Biomarkers for the treatment of interstitial lung disease

Abstract

Providing biomarkers for the treatment of interstitial lung disease. [Solution] This disclosure provides biomarkers associated with caveolin-1 peptide therapy in subjects with interstitial lung disease. In particular, this disclosure describes biomarkers such as MYDGF, soluble RAGE, pSMAD2 / 3, and PDGFRβ associated with caveolin-1 peptide therapy in subjects with idiopathic pulmonary fibrosis. These biomarkers can be used to determine the efficacy, monitoring, and optimal dosage of caveolin-1 peptide therapy in subjects with interstitial lung disease.

Description

Technical Field

[0001] Cross-Reference to Related Applications

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 153,565 filed on February 25, 2021, and U.S. Provisional Patent Application No. 63 / 270,867 filed on October 22, 2021, the entire contents of which are incorporated herein by reference in their entirety.

[0002] Technical Field

[0002] The present disclosure relates generally to the fields of medicine, lung diseases, and protein biology. In particular, the present disclosure relates to biomarkers associated with caveolin-1 peptide therapy for the treatment of interstitial lung diseases such as idiopathic pulmonary fibrosis.

[0003] Sequence Listing

[0003] The Sequence Listing associated with this application is provided in text format in lieu of a paper copy, and is incorporated herein by reference. The name of the text file containing the Sequence Listing is LUTX_020_02WO_SeqList_ST25.txt. The text file is about 31KB in size, created on February 25, 2022, and transmitted electronically via EFS-Web.

Background Art

[0004] Background

[0004] Interstitial lung disease (ILD) is a group of more than 200 respiratory diseases that cause fibrosis and inflammation of the tissues and spaces around the air sacs of the lungs. Idiopathic pulmonary fibrosis (IPF) is a type of ILD, which is estimated to affect more than 200,000 people in the United States, and 1 in every 200 adults over 60 years old. IPF is characterized by progressive interstitial fibrosis of the lung parenchyma, which leads to loss of lung function and death from respiratory failure in most patients. The median survival time from diagnosis is 2 to 3 years (Raghu et al., Am J Respir Crit Care Med 183:788-824 (2011)). The etiology of IPF as well as the major molecular and pathophysiological factors remain unknown.

[0005]

[0005] Pirfenidone and nintedanib are approved by the FDA in the United States as treatments for IPF. Pirfenidone suppresses transforming growth factor beta-1 (TGF-β1)-stimulated collagen synthesis and downregulates pro-inflammatory cytokines. Nintedanib is a broad-spectrum tyrosine kinase inhibitor with anti-fibrotic properties. Both compounds have been shown to slow disease progression in patients with IPF, but neither treatment is known to be curative, and many patients experience side effects that reduce their quality of life. Lung transplantation is one of the treatment options for IPF, but it is expensive and associated with a significant mortality rate, as only about 50% of lung transplant recipients survive to 5 years.

[0006]

[0006] Caveolin-1 is an intrinsic membrane protein that plays a homeostatic role in the fibrotic process by being involved in a series of major regulatory pathways, including TGF-β signaling (Shihata et al., Front.Pharmacol.2017; 8:567; and Gvaramia et al., Matrix Biol, 2013:32(6):307-315). Endogenous caveolin-1 was constitutively suppressed in bleomycin-induced animal models of IPF and in fibrotic lungs in IPF patients (Wang et al., J Exp Med, 2006; 203(13):2895-2906; Sanders et al., PLoS One, 2015; 10(2), e0116995; and Sanders et al., Am J Respir Cell Mol Biol, 2017; 56(1):50-61). Full-length caveolin-1 protein may not be a very desirable drug candidate due to concerns inherent to protein drugs, including stability, delivery, cost, and autoimmunogenicity. Therefore, fragments of caveolin-1, such as caveolin scaffold domain peptides (CSPs), have been studied and found to be effective alternatives to full-length caveolin-1 protein. In particular, the 20-mer form of CSP has been shown to prevent, limit, or reverse fibrosis in animal models, and a 7-amino acid fragment of CSP, called CSP-7, was sufficiently effective in promoting fibrosis reduction both in vitro and in vivo (Marudamuthu et al., Sci Transl Med, 2019; 11(522), eaat2848). As a result, CSPs have been identified as promising therapeutic agents for the treatment of fibrotic diseases such as IPF.

[0007]

[0007] No biomarkers associated with caveolin-1 peptide therapy in patients with IPF have yet been identified. These biomarkers may be used to determine or predict the efficacy of caveolin-1 peptide therapy. These biomarkers may also be used to determine the optimal dose of caveolin-1 peptide therapy in patients with IPF.

[0008]

[0008] Therefore, in the art, it is necessary to identify biomarkers associated with caveolin-1 peptide therapy in patients with interstitial lung diseases such as IPF. [Overview of the Initiative] [Means for solving the problem]

[0009] Summary of Disclosure

[0009] This disclosure provides a method for treating a subject with caveolin-1 peptide therapy, wherein the subject suffers from fibrosis, and the method comprises (a) obtaining a biological sample from the subject or the step of obtaining one; (b) treating cells of the biological sample with caveolin-1 peptide or a derivative thereof or the step of having treated cells with caveolin-1 peptide or a derivative thereof; (c) measuring the expression level of a biomarker in the cells; and (d) comparing the expression level of the biomarker with that of a control sample; the biomarkers include bone marrow-derived growth factor (MYDGF), soluble RAGE, and phosphorylated Mothers Against • Decapentaplesic homolog 2 / 3 (pSMAD2 / 3), platelet-derived growth factor receptor beta (PDGFRβ), galectin-7 (LGALS7), interleukin-11 (IL-11), matrix metalloproteinase-2 (MMP-2), chemokine ligand 7 (CXCL-7), soluble CD163, phosphorylated mTOR (p-mTOR), phosphorylated PDGFRβ (pPDGFRβ), prolifin (PROF1), calmodulin 2 (CALM2), calreticulin (CALR), peptidyl-prolyl cis-trans isomerase A (PPIA), or eukaryotic translation initiation factor 5A (EIF5A1);

[0010] If the expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 are increased, administer caveolin-1 peptide or its derivatives to the target; or

[0011] If the expression levels of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, soluble CD163, or pSMAD2 / 3 are decreased, administer caveolin-1 peptide or its derivatives.

[0010]

[0012] This disclosure also provides a method for identifying changes in the expression levels of biomarkers associated with caveolin-1 peptide therapy; the method comprises (a) treating target cells having interstitial lung disease with caveolin-1 peptide or a derivative thereof; (b) measuring the expression level of the biomarker in the cells; and (c) comparing the expression level of the biomarker with a control sample; wherein the caveolin-1 peptide or a derivative thereof modulates the expression level of the biomarker in the cells; the biomarkers include bone marrow-derived growth factor (MYDGF), soluble RAGE, and phosphorylated mother's age. These include ionst-decapentaplesic homolog 2 / 3 (pSMAD2 / 3), platelet-derived growth factor receptor beta (PDGFRβ), phosphorylated PDGFRβ (pPDGFRβ), phosphorylated mTOR (p-mTOR), galectin-7 (LGALS7), interleukin-11 (IL-11), matrix metalloproteinase-2 (MMP-2), chemokine ligand 7 (CXCL-7), soluble CD163 (sCD163), prolifin (PROF1), calmodulin 2 (CALM2), calreticulin (CALR), peptidyl-prolyl cis-trans isomerase A (PPIA), or eukaryotic translation initiation factor 5A (EIF5A1). In some embodiments, steps (a) to (c) of this method are repeated once or more times.

[0011]

[0013] In some embodiments, the control sample is cells obtained from subjects with interstitial lung disease prior to treatment with caveolin-1 peptide or its derivatives. In some embodiments, the control sample is cells obtained from subjects or a population of subjects with interstitial lung disease. In some embodiments, the control sample is cells obtained from healthy subjects.

[0012]

[0014] In some embodiments, the cells are fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal-like cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphoid cells, T cells, monocytes, NK cells, dendritic cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, the cells are fibroblasts. In some embodiments, the cells are basal-like cells. In some embodiments, the cells are treated with caveolin-1 peptide or its derivatives ex vivo or in vitro. In some embodiments, the cells are obtained from subjects having interstitial lung disease.

[0013]

[0015] In some embodiments, the caveolin-1 peptide or its derivative is FTFTVT (SEQ ID NO: 3). In some embodiments, the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTVTKGSaaNH2 (SEQ ID NO: 4).

[0014]

[0016] In some embodiments, the subject is a human.

[0015]

[0017] In some embodiments, ILD is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar injury, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced ILD, or occupational ILD. In some embodiments, ILD is idiopathic pulmonary fibrosis.

[0016]

[0018] In some embodiments, the expression level of the biomarker is measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

[0017]

[0019] In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 are associated with caveolin-1 therapy. In some embodiments, decreased expression levels of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 are associated with caveolin-1 therapy. In some embodiments, MYDGF expression levels increase compared to a control sample. In some embodiments, MYDGF expression levels increase by approximately 1.5 times, approximately 2 times, approximately 3 times, approximately 4 times, approximately 5 times, or more, compared to a control sample. In some embodiments, soluble RAGE expression levels increase compared to a control sample. In some embodiments, soluble RAGE expression levels increase by approximately 1.5 times, approximately 2 times, approximately 3 times, approximately 4 times, approximately 5 times, or more, compared to a control sample. In some embodiments, the expression level of pSMAD2 / 3 is reduced compared to the control sample. In some embodiments, the expression level of pSMAD2 / 3 is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more, compared to the control sample. In some embodiments, the expression level of PDGFRβ is reduced compared to the control sample. In some embodiments, the expression level of PDGFRβ is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more, compared to the control sample.

[0018]

[0020] In some embodiments, the method further includes measuring the expression level of an internal control, where the expression level of the internal control is unaffected by caveolin-1 peptide or its derivatives.

[0019]

[0021] In some embodiments, the present disclosure provides a method for predicting or determining the efficacy of a therapeutic activator, the method comprising: (a) treating target cells having interstitial lung disease with the therapeutic activator; (b) measuring the expression level of a biomarker in the target cells, wherein the biomarker is MYDGF, soluble RAGE, pSMAD2 / 3, PDGFRβ, pPDGFRβ, p-mTOR, LGALS7, IL-11, MMP-2, CXCL7, sCD163, PROF1, CALM2, CALR, PPIA, or EIF5A1; and (c) comparing the expression level of the biomarker with that of a control sample. In some embodiments, steps (a) to (c) of the method are repeated once or more times.

[0020]

[0022] In some embodiments, the control sample is cells obtained from subjects with interstitial lung disease before treatment with the therapeutic agent. In some embodiments, the control sample is cells obtained from subjects or a population of subjects with interstitial lung disease. In some embodiments, the control sample is cells obtained from healthy subjects.

[0021]

[0023] In some embodiments, the cells are fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal-like cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphoid cells, T cells, monocytes, NK cells, dendritic cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, the cells are fibroblasts. In some embodiments, the cells are basal-like cells. In some embodiments, the cells are treated with a therapeutic activator ex vivo or in vitro. In some embodiments, the cells are obtained from subjects having interstitial lung disease.

[0022]

[0024] In some embodiments, the therapeutically active agent is a small molecule. In some embodiments, the therapeutically active agent is a biological agent. In some embodiments, the biological agent is a caveolin-1 peptide or a derivative thereof. In some embodiments, the caveolin-1 peptide or derivative thereof is FTTFTVT (SEQ ID NO: 3). In some embodiments, the caveolin-1 peptide or derivative thereof is Ac-aaEGKASFTTFTVTKGSaaNH2 (SEQ ID NO: 4).

[0023]

[0025] In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar damage, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0024]

[0026] In some embodiments, the expression level of the biomarker is measured by two-dimensional gel electrophoresis, Western blot, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

[0025]

[0027] In some embodiments, an increase in the expression level of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 indicates a favorable response to a therapeutically active agent. In some embodiments, a decrease in the expression level of PDGFRβ, pPDGFRβ, p-mTOR, pSMAD2 / 3, IL-11, MMP-2, CXCL7, sCD163, or CALR indicates a favorable response to a therapeutically active agent. In some embodiments, the expression level of MYDGF is increased compared to a control sample. In some embodiments, the expression level of MYDGF is increased by about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, or more, compared to a control sample. In some embodiments, the expression level of soluble RAGE is increased compared to a control sample. In some embodiments, the expression level of soluble RAGE is increased by about 1.5-fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, or more, compared to a control sample. In some embodiments, the expression level of pSMAD2 / 3 is decreased compared to a control sample. In some embodiments, the expression level of pSMAD2 / 3 is decreased by about 10%, about 25%, about 50%, about 75%, about 90%, or more, compared to a control sample. In some embodiments, the expression level of PDGFRβ is decreased compared to a control sample. In some embodiments, the expression level of PDGFRβ is decreased by about 10%, about 25%, about 50%, about 75%, about 90%, or more, compared to a control sample.

[0026]

[0028] In some embodiments, the method further comprises measuring the expression level of an internal control, wherein the expression level of the internal control is not affected by the therapeutically active agent.

[0027]

[0029] In some embodiments, the Disclosure provides a method for predicting or determining the efficacy of a caveolin-1 peptide or a derivative thereof, the method comprising: (a) treating a biological sample from a subject having fibrosis with a caveolin-1 peptide or a derivative thereof; (b) measuring the expression level of a biomarker in the biological sample from the subject, wherein the biomarker is MYDGF, soluble RAGE, pSMAD2 / 3, PDGFRβ, pPDGFRβ, p-mTOR, LGALS7, IL-11, MMP-2, CXCL7, sCD163, PROF1, CALM2, CALR, PPIA, or IF5A1; and (c) comparing the expression level of the biomarker with that of a control sample. In some embodiments, steps (a) to (c) of the method are repeated once or more times.

[0028]

[0030] In some embodiments, the control sample is a biological sample obtained from a subject with fibrosis prior to treatment with caveolin-1 peptide or its derivatives. In some embodiments, the control sample is a biological sample obtained from a subject or population of subjects with fibrosis. In some embodiments, the control sample is a biological sample obtained from a healthy subject.

