JP2023049325A - Method of predicting prognosis of lung adenocarcinoma patient based on expression of basic fibroblast growth factor in stroma - Google Patents

Abstract

To provide a method of predicting prognosis of a lung adenocarcinoma patient and a method of treating the lung adenocarcinoma patient.SOLUTION: A method of predicting prognosis of a lung adenocarcinoma patient is provided, comprising detecting the expression of the basic fibroblast growth factor (bFGF) in a lung adenocarcinoma tissue obtained from the lung adenocarcinoma patient, where positive bFGF indicates good prognosis or possibility thereof while negative bFGF indicates bad prognosis or possibility thereof. Also provided are pharmaceutical compositions for use in treating lung adenocarcinoma patients, including therapeutically effective amounts of anti-vascular endothelial growth factor (VEGF) drugs.SELECTED DRAWING: None

Description

The present disclosure relates to methods of predicting the prognosis of lung adenocarcinoma patients based on the expression of basic fibroblast growth factor in and methods of treating lung adenocarcinoma in subjects determined to have a poor prognosis.

In 2018, the number of cancer deaths in Japan was 1.36 million, of which 370,000 (27%) died of cancer. In 2018, 980,000 people were newly diagnosed with cancer, and accurately diagnosing cancer is a social issue.

Non-Patent Document 1 examines the relationship between stage II and III non-small cell lung cancer and FGF2 expression and discloses that FGF2 negativity correlates with improved overall survival (OS). Non-Patent Document 2 examines the relationship between lung squamous cell carcinoma and FGF2 expression, and discloses that there is no clear correlation between FGF2 expression and OS. Non-Patent Document 3 show that overexpression of bFGF is associated with poor prognosis in resectable non-small cell lung cancer.

Int. J. Radiation Oncology Biol. Phys. , 82(1): 442-447, 2012 Int J Clin Exp Pathol. , 8(9): 9760-9771, 2015 PLoS One, 11(1): e0147374. 2016

The present disclosure provides methods of predicting the prognosis of lung adenocarcinoma patients based on the expression of basic fibroblast growth factor (bFGF) in and methods of treating lung adenocarcinoma in subjects determined to have a poor prognosis. I will provide a.

The present inventors found that the expression of bFGF in stromal cells in lung adenocarcinoma biopsies obtained from lung adenocarcinoma patients affects the prognosis of the patients after cancer resection. .

According to the present invention, the following inventions are provided.
[1] A method for predicting the prognosis of a patient with lung adenocarcinoma, comprising:
detecting expression of basic fibroblast growth factor (bFGF) in lung adenocarcinoma tissue obtained from a patient with lung adenocarcinoma, wherein positive bFGF indicates a favorable prognosis or A method in which the possibility is indicated and, when negative, the prognosis is or is indicated to be poor.
[2] The method according to [1] above, wherein the cancer patient is a patient after cancer resection.
[3] The method of [1] or [2] above, wherein the level of expression is determined based on the ratio of positive cells to cancer cells.
[4] The method according to any one of [1] to [3] above, wherein the expression is protein expression.
[5] The method according to any one of [1] to [4] above, wherein the prognosis is at least one selected from the group consisting of recurrence rate and survival rate.
[6] The method according to any one of [1] to [5] above, wherein the lung adenocarcinoma is an early stage cancer selected from the group consisting of stages I and II.
[7] A pharmaceutical composition for use in treating a patient with lung adenocarcinoma, comprising:
comprising a therapeutically effective amount of an anti-vascular endothelial growth factor (VEGF) agent;
A pharmaceutical composition, wherein the lung adenocarcinoma patient is a patient evaluated to have a poor prognosis by the method according to any one of [1] to [6] above.
[8] The pharmaceutical composition of [7] above, wherein the anti-VEGF drug is bevacizumab.
[9] A method of treating cancer in a subject with cancer, comprising:
Predicting the prognosis of the subject by the method according to any one of [1] to [6] above;
A pharmaceutical composition comprising an anti-angiogenic agent for use in a method comprising administering a therapeutically effective amount of the anti-angiogenic agent to a subject predicted to have a poor prognosis.
[10] The method according to any one of the above, wherein a positive evaluation is made when the bFGF expression level is equal to or higher than a predetermined value, and a negative evaluation is made when it is less than the predetermined value.
[11] Any of the above-described methods, wherein the expression level of bFGF is evaluated as positive when it exceeds a predetermined value, and is evaluated as negative when it is equal to or less than the predetermined value.

