG. 1).

Specific embodiments described herein are meant to represent preferred embodiments or examples of the present invention, whereby the scope of the present invention is not limited. It will be apparent to those skilled in the art that variations and other uses of the invention do not depart from the scope of the invention described in the claims.

Claims (10)

A method of measuring the level of Neutrophil Gelatinase-Associated Lipocalin (NGAL) protein from a biological sample to provide information necessary for classification or prognosis of blood cancer.
The method of claim 1, further comprising measuring the level of one or more proteins selected from the group consisting of IL-6, IL-1β, and TNF-α.
According to claim 1 or 2, wherein the hematologic cancer is acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), plasma cell neoplasms (PCN), chronic myelogenous leukemia (chronic myeloid leukemia, CML), polycythemia vera (PV), and thrombocythemia (ET).
4. The method of claim 3, wherein (i) the level of NGAL protein is lower compared to the control, (ii) the level of IL-1β is lower than the control, and (iii) the level of TNF-α is compared with the control. The blood cancer is classified as acute myelogenous leukemia (AML) or myelodysplastic syndrome (MDS).
4. The method of claim 3, wherein (i) the level of NGAL protein is higher compared to the control, (ii) the level of IL-1β is lower than the control, and (iii) the level of TNF-α is compared with the control. And in contrast, classifying the hematologic cancer as chronic myelogenous leukemia (CML) or erythrocytosis (PV).
The method of claim 1 or 2, wherein the biological sample is whole blood, serum, plasma, bone marrow-derived tissue, or bone marrow-derived cells.
A composition for classification or prognosis of blood cancer, comprising an agent capable of measuring the level of NGAL protein.
8. The composition of claim 7, further comprising an agent capable of measuring one or more levels selected from the group consisting of IL-6, IL-1β, and TNF-α.
The composition of claim 7 or 8, wherein the agent capable of measuring the level of NGAL, IL-6, IL-1β, or TNF-α protein is an antibody that specifically binds to the protein.
The method of claim 7 or 8, wherein the hematologic cancer is acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), plasma cell neoplasia (PCN), chronic myelogenous leukemia (CML), erythrocytosis (PV), and At least one composition selected from the group consisting of thrombocytopenia (ET).

Images