[0029]

[0031] In some embodiments, the biological sample is bronchoalveolar lavage fluid (BALF), lung tissue, fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphoid cells, T cells, monocytes, NK cells, dendritic cells, or PBMCs. In some embodiments, the biological sample is fibroblasts. In some embodiments, the biological sample is basal-like cells. In some embodiments, the biological sample is lung tissue. In some embodiments, the biological sample is treated with caveolin-1 peptide or its derivatives ex vivo or in vitro. In some embodiments, the biological sample is obtained from a subject with fibrosis.

[0030]

[0032] In some embodiments, the caveolin-1 peptide or its derivative is FTFTVT (SEQ ID NO: 3). In some embodiments, the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTVTKGSaaNH2 (SEQ ID NO: 4).

[0031]

[0033] In some embodiments, fibrosis is interstitial lung disease, hepatic fibrosis, renal fibrosis, dermatofibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataracts, retina and vitreoretinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloids and hypertrophic scars, peritoneal fibrous disease, chronic obstructive pulmonary disease, postoperative fibroids, diabetic nephropathy, gynecological cancers, myeloproliferative syndromes, myeloid leukemia, myelodysplastic syndromes, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome.

[0032]

[0034] In some embodiments, interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar injury, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease. In some embodiments, interstitial lung disease is idiopathic pulmonary fibrosis.

[0033]

[0035] In some embodiments, the expression level of the biomarker is measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

[0034]

[0036] In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, and soluble RAGE indicate a favorable response to caveolin-1 peptides or their derivatives. In some embodiments, decreased expression levels of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicate a favorable response to caveolin-1 peptides or their derivatives. In some embodiments, the expression level of MYDGF increases compared to the control sample. In some embodiments, the expression level of MYDGF increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more, compared to the control sample. In some embodiments, the expression level of soluble RAGE increases compared to the control sample. In some embodiments, the expression level of soluble RAGE increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more, compared to the control sample. In some embodiments, the expression level of pSMAD2 / 3 is reduced compared to the control sample. In some embodiments, the expression level of pSMAD2 / 3 is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more, compared to the control sample. In some embodiments, the expression level of PDGFRβ is reduced compared to the control sample. In some embodiments, the expression level of PDGFRβ is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more, compared to the control sample.

[0035]

[0037] In some embodiments, the method further includes measuring the expression level of an internal control, where the expression level of the internal control is unaffected by caveolin-1 peptide or its derivatives.

[0036]

[0038] In some embodiments, the Disclosure provides a method for predicting or determining the efficacy of a caveolin-1 peptide or derivative thereof in a subject having fibrosis, the method comprising: (a) obtaining a first biological sample from the subject before treatment with the caveolin-1 peptide or derivative thereof; (b) administering the caveolin-1 peptide or derivative thereof to the subject; (c) obtaining a second biological sample from the subject after treatment with the caveolin-1 peptide or derivative thereof; (d) measuring the expression levels of a biomarker in the first and second biological samples, wherein the biomarker is MYDGF, soluble RAGE, pSMAD2 / 3, PDGFRβ, pPDGFRβ, p-mTOR, LGALS7, IL-11, MMP-2, CXCL7, sCD163, PROF1, CALM2, CALR, PPIA, or IF5A1; and (e) comparing the expression level of the biomarker in the first biological sample with the expression level of the biomarker in the second biological sample. In some embodiments, step (a) of the method further includes determining the expression level of a biomarker in a first biological sample compared to a control sample before treatment with the caveolin-1 peptide or a derivative thereof. In some embodiments, the expression level of the biomarker in the first biological sample from step (a) is used to determine the optimal dose of the caveolin-1 peptide or a derivative thereof to be administered in step (b).

[0037]

[0039] In some embodiments, the control sample is a biological sample obtained from a subject or population of subjects with fibrosis.

[0038]

[0040] In some embodiments, the biological sample is serum, plasma, BALF, lung tissue, fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphoid cells, T cells, monocytes, NK cells, dendritic cells, or PBMCs. In some embodiments, the biological sample is serum. In some embodiments, the biological sample is plasma. In some embodiments, the biological sample is fibroblasts, basal-like cells, or PBMCs.

[0039]

[0041] In some embodiments, the caveolin-1 peptide or its derivative is FTFTVT (SEQ ID NO: 3). In some embodiments, the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTVTKGSaaNH2 (SEQ ID NO: 4).

[0040]

[0042] In some embodiments, fibrosis is interstitial lung disease, hepatic fibrosis, renal fibrosis, dermatofibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataracts, retina and vitreoretinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloids and hypertrophic scars, peritoneal fibrous disease, chronic obstructive pulmonary disease, postoperative fibroids, diabetic nephropathy, gynecological cancers, myeloproliferative syndromes, myeloid leukemia, myelodysplastic syndromes, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome.

[0041]

[0043] In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis, familial pulmonary fibrosis, idiopathic nonspecific interstitial pneumonia, conventional interstitial lung disease, idiopathic organizing pneumonia, or sarcoidosis. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0042]

[0044] In some embodiments, the expression levels of biomarkers in the first and second biological samples are measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

[0043]

[0045] In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 indicate a favorable response to caveolin-1 peptides or their derivatives. In some embodiments, decreased expression levels of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicate a favorable response to caveolin-1 peptides or their derivatives.

[0044]

[0046] In some embodiments, the caveolin-1 peptide or its derivatives are administered to subjects at doses ranging from approximately 0.01 mg / kg to approximately 250 mg / kg. In some embodiments, the caveolin-1 peptide or its derivatives are administered to subjects at doses ranging from approximately 0.05 mg / kg to approximately 50 mg / kg.

[0045]

[0047] In some embodiments, caveolin-1 peptide or its derivatives are administered to the subject by inhalation, intravenous, subcutaneous, oral, intraperitoneal, sublingual, buccal, or intramuscular means.

[0046]

[0048] In some embodiments, the method involves administering a caveolin-1 peptide or a derivative thereof to a subject once daily, once weekly, twice weekly, three times weekly, five times weekly, once every two weeks, or once monthly.

[0047]

[0049] In some embodiments, the second biological sample is obtained from the subject at 1 hour, 3 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 1 week, 2 weeks, 3 weeks, 1 month, 6 months, or 1 year after administration of caveolin-1 peptide or its derivative.

[0048]

[0050] In some embodiments, the method further comprises obtaining one or more additional biological samples after administration of caveolin-1 peptide or a derivative thereof. In some embodiments, the expression level of the biomarker in one or more additional biological samples is increased compared to the expression level of the biomarker in the first biological sample; the increase in the expression level of the biomarker in one or more additional biological samples indicates a favorable response to caveolin-1 peptide or a derivative thereof. In some embodiments, the expression level of the biomarker in one or more additional biological samples is decreased compared to the expression level of the biomarker in the first biological sample; the decrease in the expression level of the biomarker in one or more additional biological samples indicates a favorable response to caveolin-1 peptide or a derivative thereof.

[0049]

[0051] In some embodiments, the expression level of MYDGF in the second biological sample increases compared to the first biological sample. In some embodiments, the expression level of MYDGF in the second biological sample increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more, compared to the first biological sample. In some embodiments, the expression level of soluble RAGE in the second biological sample increases compared to the first biological sample. In some embodiments, the expression level of soluble RAGE in the second biological sample increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more, compared to the first biological sample. In some embodiments, the expression level of pSMAD2 / 3 decreases compared to the control sample. In some embodiments, the expression level of pSMAD2 / 3 decreases by about 10%, about 25%, about 50%, about 75%, about 90%, or more, compared to the control sample. In some embodiments, the expression level of PDGFRβ in the second biological sample is reduced compared to the first biological sample. In some embodiments, the expression level of PDGFRβ in the second biological sample is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the first biological sample.

[0050]

[0052] In some embodiments, the expression level of a biomarker in a second biological sample is used to determine the optimal dose of caveolin-1 peptide or its derivative. [Brief explanation of the drawing]

[0051] Brief explanation of the drawing [Figure 1A]

[0053] Figure 1A shows MYDGF expression in epithelial cells from IPF patients. Epithelial cells were treated with 1% DMSO vehicle, CSP-7 (10 μM), Var55 (10 μM), control peptide 1 (CP1, 10 μM), control peptide 2 (CP2, 10 μM), pirfenidone (500 μM), or nintedanib (100 nM). MYDGF expression in the epithelial cell supernatant was measured by ELISA. [Figure 1B]

[0053] Figure 1B shows MYDGF expression in epithelial cells from IPF patients. Epithelial cells were treated with 1% DMSO vehicle, CSP-7 (10 μM), Var55 (10 μM), control peptide 1 (CP1, 10 μM), control peptide 2 (CP2, 10 μM), pirfenidone (500 μM), or nintedanib (100 nM). MYDGF expression in the epithelial cell supernatant was measured by ELISA. [Figure 2]

[0054] Figure 2 shows a schematic diagram of ex vivo testing of therapeutic agents in human IPF lung tissue. Fresh lung tissue is obtained from human subjects with IPF, the lung core is perforated, and the tissue is transferred to a Krumdieck slicer. Precision-cut lung sections (PCLS) are kept alive for up to one month with human serum and culture medium, and the therapeutic agent, such as CSP-7, is directly tested against diseased organs composed of multiple cell types. [Figure 3A]

[0055] Figure 3A shows soluble RAGE expression in the supernatant of IPF precision lung sections (PCLS). PCLS were either untreated (UT) or treated with CSP-7 (10 μM or 100 μM), control peptide (CP, 100 μM), TGFβ (2 ng / mL), or pirfenidone (Pirf, 500 μM) and nintedanib (Nin, 100 nM). Soluble RAGE expression was measured in the supernatant of IPF PCLS on days 2 and 4 after ELISA treatment. [Figure 3B]

[0055] Figure 3B shows soluble RAGE expression in the supernatant of IPF precision lung sections (PCLS). PCLS were either untreated (UT) or treated with CSP-7 (10 μM or 100 μM), control peptide (CP, 100 μM), TGFβ (2 ng / mL), or pirfenidone (Pirf, 500 μM) and nintedanib (Nin, 100 nM). Soluble RAGE expression was measured in the supernatant of IPF PCLS on days 2 and 4 after ELISA treatment. [Figure 3C]

[0055] Figure 3C shows the effects of different concentrations of CSP-7 (0.001 μM to 100 μM) on soluble RAGE expression in IPF PCLS. Soluble RAGE expression was measured in the supernatant of IPF PCLS on days 2 and 4 after ELISA treatment. [Figure 4A]

[0056] Figure 4A shows lactate dehydrogenase (LDH) activity in precision lung sections (PCLS) from end-stage IPF. PCLS were either untreated or treated with CSP-7 (10 μM or 100 μM), control peptide (CP, 100 μM), TGFβ (2 ng / mL), or pirfenidone (Pirf, 500 μM) and nintedanib (Nin, 100 nM). LDH activity in the supernatant of IPF PCLS was measured on day 2 (left panel) or day 4 (right panel) after treatment. [Figure 4B]

[0056] Figure 4B shows the LDH activity in end-stage IPFPCLS that were untreated (UT) or treated with different concentrations of CSP-7 (0.001 μM to 100 μM). The LDH activity in the supernatant of IPF PCLS was measured on day 2 (left panel) or day 4 (right panel) after treatment. [Figure 5]

[0057] Figure 5 shows soluble RAGE expression in the plasma of healthy subjects on days 1 and 13 after CSP-7 inhalation. Soluble RAGE expression was measured by ELISA. [Figure 6A]

[0058] Figure 6A shows PDGFRB expression in precision lung sections (PCLS) from three biological replicas of end-stage IPF. PCLS were treated with CSP-7 (10 μM or 100 μM), control peptide (CP, 100 μM), TGFβ (2 ng / mL), pirfenidone (500 μM), or nintedanib (100 nM). Protein expression was measured by Western blotting. Beta-actin was used as an endogenous control. [Figure 6B]

[0058] Figure 6B shows relative PDGFRB protein expression in precision lung sections (PCLS) of end-stage IPF. PCLS were treated with CSP-7 (10 μM or 100 μM), control peptide (CP, 100 μM), TGFβ (2 ng / mL), or pirfenidone (Pirf, 500 μM) and nintedanib (Nin, 100 nM). Relative protein expression was determined using the data shown in Figure 6A. *P ≤ 0.05 and **P ≤ 0.01. [Figure 7A]

[0059] Figure 7A shows the phosphorylation of pSMAD2 / 3 in IPF fibroblasts. IPF fibroblasts were treated with DMEM, 1% DMSO vehicle, CSP-7 (10 μM), Var55 (10 μM), pirfenidone (500 μM), or nintedanib (100 nM). SMAD2 / 3 phosphorylation was determined by Western blotting 24 hours after treatment. Phosphorylated SMAD2 / 3 protein levels were determined by normalizing to total SMAD2 / 3 protein. [Figure 7B]

[0059] Figure 7B shows the phosphorylation of pSMAD2 / 3 in IPF epithelial cells. IPF epithelial cells were treated with DMEM (control), Var55 (100 μM), nintedanib (100 nM), control peptide 1 (CP1, 100 μM), or control peptide 2 (CP2, 100 μM). SMAD2 / 3 phosphorylation was determined by Western blotting 24 hours after treatment. Phosphorylated SMAD2 / 3 protein levels were determined by normalizing to total SMAD2 / 3 protein. [Figure 8]

[0060] Figure 8 shows galectin-7 in basal-like cells and fibroblasts derived from normal healthy subjects or subjects with idiopathic pulmonary fibrosis (IPF). Cells were treated with DMSO vehicle, CSP-7 (10 μM), Var55 (10 μM), control peptide 1 (CP1, 10 μM), control peptide 2 (CP2, 10 μM), pirfenidone (500 μM), or nintedanib (100 nM). [Figure 9]

[0061] Figure 9 shows collagen in whole lung homogenates of aged mice treated with saline (SAL) or bleomycin (BLM) after administration of a control peptide (CP) or CSP-7 by dry powder inhalation (DPI). ****P ≤ 0.0001. [Figure 10A]