FIG. 1 is an immunohistochemical staining image for bFGF of a cancer tissue biopsy obtained from a lung adenocarcinoma patient. FIG. 2 shows the recurrence-free survival of the stage I patient group and the stage II-IV patient group. FIG. 3 shows the recurrence-free survival of stage I and II patient groups and stage III and IV patient groups. FIG. 4 shows the survival times of the stage I patient group and the stage II-IV patient group. FIG. 5 shows the survival times of stage I and II patient groups and stage III and IV patient groups. FIG. 6 shows recurrence-free survival times of bFGF-positive and -negative cases in all cases. FIG. 7 shows the recurrence-free survival of bFGF-positive and -negative cases in the stage I and II patient groups. FIG. 8 shows recurrence-free survival times of bFGF-positive and -negative cases in the stage I patient group. FIG. 9 shows survival times of bFGF-positive and -negative cases in all cases. FIG. 10 shows survival times of bFGF-positive and -negative cases in stage I and II patient groups. FIG. 11 shows survival times of bFGF-positive and -negative cases in the stage I patient group.

Detailed description of the invention

<Definition>
As used herein, a "subject" can be a mammal, preferably a human. The subject can be one suffering from or at risk for a tumor or cancer. Subjects can include subjects 70 years of age or older and less than 70 years of age. Subjects may include smokers (eg, BI index greater than or equal to 700) and non-smokers.

As used herein, "cancer" is a disease characterized by uncontrolled growth of cells. Cancer cells can spread locally or spread throughout the body through the bloodstream or lymphatic system. Non-limiting examples of cancer include, for example, solid tumors. Non-limiting examples of cancer include, for example, hematopoietic malignancies. Solid tumors also include, for example, lung cancer. As used herein, "cancer tissue" is tissue that contains cancer. Cancer tissue can be obtained by biopsy and cancer resection. As used herein, "prognosis" can mean recurrence rate (or time to recurrence) and survival rate of cancer. Recurrence includes recurrence at the primary focus and recurrence after metastasis. The recurrence rate can be the recurrence rate after a period of treatment. The period of time is, for example, 1 year, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, or 15 years. It can be a period of years, or any of these periods or longer.

As used herein, "lung adenocarcinoma" is a type of lung cancer. Lung cancer is roughly classified into small cell lung cancer and non-small cell lung cancer, and non-small cell lung cancer is roughly classified into three cancers including lung adenocarcinoma. Non-small cell lung cancer accounts for about 80% of lung cancer, and lung adenocarcinoma accounts for about 60% of non-small cell lung cancer.

As used herein, "gene expression" or similar expressions refers to both messenger RNA (mRNA) and protein expression from genes present on the genome of a cell. mRNA can be obtained by extracting total RNA from cells by methods well known to those skilled in the art, for example, by synthesizing the first strand of cDNA using the poly A sequence unique to mRNA, followed by amplification (for example, polymerase chain reaction). ) can be detected. Proteins can be detected using antibodies (eg, polyclonal or monoclonal antibodies) that specifically bind to the protein.

As used herein, "positive" in the context of gene expression in cells means detection of mRNA or protein at detectable levels, unless otherwise specified. Also, "negative" means that no detectable levels of mRNA or protein are detected, unless otherwise specified. Herein, in order to further clarify the classification of "positive" and "negative" in the context of gene expression in tissues, a threshold is set, and those showing an expression level above the threshold are defined as "positive", Those showing an expression level below the threshold are indicated as "negative". Thus, in the present specification, the determination criteria for "positive" gene expression at the cellular level and "positive" gene expression at the tissue level are different.

As used herein, expression level refers to the amount of mRNA or protein expressed in a tissue or given cell population. Expression levels of mRNA can be quantified by various methods well known to those skilled in the art for quantifying mRNA, such as quantitative PCR. Protein can be quantified by methods well known to those skilled in the art, such as immunohistochemical staining (IHC). In IHC, antibodies bind to proteins in tissue sections and can be colored with a dye. The intensity of color development corresponds to the amount of protein expression. Therefore, the amount of protein expression can be quantified by the intensity of color development. Also, in IHC, the expression level of a protein can be quantified based on the number of expressing cells or the percentage of cells instead of the intensity of color development. Whether the protein expression level is quantified based on the intensity of color development or based on the number of expressing cells or the cell ratio is selected according to the situation and purpose.