[0062] Figure 10A shows total SMAD (tSMAD, upper panel) and phosphorylated SMAD (pSMAD, lower panel) in whole lung homogenates of aged mice that were either untreated or treated with bleomycin (BLM) by dry powder inhalation of a control peptide (CP) or CSP-7, or by intraperitoneal injection (IP) of CSP-7. [Figure 10B]

[0063] Figure 10B shows galectin-7 in whole lung homogenates of bleomycin (BLM)-treated aged mice, either untreated or administered by dry powder inhalation of a control peptide (CP) or CSP-7, or by intraperitoneal injection (IP) of CSP-7. [Figure 11A-1]

[0064] Figure 11A shows the phosphorylation of EGFR (p-EGFR), SRC (p-SRC), MEK1 / 2 (p-MEK1 / 2), and Raptor (p-Raptor) in untreated or treated IPF (upper panel) or normal lung fibroblasts (lower panel) with DMSO, control peptide (CP, 100 μM), or CSP-7 (10 μM). [Figure 11A-2]

[0064] Figure 11A shows the phosphorylation of EGFR (p-EGFR), SRC (p-SRC), MEK1 / 2 (p-MEK1 / 2), and Raptor (p-Raptor) in IPF (upper panel) or normal lung fibroblasts (lower panel) that are either untreated or treated with DMSO, control peptide (CP, 100 μM), or CSP-7 (10 μM). [Figure 11B]

[0065] Figure 11B shows the phosphorylation of PDGFRβ (p-PDGFRβ) and mTOR (p-mTOR) in untreated or treated IPF with DMSO, control peptide (CP, 100 μM), or CSP-7 (10 μM). [Figure 11C]

[0066] Figure 11C provides an overview of reverse-phase protein arrays evaluating receptor tyrosine kinase (RTK) signaling, metabolic signaling, infiltration-related markers, and histone deacetylases (HDACs). p-values ​​were calculated by comparing IPF treated with a control peptide (CP) or CSP-7. [Figure 12A]

[0067] Figure 12A shows the IL-11 expression levels in bronchoalveolar lavage fluid (BALF) from mice treated with physiological saline, bleomycin (BLM), BLM and CSP-7, BLM and control peptide (CP), or BLM and CSP-7 administered by intraperitoneal (IP) injection. In the prophylactic group, mice were administered CP or CSP-7 three times daily prior to BLM administration. [Figure 12B]

[0068] Figure 12B shows the IL-11 expression levels in bronchoalveolar lavage fluid (BALF) from mice treated with physiological saline, bleomycin (BLM), BLM and CSP-7, BLM and control peptide (CP), or BLM and CSP-7 administered by intraperitoneal (IP) injection. The treatment group mice received CP or CSP-7 three times daily after BLM administration. [Figure 13A]

[0069] Figure 13A shows soluble CD163 (sCD163) expression in the supernatant of human lung macrophages isolated from subjects with idiopathic pulmonary fibrosis (IPF). IPF macrophages were either left untreated or treated with 0.1 μM or 1 μM CSP-7 for 24 hours prior to sCD163 analysis. Each line represents the biological replication of lung macrophages from individual subjects with IPF. [Figure 13B]

[0070] Figure 13B shows MMP-2 expression in the supernatant of human lung macrophages isolated from subjects with idiopathic pulmonary fibrosis (IPF). IPF lung macrophages were either left untreated or treated with 0.1 μM or 1 μM CSP-7 for 24 hours prior to MMP-2 analysis. Each line represents the biological replication of lung macrophages from individual subjects with IPF. [Figure 14]

[0071] Figure 14 shows CXCL7 expression in the supernatant of human lung macrophages isolated from healthy controls or subjects with idiopathic pulmonary fibrosis (IPF). Lung macrophages were either left untreated or treated with Var55. Each line represents the biological replication of lung macrophages from individual controls (triangles) or individuals with IPF (circles). [Modes for carrying out the invention]

[0052] Detailed explanation definition

[0072] Unless otherwise defined, all technical terms, notations, and other scientific terms used herein are intended to have meanings commonly understood by those skilled in the art relating to the present invention. Where applicable, terms having commonly understood meanings are defined herein for clarity and / or for easy reference, and the inclusion of such definitions herein should not necessarily be interpreted as representing a substantial difference from those commonly understood in the art. The techniques and procedures described or referenced herein are commonly understood and are commonly used by those skilled in the art using conventional methodologies, such as widely used enzyme-linked immunosorbent assays. Where appropriate, procedures involving the use of commercially available kits and reagents are generally performed according to the protocols and / or parameters defined by the manufacturer, unless otherwise stated.

[0053]

[0073] As used herein and in the appended claims, the terms “a,” “an,” and “the” refer to both singular and plural objects unless the context clearly indicates otherwise.

[0054]

[0074] When preceding a number, the term "approximately" means up to ±20% of the number. In some embodiments, "approximately" of a number means up to ±20%, up to ±19%, up to ±18%, up to ±17%, up to ±16%, up to ±15%, up to ±14%, up to ±13%, up to ±12%, up to ±11%, up to ±10%, up to ±9%, up to ±8%, up to ±7%, up to ±6%, up to ±5%, up to ±4%, up to ±3%, up to ±2%, up to ±1%, up to less than ±1%, or any other value or range within that value.

[0055]

[0075] The term "biomarker" refers to an indicator of treatment with a therapeutic agent, such as caveolin-1 peptide or its derivatives, in a subject with a disease or disorder. In some embodiments, biomarkers are used to predict or determine efficacy in a subject with a disease or disorder. In some embodiments, biomarkers can be detected or measured in biological samples obtained from subjects with a disease or disorder. In some embodiments, biomarkers can be detected or measured in biological samples treated in vitro with a therapeutic agent, such as caveolin-1 peptide or its derivatives. In some embodiments, biomarkers can be detected or measured in biological samples obtained from subjects treated in vivo with a therapeutic agent, such as caveolin-1 peptide or its derivatives. Examples of biomarkers include, but are not limited to, DNA, RNA, protein, phosphorylated protein, carbohydrate, or glycolipid-based biomarkers.

[0056]

[0076] As used herein, the letter "p" indicates a protein, specifically a phosphorylated protein, such as phosphorylated SMAD2 / 3 as pSMAD2 / 3. Methods for detecting phosphorylated proteins are well known in the art.

[0057]

[0077] As used herein, the terms “detection” or “detecting” refer to the quantitative or qualitative detection of a biomarker in a biological sample. The terms “detection” or “detecting” include any means of detecting the expression level of a biomarker in a biological sample, such as immunoassay, qRT-PCR, or mass spectrometry.

[0058]

[0078] As used herein, the terms “identify” or “identify” refer to the quantitative or qualitative detection of a biomarker in a biological sample. The terms “identify” or “identify” include any means of identifying the expression level of a biomarker in a biological sample, such as immunoassay, qRT-PCR, or mass spectrometry.

[0059]

[0079] The term “section” of a tissue sample refers to a portion or fragment of a tissue sample, for example, a thin slice of tissue or cells cut from a tissue sample. In some embodiments, the same section of a tissue sample is analyzed at the morphological and molecular levels, or is analyzed for both proteins and nucleic acids for the biomarkers described herein. In some embodiments, multiple sections of tissue are analyzed at the morphological and molecular levels, or is analyzed for both proteins and nucleic acids for the biomarkers described herein.

[0060]

[0080] The term "polynucleotide" or "nucleic acid" refers to a polymer of nucleotide monomers covalently linked together in a chain. Exemplary nucleic acids include DNA and RNA.

[0061]

[0081] The term "amino acid" refers to the structural unit (monomer) that makes up a protein, polypeptide, or peptide. The terms "polypeptide" or "protein" include any polymer of amino acids or amino acid residues. A "peptide" is a small polypeptide with a size of less than approximately 15 to 20 amino acid residues. The term "amino acid sequence" refers to a series of amino acids or amino acid residues.

[0062]

[0082] Methods for determining sequence similarity or identity between two or more nucleic acid sequences or two or more amino acid sequences are known in the art. Sequence similarity or identity can be determined using standard techniques including, but not limited to, the local sequence identity algorithm of Smith & Waterman, Adv.Appl.Math.2, 482 (1981), the sequence identity alignment algorithm of Needleman & Wunsch, J Mol.Biol.48,443 (1970), the similarity search method of Pearson & Lipman, Proc.Natl.Acad.Sci.USA 85,2444 (1988), computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA from Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Drive, Madison, WI), the Best Fit sequencing program described in Devereux et al., Nucl.Acid Res.12, 387-395 (1984), or checks. Another suitable algorithm is the BLAST algorithm described in Altschul et al., J Mol.Biol.215, 403-410, (1990) and Karlin et al., Proc.Natl.Acad.Sci.USA 90, 5873-5787 (1993). An exemplary BLAST program is the WU-BLAST-2 program, obtained from Altschul et al., Methods in Enzymology, 266, 460-480 (1996); blast.wustl / edu / blast / README.html. WU-BLAST-2 uses several search parameters, which are optionally set to default values. The parameters are dynamic values, established by the program itself depending on the composition of a particular sequence and the composition of the particular database from which the target sequence is searched. However, the values ​​may be adjusted to increase sensitivity.Furthermore, an additional algorithm is the gap BLAST reported in Altschul et al, (1997) Nucleic Acids Res.25, 3389-3402. Unless otherwise indicated, the calculation of identity percentages in this disclosure is performed using the BLAST algorithm, which is available at the worldwide web address:blast.ncbi.nlm.nih.gov / Blast.cgi.

[0063]

[0083] The term "antibody" refers to immunoglobulins that may be derived whole or partially from natural sources or produced synthetically. Examples of antibodies include, but are not limited to, monoclonal antibodies, polyclonal antibodies, antibodies with polyepitope specificity, single-chain antibodies, multispecific antibodies, and antibody fragments (e.g., scFv, diabodies, triabodies, minibodies). Such antibodies may be chimeric antibodies, humanized antibodies, human antibodies, and synthetic antibodies.

[0064]

[0084] As used herein, the terms “to treat,” “to treat,” or “treatment,” and their grammatical variations, have the same meaning as generally understood by those skilled in the art. In some embodiments, these terms may refer to an approach to obtain a beneficial or desirable clinical outcome. This may refer to slowing the onset or progression of a condition, disorder, or disease; reducing or alleviating associated symptoms; causing a complete or partial regression of the condition; or any combination thereof. For the purposes of the present invention, beneficial or desirable clinical outcomes include, but are not limited to, symptom reduction or alleviation, a reduction in the severity of the disease, stabilization of the disease state (e.g., no worsening), delay or slowing the progression of the disease, improvement or alleviation of the condition, and remission (partial or complete), whether detectable or undetectable. “To treat,” “to treat,” or “treatment” may also mean extending survival compared to the expected survival time without treatment. Thus, subjects requiring treatment may be subjects already suffering from the disease or disorder in question. The terms “to treat,” “to treat,” or “treatment” include inhibiting or mitigating the increase in the severity of a pathological condition or symptom compared to no treatment, and do not necessarily mean the complete cessation of the associated disease or condition. The terms “to treat,” “to treat,” or “treatment” may also refer to providing a therapeutic activator, such as caveolin-1 peptide or a derivative thereof, to a biological sample obtained from an object having a disease or disorder.

[0065]

[0085] As used herein, the term “pharmaceutically acceptable carrier or excipient” includes, but is not limited to, any auxiliaries, carriers, lubricants, sweeteners, diluents, preservatives, pigments / colorants, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, surfactants, and / or emulsifiers. Exemplary pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth; malt; gelatin; talc; cocoa butter, waxes, animal and vegetable fats, paraffin, silicone, bentonite, silicic acid, zinc oxide; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; water free of pyrogens; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer; and other suitable substances used in pharmaceutical formulations. Unless conventional media and / or agents are incompatible with the agents of this disclosure, their use in therapeutic compositions is intended. Co-active ingredients can also be incorporated into the composition.

[0066]

[0086] The term “interstitial lung disease” or “ILD” refers to a group of lung diseases that affect the interstitium (the tissue and space surrounding the air sacs of the lungs). ILDs may also be classified according to suspected or known causes, or they may be idiopathic. For example, ILDs may be classified as those caused by inhaled substances (inorganic or organic), drug-induced (e.g., antibiotics, chemotherapy drugs, antiarrhythmics, statins), associated with connective tissue diseases (e.g., systemic sclerosis, polymyositis, dermatomyositis, systemic lupus erythematosus, rheumatoid arthritis), associated with lung infections (e.g., atypical pneumonia, Pneumocystis pneumonia, tuberculosis, Chlamydia trachomatis, respiratory syncytial virus, COVID-19), associated with malignancies (e.g., lymphangitis-associated carcinomatosis), or they may be idiopathic (e.g., sarcoidosis, idiopathic pulmonary fibrosis, Hanmann-Ricci syndrome, or antisynthetase syndrome).

[0067]

[0087] The term "idiopathic pulmonary fibrosis" or "IPF" refers to a chronic, progressive form of lung disease characterized by fibrosis of the lung's supporting framework (interstitium). Microscopically, lung tissue from patients with IPF exhibits a set of characteristic histological / pathological features known as typical interstitial pneumonia, characterized by a heterogeneous, patchy appearance with alternating areas of healthy lung tissue, interstitial inflammation, fibrosis, and honeycomb changes. By definition, the term IPF is used when the cause of pulmonary fibrosis is unknown ("idiopathic"). Symptoms typically include gradually developing shortness of breath and a dry cough. Other changes include fatigue, as well as abnormally large, dome-shaped fingernails and toenails (clubbing). Complications may include pulmonary hypertension, heart failure, pneumonia, and pulmonary embolism.

[0068]

[0088] The term "optimal dose" refers to the amount of therapeutic agent that is effective in "alleviating" or "treating" a disease or disorder in a subject. The optimal dose of a therapeutic agent may vary depending on factors such as the individual's disease state, age, sex, and weight. The optimal dose is also the amount at which the therapeutic benefits outweigh any toxic or adverse effects of the therapeutic agent.

[0069]

[0089] The terms "increased expression level" or "enhanced expression level" refer to an increase in the expression of a biomarker (e.g., mRNA or protein biomarker) in a biological sample from a subject treated with a therapeutic agent, compared to a control sample.

[0070]

[0090] The terms "decreased expression level" or "reduced expression level" refer to a decrease in the expression of a biomarker (e.g., mRNA or protein biomarker) in a biological sample from a subject treated with a therapeutic agent, compared to a control sample.

[0071] Therapeutic activators

[0091] This disclosure provides a method for identifying changes in the expression levels of biomarkers associated with caveolin-1 peptide therapy. In some embodiments, the biomarkers described herein are used to determine or predict the efficacy of a therapeutic agent in a subject with a disease or disorder. In some embodiments, the biomarkers described herein are used to determine or predict the optimal dose of a therapeutic agent in a subject with a disease or disorder. In some embodiments, the disease or disorder is an interstitial lung disease, such as idiopathic pulmonary fibrosis. In some embodiments, the therapeutic agent is a caveolin-1 peptide or a derivative thereof.

[0072]

[0092] The term "therapeutic activator" refers to any type of drug, agent, or pharmaceutical used to treat, control, or prevent any disease, disorder, or undesirable condition in a mammalian subject. In some embodiments, the therapeutic activator is a small molecule. In some embodiments, the therapeutic activator is a biological agent. In some embodiments, the therapeutic activator is a caveolin-1 peptide or a derivative thereof. In some embodiments, the therapeutic activator is CSP-7. In some embodiments, the therapeutic activator is Var55.

[0073] Caveolin-1 peptide and its derivatives

[0093] In some embodiments, the therapeutic activator is the caveolin-1 peptide or a derivative thereof. Natural human caveolin-1 has 178 amino acids (see Sequence ID No. 1 in Table 1 below) and a molecular weight of 22 kDa. Caveolin-1 is an intrinsic membrane protein involved in endocytosis, extracellular matrix organization, cholesterol distribution, cell migration, and signaling. See Boscher and Nabi, Adv Exp Med Biol, 2012;729-29-50. Caveolin-1 plays a role in maintaining homeostasis in the fibrotic process by regulating several key signaling components, including TGF-β and ERK1 / 2, and by participating in a series of major regulatory pathways (see Shihata et al., Front.Pharmacol.2017; 8:567; and Gvaramia et al., Matrix Biol, 2013:32(6):307-315). Internalization of TGF-βR associated with non-lipid rafts increases TGF-β signaling, while internalization associated with caveolin-1 increases TGF-βR degradation and therefore reduces or eliminates TGF-β signaling (Zhang et al, JBC, 2005; 280:12239-45; Di Guglielmo et al., Nat Cell Biol, 2003; 5:410-21). Endogenous caveolin-1 has been found to be constitutively suppressed in bleomycin-induced animal models of IPF and in fibrotic lungs of IPF patients (see Wang et al., J Exp Med, 2006; 203(13):2895-2906; Sanders et al., PLoS One, 2015; 10(2), e0116995; and Sanders et al., Am J Respir Cell Mol Biol, 2017; 56(1):50-61). Derivatives of the caveolin-1 peptide include caveolin-1 scaffold domain peptides (CSPs).

[0074]

[0094] In some embodiments, the therapeutic activator is a caveolin-1 scaffold domain peptide (CSP) or a derivative thereof. The caveolin-1 scaffold domain (CSD) consists of amino acids 82-101 of caveolin-1 (see SEQ ID NO: 2 in Table 1 below). Caveolin-1 CSDs play important roles in the dimerization of caveolin-1 and the regulation of various signaling intermediates, many of which are involved in the pathogenesis of pulmonary and histofibrosis (Shetty et al., Am J Respir Cell Mol Biol 2012; 47:474-83; Fridolfsson et al., FASEB J 2014; 28:3823-31; Degryse et al., Am J Physiol Cell Mol Physiol 2010; 299:L442-L452; and Egger et al., PLos One, 2013; 8:e63432). The CSD domain of caveolin-1 has demonstrated inhibition of various factors involved in Wnt signaling, β-catenin-mediated transcription, activation of SRC, EGFR, MEK1, and ERK-2, as well as other factors related to pro-fibrillation signaling (Shetty et al., Am J Respir Cell Mol Biol 2012; 47:474-83; Bhandary et al., Am J Physiol Cell Mol Physiol 2012; 302:L463-L473; Bhandary et al., Am J Pathol 2013; 183:131-143; Fridolfsson et al., FASEB J 2014; 28:3823-31; Degryse et al., Am J Physiol Cell Mol Physiol 2010; 299:L442-L452; and Fiddler et al., Ann Am Thorac (See Soc, 2016; 13:1430-2). The endogenous CSD domain can homodimerize with other caveolin-1 proteins and interact with proteins containing the caveolin-binding domain (CBD) motif.It is estimated that up to 30% of all endogenous proteins possess a CBD motif, and the caveolin-1 CSD domain is hypothesized to provide stability to these proteins (see Marudamuthu et al., Am J Pathol 2015; 185:55-68). Previous studies have shown that CSPs prevent, limit, or reverse pulmonary fibrosis in animal models (see Marudamuthu et al., Sci Transl Med, 2019; 11(522), eaat2848). Treatment with CSP20mer (full-length CSD of caveolin-1) reduced pulmonary αSMA and pulmonary epithelial apoptosis, decreased collagen deposition, and downregulated the expression of pro-fibrosis signaling molecules (see Bhandary et al., Am J Phys Lung Cell Mol Phys, 2012, 302(5), L463-L473; Razani et al., JBC, 2001, 276(9),6727-6738; and Lee et al., Biochem Biophys Res Commun, 2007, 359(2):385-390).

[0075]

[0095] In some embodiments, the therapeutic activator is CSP-7, a 7-amino acid fragment of human CSD of caveolin-1. CSP-7 has been shown to be effective in reducing fibrosis in both in vitro and in vivo models (see Marudamuthu et al., Sci Transl Med, 2019; 11(522), eaat2848). CSP-7 increases p53 protein levels in cells such as fibrotic lung fibroblasts, decreases urokinase plasminogen activator (uPA) and uPA receptor (uPAR), and increases the expression of plasminogen activator inhibitor 1 (PAI-1). See International Publication No. 2014 / 145389A1 and International Publication No. 2020 / 055812A1 (these are incorporated herein by reference in their entirety).

[0076]

[0096] In some embodiments, the therapeutic agent is Var55.

[0077]

[0097] Table 1 below shows exemplary amino acid sequences of caveolin-1 peptides or their derivatives. Uppercase letters indicate L-amino acids, and lowercase letters indicate D-amino acids. The term "Ac" refers to an acetyl group, and the term "NH2" refers to an amino group. "O" represents ornithine.

[0078] [Table 1]

[0079] [Table 2]

[0080] [Table 3]

[0081]

[0098] In some embodiments, the Cav-1 peptide or modified Cav-1 peptide is (a) Consists of any one of the amino acid sequences of SEQ ID NOs: 1 to 110; (b) containing one core sequence of any of the amino acid sequences of SEQ ID NOs: 1 to 110; or (c) comprising one core sequence of any of the amino acid sequences of SEQ ID NOs: 1 to 110, wherein the core sequence includes one or more amino acid substitutions, insertions, deletions, or chemical modifications.

[0082]

[0099] In some embodiments, the Cav-1 peptide or modified Cav-1 peptide includes one core sequence of any of the amino acid sequences of SEQ ID NOs: 1 to 110. In some embodiments, the Cav-1 peptide or modified Cav-1 peptide includes one core sequence of any of the amino acid sequences of SEQ ID NOs: 1 to 110, wherein the core sequence includes one or more amino acid substitutions, insertions, deletions, or chemical modifications. In some embodiments, the core sequence is SEQ ID NO: 3. In some embodiments, the core sequence is SEQ ID NO: 6.

[0083]

[0100] In some embodiments, the therapeutic activator is a caveolin-1 peptide or a derivative thereof. In some embodiments, the caveolin-1 peptide or a derivative thereof contains or consists of one of the amino acid sequences of SEQ ID NOs: 1 to 110. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with one of SEQ ID NOs: 1 to 110. In some embodiments, the caveolin-1 peptide or a derivative thereof contains one of the amino acid sequences of SEQ ID NOs: 1 to 110 having one or more mutations thereto. For example, in some embodiments, the caveolin-1 peptide or a derivative thereof contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations with respect to one of SEQ ID NOs: 1 to 110. In some embodiments, the caveolin-1 peptide or its derivative comprises one of the amino acid sequences of SEQ ID NOs: 1 to 110 having 1 to 5, 5 to 10, or 11 to 15, or more mutations. In some embodiments, the caveolin-1 peptide or its derivative comprises 1 to 5 additional amino acids at either the N-terminus or C-terminus, or both, of one of the SEQ ID NOs: 1 to 110.

[0084]

[0101] In some embodiments, the therapeutic activator is a caveolin-1 peptide or a derivative thereof. In some embodiments, the caveolin-1 peptide or a derivative thereof contains or comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with respect to SEQ ID NO: 1. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence of SEQ ID NO: 1 having one or more mutations thereto. For example, in some embodiments, the caveolin-1 peptide or a derivative thereof contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations with respect to SEQ ID NO: 1. In some embodiments, the caveolin-1 peptide or its derivative comprises the amino acid sequence of SEQ ID NO: 1 having 1 to 5, 5 to 10, 11 to 5, 15 to 20, 10 to 25, 25 to 30, or more than 30 mutations.

[0085]

[0102] In some embodiments, the therapeutic activator is a caveolin-1 peptide or a derivative thereof. In some embodiments, the caveolin-1 peptide or a derivative thereof contains or comprises the amino acid sequence of SEQ ID NO: 2. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% identity with respect to SEQ ID NO: 2. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence of SEQ ID NO: 2 having one or more mutations thereto. For example, in some embodiments, the caveolin-1 peptide or a derivative thereof contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations with respect to SEQ ID NO: 2. In some embodiments, the caveolin-1 peptide or its derivative comprises the amino acid sequence of SEQ ID NO: 2 having 1 to 5, 5 to 10, 11 to 15, or more mutations. In some embodiments, the caveolin-1 peptide or its derivative comprises 1 to 5 additional amino acids at either the N-terminus or C-terminus, or both, of SEQ ID NO: 2.

[0086]

[0103] In some embodiments, the therapeutic activator is a caveolin-1 peptide or a derivative thereof. In some embodiments, the caveolin-1 peptide or a derivative thereof contains or comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 85% identity with respect to SEQ ID NO: 3. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence of SEQ ID NO: 3 having one or more mutations thereto. For example, in some embodiments, the caveolin-1 peptide or a derivative thereof contains 1, 2, 3, 4, or 5 mutations with respect to SEQ ID NO: 3. In some embodiments, the caveolin-1 peptide or a derivative thereof contains 1 to 5 additional amino acids at either the N-terminus or C-terminus, or both, of SEQ ID NO: 3.

[0087]

[0104] In some embodiments, the therapeutic activator is a caveolin-1 peptide or a derivative thereof. In some embodiments, the caveolin-1 peptide or a derivative thereof contains or comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence having at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95% identity with respect to SEQ ID NO: 4. In some embodiments, the caveolin-1 peptide or a derivative thereof contains an amino acid sequence of SEQ ID NO: 4 having one or more mutations thereto. For example, in some embodiments, the caveolin-1 peptide or a derivative thereof contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations with respect to SEQ ID NO: 4. In some embodiments, the caveolin-1 peptide or its derivative comprises the amino acid sequence of SEQ ID NO: 4 having 1 to 5, 5 to 10, 11 to 15, or more mutations. In some embodiments, the caveolin-1 peptide or its derivative comprises 1 to 5 additional amino acids at either the N-terminus or C-terminus, or both, of SEQ ID NO: 4.

[0088]

[0105] In some embodiments, the caveolin-1 peptide or its derivative comprises at least one non-standard amino acid. In some embodiments, the caveolin-1 peptide or its derivative comprises two or more non-standard amino acids. In some embodiments, the caveolin-1 peptide or its derivative comprises four or more non-standard amino acids. In some embodiments, the non-standard amino acid is ornithine. In some embodiments, the non-standard amino acid is D-alanine. In some embodiments, the caveolin-1 peptide or its derivative comprises one or more non-standard amino acids, where the non-standard amino acid is ornithine.

[0089]

[0106] In some embodiments, the caveolin-1 peptide or its derivative comprises an L-amino acid. In some embodiments, the caveolin-1 peptide or its derivative comprises a D-amino acid. In some embodiments, the caveolin-1 peptide or its derivative comprises an L-amino acid and a D-amino acid. In some embodiments, the caveolin-1 peptide or its derivative comprises one or more D-amino acids, where the D-amino acid is D-alanine.

[0090]

[0107] In some embodiments, the caveolin-1 peptide or its derivative includes N- or C-terminal modifications. In some embodiments, the caveolin-1 peptide or its derivative includes N-terminal modifications. In some embodiments, the caveolin-1 peptide or its derivative includes C-terminal modifications. In some embodiments, the N-terminal modification is acylation. In some embodiments, the C-terminal modification is amidation.

[0091]

[0108] In some embodiments, the caveolin-1 peptide or its derivatives include cell-permeable peptides (CPPs). Examples of CPPs are shown in Table 3 below.

[0092] [Table 4]

[0093]

[0109] In some embodiments, the caveolin-1 peptide or its derivatives are conjugated to a heterogeneous polypeptide segment or polymer, such as polyethylene glycol (PEG). The caveolin-1 peptide or its derivatives may be linked to PEG to increase serum persistence.

[0094]

[0110] In some embodiments, the therapeutic agent is a peptide mimetic compound that mimics the biological effects of a natural caveolin-1 peptide or its derivative. The peptide mimetic may be a non-natural peptide or non-peptide agent that replicates the steric spatial properties of the binding element of a natural caveolin-1 peptide or its derivative so as to have the binding activity and biological activity of a natural caveolin-1 peptide or its derivative.