As used herein, "basic fibroblast growth factor" (bFGF) is a growth factor belonging to the FGF family. bFGF is also called FGF2. In addition to angiogenesis, bFGF has been suggested to be involved in the development of nervous system, lung, muscle, bone, skin, and the like. Human bFGF can have, for example, the amino acid sequence registered under NCBI Reference Sequence: NP_001997.5. Human bFGF may have 80% or more, 85% or more, 90% or more, or 95% or more amino acid sequence identity with the amino acid sequence registered under NCBI Reference Sequence: NP_001997.5 .

As used herein, "antibody" means an immunoglobulin. Antibodies can be of various isotypes, eg, IgG. Antibodies may preferably be monoclonal antibodies. Antibodies can be human chimeric antibodies, humanized antibodies, or human antibodies. Human chimeric antibodies can be made by replacing the constant regions of non-human antibodies with those of human antibodies. A humanized antibody can be made by replacing the six CDRs of a human antibody with the corresponding six CDRs of a non-human antibody. Human antibodies can be produced using animals (eg, mice) in which at least the heavy chain variable region of an immunoglobulin has been replaced with the corresponding region from the human locus. When the constant region is non-human, a human antibody can be obtained by replacing the constant region with the amino acid sequence of a human antibody. As used herein, the antibody may preferably be a humanized antibody. As used herein, the antibody may preferably be a human antibody. An antibody has a signal peptide when it is produced intracellularly, but the signal peptide is excised when it is secreted extracellularly. Therefore, the antibody does not require a signal peptide when administered as a pharmaceutical.

As used herein, "CDRs" are the complementarity determining regions present in the heavy and light chain variable regions of an antibody. There are three each in the heavy and light chain variable regions, designated from the N-terminus as CDR1, CDR2 and CDR3. CDRs are, for example, numbered according to the numbering of Kabat et al. can be determined based on

As used herein, "compete" means competing with other binding antibodies for binding to an antigen. Competition can occur when the binding sites of two antibodies overlap for an antigen. Such antibodies can be obtained by immunization with the epitope as described above and/or by competition assays to determine whether the binding of one antibody to the antigen is reduced by the other antibody. Obtainable. Competing antibodies include antibodies that compete with each other.

As used herein, "antibody-drug conjugate" (hereinafter also referred to as "ADC") means a substance in which an antibody and a cytotoxic agent are linked. In ADCs, the antibody and cytotoxic agent can be linked via a suitable linker. Chemotherapeutic agents, radioisotopes, and toxins can be used as cytotoxic agents. ADCs also include conjugates of antigen-binding fragments of antibodies and drugs.

As used herein, an "antigen-binding fragment of an antibody" refers to a fragment of an antibody that maintains antigen-binding ability. Antigen-binding fragments include, for example, Fab, Fab', F(ab') ₂ , Fv, scFv (single-chain Fv), diabodies, sc(Fv) ₂ (single-chain (Fv) ₂ ). For example, papain digestion of antibodies can yield Fabs. Alternatively, the antibody can be digested with pepsin to give F(ab') ₂ , which can be further reduced to give Fab'. Other antigen-binding fragments of antibodies can also be produced by methods well known to those skilled in the art. Antigen-binding fragments of such antibodies can be used in the present invention.

<Inspection method and prediction method of the present disclosure>
According to this disclosure:
(A) A method for predicting the prognosis of a cancer patient, comprising:
A step of detecting the expression of FGF (particularly bFGF) in cancer tissue obtained from a cancer patient,
A method is provided.

In the above method (A), positive bFGF expression in cancer tissue can predict good prognosis or possibility of good prognosis. Alternatively, in the above method, negative bFGF expression in cancer tissue can be predicted to have a poor prognosis or a possibility thereof. In the above method, positive bFGF expression in cancer tissue can predict that the prognosis is good or likely to be good, and negative expression of bFGF in cancer tissue can predict the prognosis. can be predicted to be bad or likely to be bad.

In one embodiment, bFGF positive or negative can be determined based on bFGF protein or mRNA levels in tissue (or sections or cell lysates thereof).

In one aspect, bFGF levels can be determined using an antibody that binds to bFGF. The tissue (or its section or cell lysate) is brought into contact with an antibody that binds to bFGF under conditions where a complex between the bFGF protein and the antibody is formed, and when the complex is formed, the bFGF protein is detected. Detection can be performed by methods well known to those skilled in the art, such as ELISA, Western blotting and protein array methods. Based on this detected amount, it can be determined that the cancer tissue is bFGF positive or negative. In a preferred embodiment, bFGF-positive or negative cancer tissue can be confirmed by immunohistochemical staining. Protein levels of bFGF can be measured using techniques well known to those of skill in the art.