[0095] Other types of therapeutic agents

[0111] In some embodiments, the therapeutic activator is a drug that reduces or inhibits the activity of transforming growth factor-β (TGF-β). Examples of therapeutic activators that reduce or inhibit TGF-β activity include, but are not limited to, GC-1008 (fresolimmab, Genzyme / MedImmune), reldelimumab (CAT-152; Trabio, Cambridge Antibody), meterimumab (CAT-192, Trabio, Cambridge Antibody), LY-2157299 (Eli Lilly), and ACU-HTR-028 (Opko Health), as well as antibodies targeting one or more TGF-β isoforms, and inhibitors of TGF-β receptor kinase TGFBR1 (ALK5) and TGFBR2.

[0096]

[0112] In some embodiments, the therapeutic activator is pirfenidone (also known as Esbriet). Pirfenidone is a small molecule with anti-fibrotic and anti-inflammatory properties. Pirfenidone has been shown to downregulate growth factors (e.g., TGF-β), procollagen I and II, and inflammatory mediators (e.g., TNF-α and IL-β).

[0097]

[0113] In some embodiments, the therapeutic agent is an endothelin receptor antagonist. In some embodiments, the endothelin receptor antagonist targets both endothelin receptor A and endothelin receptor B. In some embodiments, the endothelin receptor antagonist targets endothelin receptor A. Examples of endothelin receptor antagonists include, but are not limited to, ambrisentan, avosentan, bosentan, clazosentan, dalsenta, BQ-153, FR-139317, L-744453, macitentan, PD-145065, PD-156252, PD163610, PS-433540, S-0139, sitaxentan sodium, TBC-3711, and dibotentan.

[0098]

[0114] In some embodiments, the therapeutic activator is a drug that reduces or inhibits the activity of connective tissue growth factor (CTGF). Examples of therapeutic activators that reduce or inhibit CTGF activity include, but are not limited to, FG-3019 and FibroGen, CTGF neutralizing antibodies, and matrix metalloproteinase (MMP) inhibitors such as MMP-12, PUP-1, and tigapotide triflutate.

[0099]

[0115] In some embodiments, the therapeutic activator is a drug that reduces or inhibits the activity of the epidermal growth factor receptor (EGFR). Examples of therapeutic activators that reduce or inhibit EGFR activity include, but are not limited to, erlotinib, gefitinib, BMS-690514, cetuximab, antibodies targeting EGFR, inhibitors of EGFR kinases, and modulators of post-receptor signaling pathways.

[0100]

[0116] In some embodiments, the therapeutic activator is a drug that reduces or inhibits the activity of platelet-derived growth factor (PDGF). Examples of therapeutic activators that reduce or inhibit PDGF activity include, but are not limited to, imatinib mesylate (Novartis), PDGF neutralizing antibodies, antibodies that target the PDGF receptor (PDGFR), inhibitors of PDGFR kinase activity, and post-receptor signaling pathways.

[0101]

[0117] In some embodiments, the therapeutic activator is a drug that reduces or inhibits the activity of vascular endothelial growth factor (VEGF). Examples of therapeutic activators that reduce or inhibit VEGF activity include, but are not limited to, axitinib, bevacizumab, BIBF-1120, CDP-791, CT-322, IMC-18F1, PTC-299, ramucirumab, VEGF neutralizing antibodies, antibodies targeting VEGF receptor 1 (VEGFR1, Flt-1) and VEGF receptor 2 (VEGFR2, KDR), soluble VEGFR1 (sFlt) and its derivatives that neutralize VEGF, and inhibitors of VEGF receptor kinase activity.

[0102]

[0118] In some embodiments, the therapeutic activator is a drug that inhibits multiple receptor kinases. For example, BIBF-1120 (also known herein as nintedanib) is a therapeutic activator that inhibits receptor kinases for vascular endothelial growth factor, fibroblast growth factor, and platelet-derived growth factor.

[0103]

[0119] In some embodiments, the therapeutic activator is a drug that interferes with integrin function. Examples of therapeutic activators that interfere with integrin function include, but are not limited to, STX-100, IMGN-388, and integrin-targeting antibodies.

[0104]

[0120] In some embodiments, the therapeutic activator is a drug that interferes with the pro-fibrotic activity of IL-4 and IL-13. Examples of therapeutic activators that interfere with the pro-fibrotic activity of IL-4 and IL-13 include, but are not limited to, AER-001, AMG-317, APG-201, sIL-4Rα, AER-001, AMG-317, anlukinzumab, CAT-354, syntredekinbesdotox, MK-6105, QAX-576, SB-313, SL-102, TNX-650, tralokinumab, lebrikizumab, SAR156597, antibodies targeting the IL-4 receptor or IL-13 receptor, soluble IL-4 receptors or derivatives thereof that bind to and neutralize both IL-4 and IL-13, chimeric proteins containing all or part of IL-13, and inhibitors of the JAK-STAT kinase pathway.

[0105]

[0121] In some embodiments, the therapeutic activator is a CCL-2 inhibitor, such as an anti-CCL2 antibody (e.g., CNT0888).

[0106]

[0122] In some embodiments, the therapeutic activator is a drug that interferes with epithelial-mesenchymal transition, such as an mTOR inhibitor (including, but not limited to, AP-23573 and rapamycin).

[0107]

[0123] In some embodiments, the therapeutic activator is a drug that reduces copper levels, such as tetrathiomolybdate.

[0108]

[0124] In some embodiments, the therapeutic activator is a drug that reduces oxidative stress. Examples of therapeutic activators that reduce oxidative stress include, but are not limited to, N-acetylcysteine, tetrathiomolybdate, and interferon gamma.

[0109]

[0125] In some embodiments, the therapeutic activator is a phosphodiesterase 4 inhibitor (e.g., roflumilast); a phosphodiesterase 5 inhibitor (e.g., myrodenafil, PF-4480682, sildenafil citrate, SLx-2101, tadalafil, udenafil, UK-369003, vardenafil, and zaprinast); or an arachidonic acid pathway modifier, including cyclooxygenase and 5-lipoxegagenase inhibitors (e.g., diloton).

[0110]

[0126] In some embodiments, therapeutic activators reduce tissue remodeling or fibrosis. Examples of therapeutic activators that reduce tissue remodeling or fibrosis include, but are not limited to, prolyl hydrolase inhibitors (e.g., 1016548, CG-0089, FG-2216, FG-4497, FG-5615, FG-6513, fibrostatin A (Takeda), rufironil, P-1894B, and safironil), peroxisome proliferator-activated receptor (PPAR) gamma agonists (e.g., pioglitazone and rosiglitazone), carbon monoxide, and doxycycline.

[0111]

[0127] In some embodiments, the therapeutic agent is a drug having antioxidant activity, immunosuppressive activity, and / or anti-inflammatory activity, such as N-acetylcysteine.

[0112]

[0128] In some embodiments, the therapeutic activator is a somatostatin receptor inhibitor, such as a somatostatin analog (e.g., SOM230 and octreotide).

[0113]

[0129] In some embodiments, the therapeutic agent is a drug having anti-angiogenic activity, immunomodulatory activity, and / or anti-inflammatory activity, such as thalidomide or minocycline.

[0114]

[0130] In some embodiments, the therapeutic activator is an agent that inhibits the enzyme lysyl oxidase-like 2 (LOXL2), such as an anti-LOXL2 antibody (e.g., GS-6624).

[0115]

[0131] In some embodiments, the therapeutic activator is a drug that targets the renin-angiotensin system (e.g., losartan).

[0116]

[0132] Other examples of therapeutic activators for use in this disclosure are described, for example, in Rafii et al., J. Thorac Dis, 2013; 5(1):48-73; U.S. Patent Application Publication No. 2019 / 0062836A1; U.S. Patent Application Publication No. 2012 / 0014917A1; International Publication No. 2014 / 145389A1 and International Publication No. 2020 / 055812A1, which are incorporated herein by reference in their entirety.

[0117] Treatment methods using therapeutic activators

[0133] This disclosure provides biomarkers related to therapeutic agents in subjects with a disease or disorder (e.g., interstitial lung disease). In some embodiments, one or more therapeutic agents may be administered by any of the methods disclosed herein.

[0118]

[0134] In some embodiments, the therapeutic activator described herein is administered in vitro to a biological sample. In some embodiments, the therapeutic activator is administered in vitro to a biological sample obtained from a subject having a disease or disorder. In some embodiments, the therapeutic activator is administered in vitro to a biological sample obtained from a subject having interstitial lung disease. In some embodiments, the therapeutic activator is administered in vitro to a biological sample obtained from a subject having idiopathic pulmonary fibrosis. In some embodiments, the biological sample is fibroblasts isolated from a subject having idiopathic pulmonary fibrosis. In some embodiments, the biological sample is lung tissue isolated from a subject having idiopathic pulmonary fibrosis. In some embodiments, the biological sample is basal-like cells isolated from a subject having idiopathic pulmonary fibrosis. In some embodiments, the therapeutic activator is caveolin-1 peptide or a derivative thereof (e.g., CSP-7).

[0119]

[0135] In some embodiments, the therapeutic activator is administered in vivo to subjects with a disease or disorder. In some embodiments, the therapeutic activator is administered in vivo to subjects with interstitial lung disease. In some embodiments, the therapeutic activator is administered in vivo to subjects with idiopathic pulmonary fibrosis. In some embodiments, caveolin-1 peptide or its derivative (e.g., CSP-7) is administered in vivo to subjects with idiopathic pulmonary fibrosis.

[0120]

[0136] In some embodiments, the therapeutic activator (e.g., caveolin-1 peptide or a derivative thereof) comprises at least one pharmaceutically acceptable carrier or excipient. In some embodiments, the therapeutic activator comprising at least one pharmaceutically acceptable carrier or excipient is administered to a biological sample in vitro. In some embodiments, the therapeutic activator comprising at least one pharmaceutically acceptable carrier or excipient is administered in vivo to a subject with a disease or disorder.

[0121]

[0137] In some embodiments, the therapeutic activator (e.g., caveolin-1 peptide or its derivatives) is administered in the absence of a pharmaceutically acceptable carrier or excipient. In some embodiments, the therapeutic activator is administered in vitro to a biological sample in the absence of a pharmaceutically acceptable carrier or excipient. In some embodiments, the therapeutic activator is administered in vivo to a subject with a disease or disorder in the absence of a pharmaceutically acceptable carrier or excipient.

[0122]

[0138] In some embodiments, the therapeutic activator is administered to subjects with a disease or disorder, such as interstitial lung disease, orally, parenterally, sublingually, percutaneously, rectally, transmucosally, topically, by inhalation, buccally, intrathoracically, intravenously, intraarterially, intragastricly, intraperitoneally, subcutaneously, intramuscularly, intranasally, subarachnoidally, and intraarticularly, or in combination thereof.

[0123]

[0139] In some embodiments, caveolin-1 peptide or its derivatives are administered to subjects with idiopathic pulmonary fibrosis orally, parenterally, sublingually, percutaneously, rectally, transmucosally, topically, by inhalation, buccally, intrathoracically, intravenously, intraarterially, intragastricly, intraperitoneally, subcutaneously, intramuscularly, intranasally, subarachnoidally, and intraarticularly, or in combination thereof. In some embodiments, caveolin-1 peptide or its derivatives are administered intravenously to subjects with idiopathic pulmonary fibrosis. In some embodiments, caveolin-1 peptide or its derivatives are administered by inhalation to subjects with idiopathic pulmonary fibrosis. In some embodiments, caveolin-1 peptide or its derivatives are administered intranasally to subjects with idiopathic pulmonary fibrosis. In some embodiments, caveolin-1 peptide or its derivatives are CSP-7. In some embodiments, caveolin-1 peptide or its derivatives are Var55.

[0124]

[0140] In some embodiments, the therapeutic activator is administered to subjects with the disease or disorder once daily, twice daily, three times daily, once weekly, twice weekly, three times weekly, five times weekly, once every two weeks, once every three weeks, once a month, once every two months, once every three months, or once every six months. In some embodiments, the therapeutic activator is administered to subjects with interstitial lung disease once daily, twice daily, three times daily, once weekly, twice weekly, three times weekly, five times weekly, once every two weeks, once every three weeks, once a month, once every two months, once every three months, or once every six months. In some embodiments, the therapeutic activator is administered to subjects with idiopathic pulmonary fibrosis once daily, twice daily, three times daily, once a week, twice a week, three times a week, five times a week, once every two weeks, once every three weeks, once a month, once every two months, once every three months, or once every six months.

[0125]

[0141] In some embodiments, caveolin-1 peptide or its derivatives are administered to subjects with disease or impairment once daily, twice daily, three times daily, once weekly, twice weekly, three times weekly, five times weekly, once every two weeks, once every three weeks, once a month, once every two months, once every three months, or once every six months. In some embodiments, caveolin-1 peptide or its derivatives are administered to subjects with interstitial lung disease once daily, twice daily, three times daily, once weekly, twice weekly, three times weekly, five times weekly, once every two weeks, once every three weeks, once a month, once every two months, once every three months, or once every six months. In some embodiments, the caveolin-1 peptide or its derivative is administered to subjects with idiopathic pulmonary fibrosis once daily, twice daily, three times daily, once weekly, twice weekly, three times weekly, five times weekly, once every two weeks, once every three weeks, once monthly, once every two months, once every three months, or once every six months. In some embodiments, the caveolin-1 peptide or its derivative is CSP-7. In some embodiments, the caveolin-1 peptide or its derivative is Var55.

[0126]

[0142] In some embodiments, the therapeutic activator is administered to subjects with the disease or disorder at a dose of approximately 0.01 mg / kg to approximately 250 mg / kg. In some embodiments, the therapeutic activator is administered to subjects with the disease or disorder at a dose of approximately 0.01 mg / kg to approximately 50 mg / kg. In some embodiments, the therapeutic activator is administered to subjects with the disease or disorder at a dose of approximately 0.05 mg / kg to approximately 50 mg / kg. In some embodiments, the therapeutic agent is administered to a subject with a disease or disorder at approximately 0.01 mg / kg, approximately 0.02 mg / kg, approximately 0.03 mg / kg, approximately 0.04 mg / kg, approximately 0.05 mg / kg, approximately 0.06 mg / kg, approximately 0.07 mg / kg, approximately 0.08 mg / kg, approximately 0.09 mg / kg, approximately 0.1 mg / kg, approximately 0.2 mg / kg, approximately 0.3 mg / kg, approximately 0.4 mg / kg, approximately 0.5 mg / kg, approximately 0.6 mg / kg kg, approximately 0.7mg / kg, approximately 0.8mg / kg, approximately 0.9mg / kg, approximately 1mg / kg, approximately 2mg / kg, approximately 3mg / kg, approximately 4mg / kg, approximately 5mg / kg, approximately 6mg / kg, approximately 7mg / kg, approximately 8mg / k g, about 9mg / kg, about 10mg / kg, about 11mg / kg, about 12mg / kg, about 13mg / kg, about 14mg / kg, about 15mg / kg, about 16mg / kg, about 17mg / kg, about 18mg / kg, about 19m g / kg, about 20mg / kg, about 21mg / kg, about 22mg / kg, about 23mg / kg, about 24mg / kg, about 25mg / kg, about 26mg / kg, about 27mg / kg, about 28mg / kg, about 29mg / kg, Approximately 30mg / kg, approximately 31mg / kg, approximately 32mg / kg, approximately 33mg / kg, approximately 34mg / kg, approximately 35mg / kg, approximately 36mg / kg, approximately 37mg / kg, approximately 38mg / kg, approximately 39mg / kg, approximately 40mg It is administered in doses of approximately 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg, 75 mg / kg, 100 mg / kg, 125 mg / kg, 150 mg / kg, 175 mg / kg, 200 mg / kg, 225 mg / kg, 250 mg / kg, or higher.

[0127]

[0143] In some embodiments, caveolin-1 peptide or its derivatives is administered to subjects with idiopathic pulmonary fibrosis at doses of approximately 0.01 mg / kg, approximately 0.05 mg / kg, approximately 0.06 mg / kg, approximately 0.07 mg / kg, approximately 0.08 mg / kg, approximately 0.09 mg / kg, approximately 0.1 mg / kg, approximately 0.2 mg / kg, approximately 0.3 mg / kg, approximately 0.4 mg / kg, approximately 0.5 mg / kg, approximately 0.6 mg / kg, approximately 0.7 mg / kg, and approximately 0. .8mg / kg, about 0.9mg / kg, about 1mg / kg, about 2mg / kg, about 3mg / kg, about 4mg / kg, about 5mg / kg, about 6mg / kg, about 7mg / kg, about 8mg / kg, about 9mg / kg, about 1 0mg / kg, about 11mg / kg, about 12mg / kg, about 13mg / kg, about 14mg / kg, about 15mg / kg, about 16mg / kg, about 17mg / kg, about 18mg / kg, about 19mg / kg, about 20mg / kg, about 21mg / kg, about 22mg / kg, about 23mg / kg, about 24mg / kg, about 25mg / kg, about 26mg / kg, about 27mg / kg, about 28mg / kg, about 29mg / kg, about 30mg / k g, about 31mg / kg, about 32mg / kg, about 33mg / kg, about 34mg / kg, about 35mg / kg, about 36mg / kg, about 37mg / kg, about 38mg / kg, about 39mg / kg, about 40mg / kg, about It is administered in doses of 41 mg / kg, approximately 42 mg / kg, approximately 43 mg / kg, approximately 44 mg / kg, approximately 45 mg / kg, approximately 46 mg / kg, approximately 47 mg / kg, approximately 48 mg / kg, approximately 49 mg / kg, approximately 50 mg / kg, approximately 75 mg / kg, approximately 100 mg / kg, approximately 125 mg / kg, approximately 150 mg / kg, approximately 175 mg / kg, approximately 200 mg / kg, approximately 225 mg / kg, approximately 250 mg / kg, or higher. In some embodiments, the caveolin-1 peptide or its derivative is CSP-7. In some embodiments, the caveolin-1 peptide or its derivative is Var55.

[0128] Biomarkers for Caveolin-1 Peptide Therapy

[0144] This disclosure provides biomarkers associated with therapeutic agents in subjects with disease or disorder. In some embodiments, the biomarkers are associated with caveolin-1 peptide or derivatives thereof in subjects with interstitial lung disease, such as idiopathic pulmonary fibrosis.

[0129]

[0145] Table 4 below shows exemplary biomarkers for caveolin-1 therapy in patients with interstitial lung disease.

[0130] [Table 5]

[0131] [Table 6]

[0132]

[0146] In some embodiments, the biomarker is selected from the group consisting of MYDGF, soluble RAGE, phosphorylated SMAD2 (pSMAD2), pSMAD3, PDGFRβ, phosphorylated PDGFRβ (pPDGFRβ), PROF1, CALM2, CALR, PPIA, EIF5A, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, soluble CD163 (sCD163), or phosphorylated mTOR (p-mTOR). In some embodiments, the biomarker is selected from the group consisting of MYDGF, soluble RAGE, phosphorylated SMAD2 (pSMAD2), pSMAD3, PDGFβ, PROF1, CALM2, CALR, PPIA, EIF5A, LGALS1, ANXA5, FABP5, GSTP1, ENO1, and YWHAG. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 and / or pSMAD3. In some embodiments, the biomarker is PDGFRβ or pPDGFRβ. In some embodiments, the biomarker is PROF1. In some embodiments, the biomarker is CALM2. In some embodiments, the biomarker is CALR. In some embodiments, the biomarker is PPIA. In some embodiments, the biomarker is EIF5A. In some embodiments, the biomarker is LGALS1. In some embodiments, the biomarker is ANXA5. In some embodiments, the biomarker is FABP5. In some embodiments, the biomarker is GSTP1. In some embodiments, the biomarker is ENO1. In some embodiments, the biomarker is YWHAG. In some embodiments, the biomarker is LGALS7. In some embodiments, the biomarker is p-mTOR. In some embodiments, the biomarker is IL-11. In some embodiments, the biomarker is MMP-2. In some embodiments, the biomarker is CXCL7.In some embodiments, the biomarker is CD163.

[0133]

[0147] MYDGF is a paracrine protein primarily secreted by monocytes and macrophages. MYDGF is also expressed by eosinophils, myeloid cells, synovial cells, cholangiocarcinoma cells, and hepatocellular carcinoma cells. MYDGF promotes tissue repair and cardiac function after myocardial infarction, thereby protecting against cardiac injury. MYDGF-mediated cytoprotection is associated with AKT phosphorylation and inhibition of apoptosis, and MYDGF has also been shown to promote the proliferation of human coronary artery endothelial cells. MYDGF-deficient mice exhibited larger infarct scars and reduced cell proliferation and angiogenesis in the infarct border region compared to wild-type mice. See Korf-Klingebiel et al., Nat.Med.21:140-149 (2015).

[0134]

[0148] Advanced glycation end product-specific receptors (RAGEs) are a member of the immunoglobulin superfamily of cell surface receptors. They are multiligand receptors that bind to AGEs and other molecules, regulating inflammation, oxidative stress, homeostasis, and development. RAGEs are abundantly expressed in normal lung cells, particularly type 1 lung cells. Previous studies have shown that RAGE expression is reduced in lung tissue in IPF compared to controls, and that functional polymorphisms in the RAGE gene are associated with the risk of IPF. See Yamaguchi et al., Respirology, 2017;22:965-71; Ishikawa et al., Respir Res, 2010;11:123; Manichaikul et al., Ann Am Thorac Soc, 2017;14:628-35; Konishi et al., Am J Respir Crit Care Med, 2009; 180:167-75; and Yamaguchi et al., Respir Res, 2020, 21:145.

[0135]

[0149] Soluble RAGE lacks a transmembrane domain but possesses an extracellular ligand-binding domain. Several types of soluble RAGE exist, including endogenous secreted RAGE (esRAGE), a splice variant of RAGE, and deciduous RAGE derived from cell surface RAGE. Soluble RAGE possesses anti-inflammatory properties and functions as a decoy receptor by neutralizing RAGE ligands. Previous studies, using proteomic analysis, have shown that soluble RAGE expression is reduced in lung tissue of subjects with IPF but not in chronic obstructive pulmonary disease (COPD), suggesting a possible link between soluble RAGE and the pathophysiology of IPF (see Ishikawa et al., Respir Res, 2010, 11:123). Other studies have shown reduced soluble RAGE in lung tissue, serum, and BAL of subjects with IPF (see Yamaguchi et al., Respiratory Research, 2020; 21:145).

[0136]

[0150] Mothers-against decapentaplesic homolog (SMAD) proteins are signaling molecules and transcriptional regulators that mediate multiple signaling pathways. Both SMAD2 and SMAD3 mediate TGF-β signaling, thereby regulating several important cellular processes, including cell proliferation, apoptosis, and differentiation. SMAD2 and SMAD3 are recruited to the TGF-β receptor through interaction with the SMAD anchor (SARA) protein for receptor activation and are phosphorylated by the TGF-β receptor. Phosphorylation induces the dissociation of SMAD2 / 3 and SARA, and binding to family member SMAD4. The association with SMAD4 is crucial for the translocation of this protein to the nucleus, where it regulates numerous target genes involved in matrix expression, proliferation, and cell differentiation. See Gauldie et al., Proc Am Thorac Soc, 2006; 3(8):696-702.

[0137]

[0151] Platelet-derived growth factor B (PDGFB) belongs to a family of proteins composed of other PDGFs and vascular endothelial growth factor (VEGF). The PDGFB preproprotein is proteolytically degraded to produce platelet-derived growth factor subunit B, which can homodimerize or heterodimerize with the associated platelet-derived growth factor subunit A. These proteins bind to and activate PDGF receptor tyrosine kinase, which plays a role in a wide range of developmental processes. PDGF signals via platelet-derived growth factor receptor beta (PDGFRB). PDGF signaling has been associated with the pathogenesis of pulmonary fibrosis. Many fibrillation mediators, such as TGF-β, IL-1, TNF-α, bFGF, and thrombin, exhibit PDGF-dependent pro-fibrotic activity. Inhibition of PDGFRB improved bleomycin-induced pulmonary fibrosis in mice. See Abdollahi et al., JEM, 2005 201(6):925-935; and Kishi et al., PLoS One, 2018, 13(12):e0209786.

[0138]

[0152] Profilin-1 (PROF1) is a member of the prolifin family of small actin-binding proteins. Profilin-1 plays a crucial role in actin dynamics by regulating actin polymerization in response to extracellular signals. Prolifin-1 also influences the structure of the cytoskeleton. Profilin-1 deletion is associated with Miller-Dieker syndrome and may also play a role in Huntington's disease.

[0139]

[0153] Calmodulin (CALM2) mediates the regulation of numerous enzymes, ion channels, aquaporins, and other proteins via calcium binding. Many protein kinases and phosphatases are also stimulated by the calmodulin-calcium complex. Calmodulin, along with CCP110 and centrin, is involved in genetic pathways that regulate centrosome cycles and progression through cytokinesis (see Tsang et al., Mol Biol Cell, 2006, 17:3423-3434).

[0140]

[0154] Calreticulin (CALR) is a calcium-binding chaperone that facilitates folding, oligomer assembly, and quality control in the endoplasmic reticulum (ER) via the calreticulin / calnexin cycle. This lectin transiently interacts with almost all monoglycosylated glycoproteins synthesized in the ER. Calreticulin also interacts with the DNA-binding domain of NR3C1 to mediate its nuclear export.

[0141]

[0155] Peptidyl-prolyl cis-trans isomerase A (PPIA) belongs to the peptidyl-prolyl cis-trans isomerase (PPIase) family. PPIases catalyze the cis-trans isomerization of the prolineimide peptide bond of oligopeptides, accelerating protein folding. PPIA is also a cyclosporine-binding protein and may play a role in cyclosporine A-mediated immunosuppression.

[0142]

[0156] Eukaryotic translation initiation factor 5A (EIF5A) is an mRNA-binding protein involved in translation elongation. EIF5A also plays a crucial role in mRNA turnover. EIF5A is involved in actin dynamics and cell cycle progression, and regulates p53 / TP53-dependent apoptosis and TNFα-mediated apoptosis.

[0143]

[0157] Galectin-1 (LGALS1) binds to β-galactosides and various complex carbohydrates. Galectin-1 plays a role in regulating apoptosis, cell proliferation, and cell differentiation. Galectin-1 has previously been shown to inhibit CD45 phosphatase activity and Lyn kinase dephosphorylation. Galectin-1 is a potent inducer of T cell apoptosis.

[0144]

[0158] Annexin A5 (ANXA5) belongs to the annexin family of calcium-dependent phospholipid-binding proteins. Annexin A5 is a calcium channel-active phospholipase A2 and protein kinase C inhibitory protein, and is associated with cell signaling, inflammation, growth, and differentiation.

[0145]

[0159] Fatty acid-binding protein 5 (FABP5) belongs to a small, highly conserved family of cytoplasmic proteins that bind to long-chain fatty acids and other hydrophobic ligands. FABPs can play a role in fatty acid uptake, transport, and metabolism. In addition to cytosolic transport, FABP5 selectively delivers specific fatty acids from the cytosol to the nucleus, where they activate nuclear receptors. FABP5 also delivers retinoic acid to the nuclear receptor peroxisome proliferator-activated receptor delta, which promotes proliferation and survival.

[0146]

[0160] Glutathione S-transferase P (GSTP1) is a member of the GST superfamily, which is widely expressed in mammals. GSTP has the function of deprotonating glutathione, enabling the formation of thioether bonds with electrophilic substrates. In addition to catalytic detoxification, other properties of GSTP1 include chaperone function, regulation of the nitric oxide pathway, regulation of various kinase signaling pathways, and involvement in the forward reaction of protein S-glutathonylation. See Zhang et al., Advances in Cancer Res, 2014, 122:143-175.

[0147]

[0161] Alpha-enolase 1 (ENO1) is a glycolytic enzyme that catalyzes the conversion of 2-phosphoglycerate to phosphoenolpyruvate. In addition to glycolysis, ENO1 is involved in a variety of cellular processes, including plasminogen binding, maintenance of mitochondrial membrane stability, RNA chaperone activity, and signal transduction (see Didiasova et al., Front Cell Dev Biol, 2019).

[0148]

[0162] 14-3-3 protein gamma (YWHAG) is an adapter protein involved in the broad spectrum of regulation of both general and specialized signaling pathways. 14-3-3 protein gamma typically binds to numerous protein partners through recognition of phosphoserine or phosphothreonine motifs, and this binding generally results in the regulation of the activity of the binding partner.