In one aspect, bFGF mRNA levels can be measured using techniques well known to those of skill in the art. For example, bFGF mRNA levels can be determined by quantitative assays such as quantitative PCR, digital PCR, microarrays, or next-generation sequencing.

According to the present disclosure, in the above method (A), the bFGF protein expression level is determined to be bFGF positive when it is equal to or greater than the first predetermined value, and when it is less than the first predetermined value can be determined to be bFGF-negative. Further, in the above method (A), if the expression level of bFGF mRNA is equal to or greater than a second predetermined value, it is determined to be bFGF positive, and if it is less than the second predetermined value, bFGF can be determined to be negative.

The first predetermined value can be the maximum bFGF protein expression level or less, the third quartile value or less, the mean value or less, or the median value or less in cancer patients with good prognosis. The predetermined value can be the minimum bFGF protein expression level or more, the first quartile or more, the mean value or more, or the median value or more of the bFGF protein expression level in the cancer patient group with poor prognosis. The first predetermined value is the maximum value or less, the third quartile value or less, the mean value or less, or the median value or less of the bFGF protein expression level in the cancer patient group with good prognosis, and the prognosis is poor. The bFGF protein expression level in the cancer patient group can be at least the minimum value, at least the first quartile value, at least the average value, or at least the median value.

The second predetermined value can be the maximum bFGF protein expression level or less, the third quartile value or less, the mean value or less, or the median value or less in cancer patients with good prognosis. The predetermined value can be the minimum bFGF mRNA expression level or more, the first quartile or more, the mean value or more, or the median value or more of the bFGF mRNA expression level in the cancer patient group with poor prognosis. The first predetermined value is the maximum value or less, the third quartile value or less, the mean value or less, or the median value or less of the bFGF mRNA expression level in the cancer patient group with good prognosis, and the prognostic The expression level of bFGF protein mRNA in the group of patients with unfavorable cancer can be at least the minimum value, at least the first quartile value, at least the average value, or at least the median value.

In the above, whether the prognosis is good or bad can be determined by a doctor. Good or bad prognosis can be determined, for example, by length of recurrence-free survival or length of survival. For example, recurrence-free survival after cancer resection is 1,000 hours or longer, 1,100 hours or longer, 1,200 hours or longer, 1,300 hours or longer, 1,400 hours or longer, 1,500 hours or longer, 1,600 hours or more, 1,700 hours or more, 1,800 hours or more, 1,900 hours or more, 2,000 hours or more, 2,100 hours or more, 2,200 hours or more, 2,300 hours or more, 2, Patients with a good prognosis were determined to be ≥400 hours, ≥2,500 hours, ≥2,600 hours, ≥2,700 hours, ≥2,800 hours, ≥2,900 hours, or ≥3,000 hours. Patients below that (those who were not determined to have a good prognosis) can be determined to have a poor prognosis. Or, for example, survival time after cancer resection is 1,000 hours or longer, 1,100 hours or longer, 1,200 hours or longer, 1,300 hours or longer, 1,400 hours or longer, 1,500 hours or longer, 1,600 hours or more, 1,700 hours or more, 1,800 hours or more, 1,900 hours or more, 2,000 hours or more, 2,100 hours or more, 2,200 hours or more, 2,300 hours or more, 2, Patients with a good prognosis were determined to be ≥400 hours, ≥2,500 hours, ≥2,600 hours, ≥2,700 hours, ≥2,800 hours, ≥2,900 hours, or ≥3,000 hours. Patients below that (those who were not determined to have a good prognosis) can be determined to have a poor prognosis.

In a preferred embodiment, whether bFGF is positive or negative can be determined by the ratio of bFGF-positive cells in stromal cells (eg, cancer-associated fibroblasts (CAF)) in cancer tissue. For example, when the ratio of bFGF-positive cells is equal to or greater than a third predetermined value, it can be determined that the subject from which the cancer tissue is derived is bFGF-positive. Moreover, when the ratio of bFGF-positive cells is less than a third predetermined value, it can be determined that the subject from which the cancer tissue is derived is bFGF-negative. Here, whether or not individual cells are bFGF-positive can be determined by immunohistochemical staining using an antibody that binds to bFGF, or histochemical staining techniques such as in situ hybridization to bFGF mRNA. . Typically, these histochemical stains allow cells with significant staining to be positive and cells without significant staining to be negative.

A third predetermined value is a threshold (or cutoff value) for the percentage of bFGF-positive cells to cancer cells. In some aspects, the third predetermined value is 5% or greater, 6% or greater, 7% or greater, 8% or greater, 9% or greater, 10% or greater, 11% or greater, 12% or greater, 13% or greater, 14% 15% or more, 16% or more, 17% or more, 18% or more, 19% or more, 20% or more, 21% or more, 22% or more, 23% or more, 24% or more, 25% or more, 26% or more, It can be a value of 27% or more, 28% or more, 29% or more, or 30% or more. The third predetermined value can be, for example, a value of 5-30%, 5-25%, 5-20%, or 5-15%.

In one preferred embodiment, the cancer patient is a post-cancer resection patient.

In one preferred aspect, the cancer is lung cancer, more preferably lung adenocarcinoma.

In one preferred embodiment, the cancer patient is a post-cancer resection patient and the cancer is lung adenocarcinoma.

<Treatment method of the present disclosure>
According to this disclosure:
(B) a method of treating cancer in a subject with cancer, comprising:
Methods are provided comprising administering a therapeutically effective amount of an anti-angiogenic agent to a subject with a bFGF-negative cancer.

Whether bFGF is negative or not can be determined by the above method (A).

Method (B) above may comprise selecting a subject with a bFGF-negative cancer prior to said administering. The method (B) may further comprise performing the method (A) prior to the administering, wherein the subject with bFGF-negative cancer has a prognostic can be a subject determined to be bad.

Anti-angiogenic agents include, for example, anti-vascular endothelial cell growth factor therapeutic agents (anti-VEGF therapeutic agents). Anti-VEGF therapeutics include, for example, Lucentis™ (ranibizumab), Eylea™ (aflibercept or VEGF Trap), Avastin™ (bevacizumab), and Macugen™ (pegaptanib). be done. Anti-VEGF therapeutics also include, for example, lapatinib (Tikerb™), sunitinib (Sutent™), sorafenib (Nexavar™), and axitinib. Anti-VEGF therapeutic agents can be, for example, antibodies (particularly monoclonal antibodies) or antigen-binding fragments thereof that bind to VEGF, including antibodies that can inhibit activation of VEGF receptors in the presence of VEGF. In certain aspects, an antibody that binds VEGF may bind to a site on VEGF that can inhibit binding of VEGF to a VEGF receptor. The antibody that binds to VEGF includes an antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region containing heavy chain CDRs 1 to 3 of the above antibody and a light chain variable region containing light chain CDRs 1 to 3 of the above antibody. mentioned. Heavy chain CDRs 1-3 and light chain CDRs 1-3 can be identified, for example, using the numbering system according to Kabat (Kabat Sequences of Proteins of Immunological Interest (1987 and 1991, NIH, Bethesda, Md.) or Chothia & Lesk, 1987, J. Mol.Biol.196:901-917; Chothia et al., 1989, Nature 342:878-883)). In one aspect, the regions distinct from the framework regions (especially the amino acid sequences regions of high variability) can be identified as CDRs. Antibodies that compete with any of the above antibodies for binding to VEGF can also be used as anti-angiogenic agents, as well as the above antibodies. Competition can be confirmed using competitive binding assays. For example, when VEGF is immobilized, the first antibody is bound to the immobilized VEGF to form a complex, and then the complex is contacted with a second antibody, the first A first antibody and a second antibody do not compete for binding to VEGF if the antibody does not inhibit the binding of the second antibody. In contrast, if the first antibody inhibits binding of the second antibody, the first and second antibodies compete for binding to VEGF. Compete with each other means that competition is observed even if the first and second antibodies are switched. In certain aspects, antibodies that compete with each other for binding to VEGF with any of the antibodies described above can be used as anti-angiogenic agents, as well as the antibodies described above.

Anti-angiogenic agents include, but are not limited to oral administration, parenteral administration (e.g. intradermal, intramuscular, intraperitoneal, intravenous and subcutaneous), epidural administration, intratumoral administration, mucosal administration (e.g. intranasal and oral routes) and pulmonary administration (eg, an aerosolized compound administered with an inhaler or nebulizer).

A therapeutically effective dose is that dose of active ingredient that confers therapeutic significance. Therapeutic significance may result if the disease slows, stops, alleviates the symptoms of the disease, or cures the disease as compared to the absence of administration. A therapeutically effective amount can be determined by a physician in consideration of the degree of disease progression, sex, age and/or body weight, and can be, for example, 1-100 mg/kg body weight. Administration may be carried out according to a dosing regimen. Dosage regimens may be adjusted to provide the optimum desired response (eg, a therapeutic or prophylactic response). A treatment regimen may be a single dose or may involve cycles of administration.