[0149]

[0163] Galectin-7 (LGALS7) belongs to a family of β-galactoside-binding proteins involved in regulating intercellular and cell-matrix interactions for normal cell proliferation control. Galectin-7 has many functions, including roles in cell adhesion, migration, and immune cell regulation.

[0150]

[0164] mTOR is a serine / threonine protein kinase and a central regulator of cellular metabolism, growth, proliferation, and survival. mTOR is a catalytic subunit of two distinct protein complexes, mTORC1 and mTORC2, and is involved in the regulation of at least 800 different proteins. Phosphorylation of mTOR enhances its activity, thereby promoting the activation of downstream signaling pathways.

[0151]

[0165] Interleukin-11 (IL-11) is a pleiotropic cytokine belonging to the IL-6 family of cytokines. Non-standard IL-11 signaling causes pathological conditions common to all fibroinflammatory diseases, including myofibroblast activation, parenchymal cell dysfunction, and inflammation, and also inhibits tissue regeneration (see Cook and Schafer, Annual Review of Medicine, 2020; 71:263-276).

[0152]

[0166] MMP-2 belongs to the matrix metalloproteinase family, which are zinc-dependent enzymes capable of cleaving components and signaling molecules in the extracellular matrix. MMP-2 is considered the most ubiquitous metalloproteinase in this enzyme family and is involved in many diverse cellular functions, including the regulation of angiogenesis and remodeling, as well as tissue repair and regeneration.

[0153]

[0167] CXCL7 belongs to the CXC chemokine family and is a potent chemoattractant and neutrophil activator. It has been shown to stimulate various cellular processes, including DNA synthesis, mitosis, glycolysis, intracellular cAMP accumulation, prostaglandin E2 secretion, and the synthesis of hyaluronic acid and sulfated glycosaminoglycans.

[0154]

[0168] CD163 is a member of the cysteine-rich (SRCR) scavenger receptor superfamily and is primarily expressed in monocytes and macrophages. It functions as an acute-phase regulatory receptor involved in macrophage-mediated clearance and endocytosis of hemoglobin / haptoglobin complexes, thereby potentially protecting tissues from free hemoglobin-mediated oxidative damage. Soluble CD163 (sCD163) is released from the cell surface by proteolysis following oxidative stress or inflammatory stimuli. sCD163 is considered a biomarker of macrophage activation in several inflammatory diseases, including sepsis, inflammatory bowel disease, and influenza-associated encephalopathy. See Etzerodt et al., Scientific Reports, 2017; 7:40286.

[0155]

[0169] In some embodiments, the expression level of the biomarker increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared with a control sample. In some embodiments, the expression level of the biomarker in biological samples is approximately 1.1 times, approximately 1.2 times, approximately 1.3 times, approximately 1.4 times, approximately 1.5 times, approximately 1.6 times, approximately 1.7 times, approximately 1.8 times, approximately 1.9 times, approximately 2.0 times, approximately 2.1 times, approximately 2.2 times, approximately 2.3 times, approximately 2.4 times, approximately 2.5 times, approximately 2.6 times, approximately 2.7 times, approximately 2.8 times, approximately 2.9 times, approximately 3.0 times, approximately 3.1 times, approximately 3.2 times, approximately 3.3 times, approximately 3.4 times, approximately 3.5 times, approximately 3.6 times, approximately 3.7 times, approximately 3.8 times, approximately 3.9 times, approximately 4.0 times, approximately 4.1 times, approximately 4.2 times, approximately 4.3 times, approximately 4.4 times, approximately 4.5 times, approximately 4.6 times, approximately 4.7 times, approximately 4.8 times, approximately 4.9 times, approximately 5.0 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, approximately 20 times, approximately 30 times, approximately 40 times, approximately 50 times, approximately 60 times, approximately 70 times, approximately 80 times, approximately 90 times, approximately 100 times, approximately 500 times, approximately 1,000 times, and beyond. In some embodiments, the expression level of the biomarker in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to a control sample. In some embodiments, the increase in the expression level of the biomarker in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, such as interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis.In some embodiments, the biomarker is selected from the group consisting of MYDGF, soluble RAGE, phosphorylated SMAD2 (pSMAD2), pSMAD3, PDGFRβ, pPDGFRβ, PROF1, CALM2, CALR, PPIA, EIF5A, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR.

[0156]

[0170] In some embodiments, the expression level of MYDGF increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivative) compared to a control sample. In some embodiments, the expression level of MYDGF in biological samples is approximately 1.1 times, approximately 1.2 times, approximately 1.3 times, approximately 1.4 times, approximately 1.5 times, approximately 1.6 times, approximately 1.7 times, approximately 1.8 times, approximately 1.9 times, approximately 2.0 times, approximately 2.1 times, approximately 2.2 times, approximately 2.3 times, approximately 2.4 times, approximately 2.5 times, approximately 2.6 times, approximately 2.7 times, approximately 2.8 times, approximately 2.9 times, approximately 3.0 times, approximately 3.1 times, approximately 3.2 times, approximately 3.3 times, approximately 3.4 times, approximately 3 0.5 times, approximately 3.6 times, approximately 3.7 times, approximately 3.8 times, approximately 3.9 times, approximately 4.0 times, approximately 4.1 times, approximately 4.2 times, approximately 4.3 times, approximately 4.4 times, approximately 4.5 times, approximately 4.6 times, approximately 4.7 times, approximately 4.8 times, approximately 4.9 times, approximately 5.0 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, approximately 20 times, approximately 30 times, approximately 40 times, approximately 50 times, approximately 60 times, approximately 70 times, approximately 80 times, approximately 90 times, approximately 100 times, approximately 500 times, approximately 1,000 times, and beyond. In some embodiments, the expression level of MYDGF in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to the control sample. In some embodiments, the increase in the expression level of MYDGF in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis.

[0157]

[0171] In some embodiments, the expression level of soluble RAGE increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivative) compared to a control sample. In some embodiments, the expression level of soluble RAGE in biological samples is approximately 1.1 times, approximately 1.2 times, approximately 1.3 times, approximately 1.4 times, approximately 1.5 times, approximately 1.6 times, approximately 1.7 times, approximately 1.8 times, approximately 1.9 times, approximately 2.0 times, approximately 2.1 times, approximately 2.2 times, approximately 2.3 times, approximately 2.4 times, approximately 2.5 times, approximately 2.6 times, approximately 2.7 times, approximately 2.8 times, approximately 2.9 times, approximately 3.0 times, approximately 3.1 times, approximately 3.2 times, approximately 3.3 times, approximately 3.4 times, approximately 3.5 times, approximately 3.6 times, approximately 3.7 times, approximately 3.8 times, approximately 3.9 times, approximately 4.0 times, approximately 4.1 times, approximately 4.2 times, approximately 4.3 times, approximately 4.4 times, approximately 4.5 times, approximately 4.6 times, approximately 4.7 times, approximately 4.8 times, approximately 4.9 times, approximately 5.0 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, approximately 20 times, approximately 30 times, approximately 40 times, approximately 50 times, approximately 60 times, approximately 70 times, approximately 80 times, approximately 90 times, approximately 100 times, approximately 500 times, approximately 1,000 times, and beyond. In some embodiments, the expression level of soluble RAGE in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to the control sample. In some embodiments, the increase in the expression level of soluble RAGE in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis.

[0158]

[0172] In some embodiments, the expression level of prolifin increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivative) compared to a control sample. In some embodiments, the expression level of prolifin in biological samples is approximately 1.1 times, 1.2 times, 1.3 times, 1.4 times, 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2.0 times, 2.1 times, 2.2 times, 2.3 times, 2.4 times, 2.5 times, 2.6 times, 2.7 times, 2.8 times, 2.9 times, 3.0 times, 3.1 times, 3.2 times, 3.3 times, and 3 times compared to a control sample. 0.4 times, approximately 3.5 times, approximately 3.6 times, approximately 3.7 times, approximately 3.8 times, approximately 3.9 times, approximately 4.0 times, approximately 4.1 times, approximately 4.2 times, approximately 4.3 times, approximately 4.4 times, approximately 4.5 times, approximately 4.6 times, approximately 4.7 times, approximately 4.8 times, approximately 4.9 times, approximately 5.0 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, approximately 20 times, approximately 30 times, approximately 40 times, approximately 50 times, approximately 60 times, approximately 70 times, approximately 80 times, approximately 90 times, approximately 100 times, approximately 500 times, approximately 1,000 times, and beyond. In some embodiments, the expression level of prolifin in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to the control sample. In some embodiments, the increase in the expression level of prolifin in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, such as interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis.

[0159]

[0173] In some embodiments, the expression level of calmodulin-2 increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivative) compared to a control sample. In some embodiments, the expression level of calmodulin-2 in biological samples is approximately 1.1 times, approximately 1.2 times, approximately 1.3 times, approximately 1.4 times, approximately 1.5 times, approximately 1.6 times, approximately 1.7 times, approximately 1.8 times, approximately 1.9 times, approximately 2.0 times, approximately 2.1 times, approximately 2.2 times, approximately 2.3 times, approximately 2.4 times, approximately 2.5 times, approximately 2.6 times, approximately 2.7 times, approximately 2.8 times, approximately 2.9 times, approximately 3.0 times, approximately 3.1 times, approximately 3.2 times, approximately 3.3 times, approximately 3.4 times, It increases by approximately 3.5 times, 3.6 times, 3.7 times, 3.8 times, 3.9 times, 4.0 times, 4.1 times, 4.2 times, 4.3 times, 4.4 times, 4.5 times, 4.6 times, 4.7 times, 4.8 times, 4.9 times, 5.0 times, 6 times, 7 times, 8 times, 9 times, 10 times, 20 times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times, 100 times, 500 times, 1,000 times, and beyond. In some embodiments, the expression level of calmodulin-2 in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to the control sample. In some embodiments, the increase in the expression level of calmodulin-2 in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, such as interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis.

[0160]

[0174] In some embodiments, the expression level of calreticulin increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivative) compared to a control sample. In some embodiments, the expression level of calreticulin in biological samples is approximately 1.1 times, approximately 1.2 times, approximately 1.3 times, approximately 1.4 times, approximately 1.5 times, approximately 1.6 times, approximately 1.7 times, approximately 1.8 times, approximately 1.9 times, approximately 2.0 times, approximately 2.1 times, approximately 2.2 times, approximately 2.3 times, approximately 2.4 times, approximately 2.5 times, approximately 2.6 times, approximately 2.7 times, approximately 2.8 times, approximately 2.9 times, approximately 3.0 times, approximately 3.1 times, approximately 3.2 times, approximately 3.3 times, approximately 3.4 times, It increases by approximately 3.5 times, 3.6 times, 3.7 times, 3.8 times, 3.9 times, 4.0 times, 4.1 times, 4.2 times, 4.3 times, 4.4 times, 4.5 times, 4.6 times, 4.7 times, 4.8 times, 4.9 times, 5.0 times, 6 times, 7 times, 8 times, 9 times, 10 times, 20 times, 30 times, 40 times, 50 times, 60 times, 70 times, 80 times, 90 times, 100 times, 500 times, 1,000 times, and beyond. In some embodiments, the expression level of calreticulin in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to a control sample. In some embodiments, the increase in the expression level of calreticulin in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, such as interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis (IPF). In some embodiments, the biological sample is fibroblasts. In some embodiments, the biological sample is IPF fibroblasts.

[0161]

[0175] In some embodiments, the expression level of PPIA increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of PPIA in biological samples is approximately 1.1 times, approximately 1.2 times, approximately 1.3 times, approximately 1.4 times, approximately 1.5 times, approximately 1.6 times, approximately 1.7 times, approximately 1.8 times, approximately 1.9 times, approximately 2.0 times, approximately 2.1 times, approximately 2.2 times, approximately 2.3 times, approximately 2.4 times, approximately 2.5 times, approximately 2.6 times, approximately 2.7 times, approximately 2.8 times, approximately 2.9 times, approximately 3.0 times, approximately 3.1 times, approximately 3.2 times, approximately 3.3 times, approximately 3.4 times, approximately 3 0.5 times, approximately 3.6 times, approximately 3.7 times, approximately 3.8 times, approximately 3.9 times, approximately 4.0 times, approximately 4.1 times, approximately 4.2 times, approximately 4.3 times, approximately 4.4 times, approximately 4.5 times, approximately 4.6 times, approximately 4.7 times, approximately 4.8 times, approximately 4.9 times, approximately 5.0 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, approximately 20 times, approximately 30 times, approximately 40 times, approximately 50 times, approximately 60 times, approximately 70 times, approximately 80 times, approximately 90 times, approximately 100 times, approximately 500 times, approximately 1,000 times, and beyond. In some embodiments, the expression level of PPIA in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to the control sample. In some embodiments, the increase in the expression level of PPIA in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis.

[0162]

[0176] In some embodiments, the expression level of EIF5A1 increases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivative) compared to a control sample. In some embodiments, the expression level of EIF5A1 in biological samples is approximately 1.1 times, approximately 1.2 times, approximately 1.3 times, approximately 1.4 times, approximately 1.5 times, approximately 1.6 times, approximately 1.7 times, approximately 1.8 times, approximately 1.9 times, approximately 2.0 times, approximately 2.1 times, approximately 2.2 times, approximately 2.3 times, approximately 2.4 times, approximately 2.5 times, approximately 2.6 times, approximately 2.7 times, approximately 2.8 times, approximately 2.9 times, approximately 3.0 times, approximately 3.1 times, approximately 3.2 times, approximately 3.3 times, approximately 3.4 times, approximately 3.5 times, approximately 3.6 times, approximately 3.7 times, approximately 3.8 times, approximately 3.9 times, approximately 4.0 times, approximately 4.1 times, approximately 4.2 times, approximately 4.3 times, approximately 4.4 times, approximately 4.5 times, approximately 4.6 times, approximately 4.7 times, approximately 4.8 times, approximately 4.9 times, approximately 5.0 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, approximately 20 times, approximately 30 times, approximately 40 times, approximately 50 times, approximately 60 times, approximately 70 times, approximately 80 times, approximately 90 times, approximately 100 times, approximately 500 times, approximately 1,000 times, and beyond. In some embodiments, the expression level of EIF5A1 in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to the control sample. In some embodiments, the increase in the expression level of EIF5A1 in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis.