Anti-angiogenic agents in aqueous formulations, lyophilized formulations, or lyophilized and rehydrated formulations (especially anti-cancer agents or pharmaceutical compositions for use in treating cancer) be. Anti-angiogenic agents can include pharmaceutically acceptable excipients. In some embodiments, the hydrating solution is dextrose and/or saline (eg, about 5% w/v concentration of dextrose and/or about 0.9% w/v concentration of saline).

In some embodiments, the cancer can be lung adenocarcinoma and can be a post-cancer resection subject.

In some embodiments, an anti-angiogenic agent may be administered in combination with lung cancer therapy. Lung cancer therapy may include any or more of chemotherapy, surgery, and radiation therapy. In chemotherapy, subjects are treated with gefitinib (especially if the subject has an EGFR gene mutation), platinum agents (e.g., cisplatin, carboplatin, nedaplatin), antimetabolites (e.g., pyrimidine antimetabolites, e.g., pemetrexed, gemcitabine, TS-1), topoisomerase inhibitors (e.g., irinotecan, nogitecan, etoposide), anticancer antibiotics (e.g., amrubicin), microtubule acting agents (e.g., docetaxel, paclitaxel, vinorelbine), molecular targeted drugs (e.g., bevacizumab, ramucirumab, necitumumab, gefitinib, erlotinib, afatinib, osimertinib, dacomitinib, crizotinib, alectinib, ceritinib, lorlatinib, dabrafenib, trametinib, entrectinib), immune checkpoint inhibitors (e.g., from nivolumab, pemprolizumab, avelumab, atezolimab) can be administered with one or more selected from the following anticancer agents.

According to this disclosure:
(C) a method of treating cancer in a subject with cancer, comprising:
administering a therapeutically effective amount of a chemotherapeutic agent other than an anti-VEGF therapeutic agent to a subject with a bFGF-positive cancer;
A method is provided.

According to this disclosure:
(D) A method of treating cancer in a subject with cancer, comprising:
determining whether the subject has a bFGF-positive cancer;
A therapeutically effective amount of an anti-angiogenic agent is administered to a subject with bFGF-negative cancer, and an anti-angiogenic agent (especially an anti-VEGF therapeutic agent) is administered to a subject with bFGF-positive cancer administering a therapeutically effective amount of a chemotherapeutic agent;
A method is provided.

According to this disclosure:
(E) A method of treating cancer in a subject with cancer, comprising:
Predicting the prognosis of the subject by the method described in (A) above;
administering a therapeutically effective amount of an anti-angiogenic agent to a subject predicted to have a poor prognosis;
A method is provided.

According to this disclosure:
(F) A method of treating cancer in a subject with cancer, comprising:
Predicting the prognosis of the subject by the method described in (A) above;
administering a therapeutically effective amount of a chemotherapeutic agent other than an anti-angiogenic agent (especially an anti-VEGF therapeutic agent) to a subject predicted to have a good prognosis;
A method is provided.

According to this disclosure:
(G) A method of treating cancer in a subject with cancer, comprising:
Predicting the prognosis of the subject by the method described in (A) above;
A therapeutically effective amount of an anti-angiogenic agent is administered to a subject predicted to have a poor prognosis, and an anti-angiogenic agent (especially an anti-VEGF therapeutic agent) is administered to a subject predicted to have a good prognosis administering a therapeutically effective amount of a chemotherapeutic agent;
A method is provided.

In some embodiments, the cancer can be lung adenocarcinoma and can be a post-cancer resection subject.

According to the present disclosure, anti-angiogenic agents (especially anti-VEGF therapeutic agents) are provided for use in the methods of any of (B), (D), (E), and (G) above. . According to the present disclosure, a pharmaceutical composition comprising an anti-angiogenic agent (particularly an anti-VEGF therapeutic agent) for use in any of the methods of (B), (D), (E) and (G) above goods are provided. According to the present disclosure, anti-angiogenic agents (especially anti-VEGF therapeutic agents) in the manufacture of a medicament for use in any of the methods of (B), (D), (E), and (G) above. use is provided. According to the present disclosure there is provided use of an anti-angiogenic agent (particularly an anti-VEGF therapeutic agent) in any of the methods of (B), (D), (E) and (G) above.

According to the present disclosure, there is provided a chemotherapeutic agent, other than an anti-angiogenic agent, for use in any of the methods of (C), (D), (F) and (G) above. According to the present disclosure, a pharmaceutical composition comprising a chemotherapeutic agent other than an anti-angiogenic agent for use in any of the methods of (C), (D), (F), and (G) above is provided. provided. According to the present disclosure, the use of a chemotherapeutic agent other than an anti-angiogenic agent in the manufacture of a medicament for use in any of the methods of (C), (D), (F), and (G) above provided. According to the present disclosure, there is provided use of a chemotherapeutic agent other than an anti-angiogenic agent in any of the methods of (C), (D), (F) and (G) above.

According to the present invention, a kit for use in immunohistochemical staining of lung adenocarcinoma tissue specimens can be provided. The kit of the present invention includes, for example, means for detecting bFGF expression in tissue samples. Means for detecting expression of bFGF include, for example, an antibody that binds to bFGF. Antibodies that bind to bFGF may be labeled or unlabeled. If the antibody that binds to bFGF is not labeled, a secondary antibody that binds to the antibody can be used. The secondary antibody can be labeled, for example. Thus, the kits of the present invention contain a labeled antibody that binds to bFGF, or a labeled or unlabeled antibody that binds to bFGF and a labeled antibody that binds to the antibody. can include a secondary antibody. Moreover, the kit of the present invention may further contain a chromogenic substrate, a fluorescent substrate, or a luminescent substrate suitable for detection. Substrates (chromogenic substrates, fluorescent substrates, and luminescent substrates) and enzymes used in enzyme-antibody methods (eg, peroxidase, glucose oxidase, and alkaline phosphatase) can be used as labels. Substrates include, for example, fluorescent substrates. As a peroxidase, for example, horseradish peroxidase can be used. Horseradish peroxidase produces color, fluorescence or chemiluminescence upon addition of a chromogenic, fluorogenic or luminescent substrate. Therefore, the presence of labeled capture molecules can be detected using chromogenic, fluorescent or chemiluminescent indicators. Chromogenic substrates for horseradish peroxidase include, for example, tetramethylbenzidine (TMB), o-phenylenediamine (OPD), 2,2-azinobis[3-ethylbenzo-thiazoline-6-sulfonic acid (ABTS), and Amplex ( Trademark) Red, which can be used to detect labeled molecules in the presence of hydrogen peroxide. Luminescent substrates for alkaline phosphatase include p-nitrophenyl phosphate (pNPP), 4-methylumbelliferyl phosphate (4-MUP), and AttoPhos™ and can be used for detection of capture molecules. Glucose oxidase oxidizes glucose to generate gluconic acid and hydrogen peroxide. Hydrogen peroxide can be readily detected, for example, using a colorimetric probe for detecting hydrogen peroxide (eg, peroxidase). Hydrogen peroxide can be developed, for example, in the presence of peroxidase and its chromogenic substrate.

Example 1 Analysis of bFGF Expression Using Biopsies of Lung Adenocarcinoma In this example, expression of basic fibroblast growth factor (bFGF) in biopsied or resected tissues from patients with lung adenocarcinoma. examined.

Of the 182 cases resected from October 2010 to September 2017 at a city general hospital, 102 cases of lung adenocarcinoma for which immunostaining and clinicopathological examination were possible were used as specimens. The mean age of patients was 70.5 years, and included patients from 43 to 89 years of age. Patient details were as set forth in Table 1 below.

In Table 1, DM represents diabetes, CEA represents expression of CEA markers, and EGFR represents epidermal growth factor receptor. EGFR mutations are any of the driver mutations G719X, a deletion in exon 19 (del19), S768I, an insertion in exon 20, T790M, L858R, and L861Q.

Tissue sections were prepared from biopsies or tissue donations according to a standard method and subjected to immunohistochemical staining. An anti-FGF2 antibody (Abcam; 1/500) was used as the primary antibody. As a secondary antibody, the rabbit anti-mouse IgG antibody included in the BOND Polymer Refine Detection (Leica Biosystems) kit was used according to the manufacturer's instructions. Staining was then performed using a horseradish peroxidase (HRP)-labeled anti-rabbit IgG antibody. Staining was performed with an automatic immunostainer BOND-MAX (Leica Biosystems). Stained images were evaluated using a virtual slide (equipment used: Nano Zoomer-XR (Hamamatsu Photonics Co., Ltd.), conditions: 20x mode, Single Layer).

Immunohistochemical staining showed no obvious staining in the tumor. However, bFGF-positive cells were found in tumor stromal cells (cancer-associated fibroblasts; CAF). FIG. 1 shows the results of immunohistochemical staining of representative lung adenocarcinoma tissues.

Therefore, when 10% or more of the stromal cells in the tumor were evaluated as bFGF-positive, 13 of the 102 cases were bFGF-positive, and 79 of the 79 cases were bFGF-positive. There were 9 cases. In addition, 9 of the 71 stage I cases were bFGF-positive.

As shown in Table 2, no clear correlation was observed between the bFGF-positive frequency and age, sex, smoking status, CEA, EGF receptor mutation, and stage.

In this case of 102 cases, stage II-IV significantly recurred compared to stage I without lymph node metastasis (p = 0.0005) (Fig. 2), and stage III-IV compared to stage I-II with localized lung cancer. relapsed significantly (p=0.0009) (Fig. 3).

In this 102 cases, stage II-IV had a significantly worse prognosis than stage I without lymph node metastasis (p<0.0001) (Fig. 4), and stage III compared to stage I-II with localized lung cancer. -IV had a significantly worse prognosis (p<0.0001) (Figure 5).

In all cases (n = 102), FGF2-positive patients are less likely to recur (Fig. 6), and even in stage I-II (n = 79), which is localized lung cancer, FGF2-positive patients are less likely to recur (DFI: Disease Free intervals). (FIG. 7), FGF2-positive patients were less likely to recur even in stage I (n=71) without lymph node metastasis (FIG. 8).

In all cases (n = 102), FGF2-positive patients tend to have a favorable prognosis (OS: overall survivals) (Fig. 9), and even in stage I-II (n = 79), which is localized lung cancer, FGF2-positive patients have a favorable prognosis. (Fig. 10), and FGF2-positive patients had a good prognosis even in stage I (n = 71) without lymph node metastasis (Fig. 11).

Thus, in biopsies of lung adenocarcinoma, positive bFGF expression in stromal cells indicated a good prognosis, and negative results indicated a poor prognosis.

Claims (11)

A method for predicting the prognosis of a patient with lung adenocarcinoma, comprising:
detecting expression of basic fibroblast growth factor (bFGF) in lung adenocarcinoma tissue obtained from a patient with lung adenocarcinoma, wherein positive bFGF indicates a favorable prognosis or A method in which the possibility is indicated and, when negative, the prognosis is or is indicated to be poor. 2. The method of claim 1, wherein the cancer patient is a post-cancer resection patient. 3. The method of claim 1 or 2, wherein the level of expression is determined based on the ratio of positive cells to cancer cells. A method according to any one of claims 1 to 3, wherein said expression is protein expression. The method according to any one of claims 1 to 4, wherein the prognosis is at least one selected from the group consisting of recurrence rate and survival rate. The method of any one of claims 1-5, wherein the lung adenocarcinoma is an early stage cancer selected from the group consisting of stages I and II. A pharmaceutical composition for use in treating a patient with lung adenocarcinoma, comprising:
comprising a therapeutically effective amount of an anti-vascular endothelial growth factor (VEGF) agent;
A pharmaceutical composition, wherein the lung adenocarcinoma patient is a patient evaluated to have a poor prognosis by the method according to any one of claims 1 to 6. 8. The pharmaceutical composition according to claim 7, wherein the anti-VEGF drug is bevacizumab. A method of treating cancer in a subject with cancer, comprising:
predicting the prognosis of the subject by the method of any one of claims 1 to 6;
A pharmaceutical composition comprising an anti-angiogenic agent for use in a method comprising administering a therapeutically effective amount of the anti-angiogenic agent to a subject predicted to have a poor prognosis. A pharmaceutical composition for use in treating a patient with lung adenocarcinoma, comprising:
including cancer chemotherapeutic agents other than anti-vascular endothelial growth factor (VEGF) agents;
A pharmaceutical composition, wherein the lung adenocarcinoma patient is a patient evaluated to have a good prognosis by the method according to any one of claims 1 to 6. A method of treating cancer in a subject with cancer, comprising:
predicting the prognosis of the subject by the method of any one of claims 1 to 6;
The cancer for use in a method comprising administering to a subject predicted to have a good prognosis a therapeutically effective amount of a cancer chemotherapeutic agent other than an anti-vascular endothelial growth factor (VEGF) drug A pharmaceutical composition comprising a chemotherapeutic agent of

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