[0163]

[0177] In some embodiments, LGALS7 expression levels increase in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to control samples. In some embodiments, LGALS7 expression levels in biological samples are approximately 1.1 times, 1.2 times, 1.3 times, 1.4 times, 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2.0 times, 2.1 times, 2.2 times, 2.3 times, 2.4 times, 2.5 times, 2.6 times, 2.7 times, 2.8 times, 2.9 times, 3.0 times, 3.1 times, 3.2 times, 3.3 times, 3.4 times, and approximately 3.5 times, approximately 3.6 times, approximately 3.7 times, approximately 3.8 times, approximately 3.9 times, approximately 4.0 times, approximately 4.1 times, approximately 4.2 times, approximately 4.3 times, approximately 4.4 times, approximately 4.5 times, approximately 4.6 times, approximately 4.7 times, approximately 4.8 times, approximately 4.9 times, approximately 5.0 times, approximately 6 times, approximately 7 times, approximately 8 times, approximately 9 times, approximately 10 times, approximately 20 times, approximately 30 times, approximately 40 times, approximately 50 times, approximately 60 times, approximately 70 times, approximately 80 times, approximately 90 times, approximately 100 times, approximately 500 times, approximately 1,000 times, and beyond. In some embodiments, the expression level of LGALS7 in the biological sample increases by approximately 110%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475%, 500%, 600%, 700%, 800%, 900%, 1000%, or more compared to the control sample. In some embodiments, the increase in the expression level of LGALS7 in the biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis. In some embodiments, biological samples are obtained from elderly subjects with a disease or disorder, such as interstitial lung disease.

[0164]

[0178] In some embodiments, the expression level of a biomarker decreases in a biological sample treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of a biomarker in a biological sample decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the decrease in the expression level of a biomarker in a biological sample indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis. In some embodiments, the biomarker is selected from the group consisting of MYDGF, soluble RAGE, phosphorylated SMAD2 (pSMAD2), pSMAD3, PDGFRβ, pPDGFRβ, PROF1, CALM2, CALR, PPIA, EIF5A, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR.

[0165]

[0179] In some embodiments, the expression levels of pSMAD2 and / or pSMAD3 are reduced in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression levels of pSMAD2 and / or pSMAD3 in biological samples are reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, an increase in the expression levels of pSMAD2 and / or pSMAD3 in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis.

[0166]

[0180] In some embodiments, the expression level of PDGFRβ decreases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of PDGFRβ in biological samples decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the decrease in the expression level of PDGFRβ in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis.

[0167]

[0181] In some embodiments, the expression level of pPDGFRβ decreases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of pPDGFRβ in biological samples decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the decrease in the expression level of pPDGFRβ in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis.

[0168]

[0182] In some embodiments, the expression level of calreticulin decreases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of calreticulin in biological samples decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, an increase in the expression level of calreticulin in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis (IPF). In some embodiments, the biological sample is epithelial cells. In some embodiments, the biological sample is IPF epithelial cells.

[0169]

[0183] In some embodiments, the expression level of p-mTOR decreases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of p-mTOR in biological samples decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the decrease in the expression level of p-mTOR in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis.

[0170]

[0184] In some embodiments, LGALS7 expression levels decrease in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, LGALS7 expression levels in biological samples decrease by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, an increase in LGALS7 expression levels in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis. In some embodiments, biological samples are obtained from elderly subjects with a disease or disorder, such as interstitial lung disease.

[0171]

[0185] In some embodiments, the expression level of IL-11 decreases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of IL-11 in biological samples decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the decrease in the expression level of IL-11 in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis. In some embodiments, biological samples are obtained from elderly subjects with a disease or disorder, such as interstitial lung disease.

[0172]

[0186] In some embodiments, the expression level of MMP-2 decreases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of MMP-2 in biological samples decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the decrease in the expression level of MMP-2 in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis. In some embodiments, biological samples are obtained from elderly subjects with a disease or disorder, such as interstitial lung disease.

[0173]

[0187] In some embodiments, the expression level of CXCL7 is reduced in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of CXCL7 in biological samples is reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the reduction in the expression level of CXCL7 in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis. In some embodiments, biological samples are obtained from elderly subjects with a disease or disorder, such as interstitial lung disease.

[0174]

[0188] In some embodiments, the expression level of SCD163 decreases in biological samples treated with a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) compared to a control sample. In some embodiments, the expression level of SCD163 in biological samples decreases by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, an increase in the expression level of SCD163 in biological samples indicates a favorable response to a therapeutic activator (e.g., caveolin-1 peptide or its derivatives). In some embodiments, biological samples are obtained from subjects with a disease or disorder, e.g., interstitial lung disease. In some embodiments, biological samples are obtained from subjects with idiopathic pulmonary fibrosis. In some embodiments, biological samples are obtained from elderly subjects with a disease or disorder, such as interstitial lung disease.

[0175]

[0189] In some embodiments, a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) reduces receptor tyrosine kinase (RTK)-related signaling pathways in a biological sample compared to a control sample. In some embodiments, the RTK-related signaling pathways are reduced by approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more compared to a control sample. In some embodiments, the reduction in RTK-related signaling pathways is a reduction in the expression levels of ALK, c-Jun, Herb2 / ErbB3, Stat5a, PI3Kp110a, SRC-1, and / or YAP. In some embodiments, the reduction in RTK-related signaling pathways is a reduction in the phosphorylation of ALK (p-ALK), c-Myc (pc-Myc), EGFR (p-EGFR), MEK1 / 2 (p-MEK-1 / 2), MAPK (p-MAPK), PDK1 (p-PDK1), PDGFRb (p-PDGFRβ), RafB (p-RafB), Ret (p-Ret), Stat5a (p-Stat5a), and / or SRC (p-SRC). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis. In some embodiments, the biological sample is fibroblasts obtained from a subject with a fibrous disease (e.g., idiopathic pulmonary fibrosis).

[0176]

[0190] In some embodiments, a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) reduces metabolic signaling pathways in a biological sample compared to a control sample. In some embodiments, metabolic signaling pathways are reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more, compared to a control sample. In some embodiments, the reduction in metabolic signaling pathways is a decrease in the expression levels of AMPKa, Deptor, LDHA, PFKFB3, and / or Raptor. In some embodiments, the reduction in metabolic signaling pathways is a decrease in the phosphorylation of AMPKa1 (p-AMPKa1), AMPKb1 (p-AMPKb1), mTOR (p-mTOR), Raptor (p-Raptor), and / or tuberine (p-tuberine). In some embodiments, the biological sample is obtained from a subject with a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject with idiopathic pulmonary fibrosis. In some embodiments, the biological sample is fibroblasts obtained from a subject with a fibrous disease (e.g., idiopathic pulmonary fibrosis).

[0177]

[0191] In some embodiments, a therapeutic activator (e.g., caveolin-1 peptide or its derivatives) reduces invasion-related molecules in the biological sample compared to a control sample. In some embodiments, invasion-related molecules are reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the reduction in invasion-related molecules is a reduction in the expression levels of TWIST2 and / or Wnt5ab.

[0178]

[0192] In some embodiments, a therapeutic activator (e.g., caveolin-1 peptide or its derivative) reduces histone deacetylase (HDAC) in a biological sample compared to a control sample. In some embodiments, HDAC is reduced by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more compared to a control sample. In some embodiments, the reduction in HDAC is a reduction in the expression levels of HDAC4 and / or HDAC6. In some embodiments, the biological sample is obtained from a subject having a disease or disorder, e.g., interstitial lung disease. In some embodiments, the biological sample is obtained from a subject having idiopathic pulmonary fibrosis. In some embodiments, the biological sample is fibroblasts obtained from a subject having a fibrous disease (e.g., idiopathic pulmonary fibrosis).

[0179]

[0193] In some embodiments, the methods provided herein reduce RTK-related signaling in subjects with fibrous disease. In some embodiments, the methods include administering a caveolin-1 peptide or a derivative thereof to subjects with fibrous disease. In some embodiments, the caveolin-1 peptide or a derivative thereof comprises one of the amino acid sequences of SEQ ID NOs: 1 to 110. In some embodiments, the caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 4.

[0180]

[0194] In some embodiments, the methods provided herein reduce RTK-related signaling in fibrous fibroblasts. In some embodiments, the methods include administering caveolin-1 peptide or a derivative thereof to fibrous fibroblasts. In some embodiments, caveolin-1 peptide or a derivative thereof does not affect RTK-related signaling in normal fibroblasts. In some embodiments, caveolin-1 peptide or a derivative thereof reduces RTK-related signaling in normal fibroblasts to a lower degree than that in fibrous fibroblasts. In some embodiments, caveolin-1 peptide or a derivative thereof comprises one of the amino acid sequences of SEQ ID NOs: 1 to 110. In some embodiments, caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 4.

[0181]

[0195] In some embodiments, the methods provided herein reduce metabolic signaling in subjects with fibrous disease. In some embodiments, the methods include administering a caveolin-1 peptide or a derivative thereof to subjects with fibrous disease. In some embodiments, the caveolin-1 peptide or a derivative thereof comprises one of the amino acid sequences of SEQ ID NOs: 1 to 110. In some embodiments, the caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, the caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 4.

[0182]

[0196] In some embodiments, the methods provided herein reduce metabolic signaling in fibrous fibroblasts. In some embodiments, the methods include administering caveolin-1 peptide or a derivative thereof to fibrous fibroblasts. In some embodiments, caveolin-1 peptide or a derivative thereof does not affect metabolic signaling in normal fibroblasts. In some embodiments, caveolin-1 peptide or a derivative thereof reduces metabolic signaling in normal fibroblasts to a lower degree than that in fibrous fibroblasts. In some embodiments, caveolin-1 peptide or a derivative thereof comprises one of the amino acid sequences of SEQ ID NOs: 1 to 110. In some embodiments, caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 3. In some embodiments, caveolin-1 peptide or a derivative thereof comprises the amino acid sequence of SEQ ID NO: 4.

[0183]

[0197] In some embodiments, one or more biomarkers are identified as being associated with a therapeutic agent in subjects with a disease or disorder, such as interstitial lung disease. In some embodiments, one or more biomarkers are identified as being associated with caveolin-1 therapy in subjects with idiopathic pulmonary fibrosis.

[0184]

[0198] In some embodiments, one or more biomarkers are used to determine or predict the optimal dose of a therapeutic agent in subjects with a disease or disorder, such as interstitial lung disease. In some embodiments, one or more biomarkers are used to determine or predict the optimal dose of caveolin-1 peptide or its derivatives (e.g., CSP-7 or Var55) in subjects with idiopathic pulmonary fibrosis.

[0185] subject

[0199] As used herein, the term “subject” refers to a mammalian subject. In some embodiments, the subject is a human, a companion animal, a non-domesticated animal, or a zoo animal. In some embodiments, the subject is a human, a mouse, a rat, a rabbit, a hamster, a guinea pig, a dog, a cat, a cow, a pig, a sheep, a goat, a horse, or a monkey. In some embodiments, the subject is a human.

[0186]

[0200] In some embodiments, the subject has a disease or disorder. In some embodiments, the subject having a disease or disorder is treated with a therapeutic activator, e.g., caveolin-1 peptide or its derivatives. In some embodiments, the biomarkers described herein are associated with a therapeutic activator, e.g., caveolin-1 peptide or its derivatives, in a subject having a disease or disorder. Examples of diseases or disorders include autoimmune diseases (e.g., diabetes, lupus, multiple sclerosis, psoriasis, rheumatoid arthritis); inflammatory diseases (e.g., arthritis, pelvic inflammatory disease); infectious diseases (e.g., viral infections (e.g., HIV, HCV, RSV, COVID-19)), bacterial infections, fungal infections, sepsis); neurological disorders (e.g., Alzheimer's disease, Huntington's disease; autism; Duchenne muscular dystrophy); cardiovascular disorders (e.g., atherosclerosis, hypercholesterolemia, thrombosis, coagulation disorders, neovascular disorders such as macular degeneration); proliferative This includes, but is not limited to, diseases (e.g., cancer, benign neoplasms); respiratory diseases (e.g., chronic obstructive pulmonary disease); digestive diseases (e.g., inflammatory bowel disease, ulcers); musculoskeletal diseases (e.g., fibromyalgia, arthritis); endocrine, metabolic, and nutritional disorders (e.g., diabetes mellitus, osteoporosis); urinary tract diseases (e.g., kidney disease); mental disorders (e.g., depression, schizophrenia); skin diseases (e.g., wounds, eczema); blood and lymphatic disorders (e.g., anemia, hemophilia); fibrous diseases (e.g., idiopathic pulmonary fibrosis); or eye diseases (e.g., glaucoma, diabetic retinopathy).

[0187]

[0201] In some embodiments, the subjects have fibrous diseases. In some embodiments, subjects having fibrous diseases are treated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof. In some embodiments, the biomarkers described herein are associated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof, in subjects having fibrous diseases. Fibrous diseases include those of the bone marrow, gallbladder, blood vessels, heart, joints, kidneys, liver, lungs, muscles, pancreas, penis, skin, soft tissues, eyes, adrenal glands, thyroid gland and / or uterus. Examples of fibrotic diseases include pulmonary fibrosis, acute interstitial pneumonia, nonspecific interstitial pneumonia, lymphocytic interstitial pneumonia, asthma, idiopathic interstitial fibrosis, interstitial lung disease, interstitial pneumonia, cystic fibrosis, idiopathic organizing pneumonia, desquamative interstitial pneumonia, diffuse parenchymal lung disease, respiratory bronchiolitis, subepithelial fibrosis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, endocardial fibrosis, pancreatic fibrosis, chronic pancreatitis, liver cirrhosis, chronic kidney disease, gallbladder cirrhosis, nephrosclerosis, and glomerulosclerosis. Systemic nephritis, arteriosclerosis, atherosclerosis, restenosis, macular degeneration, ocular scarring, cataracts, retinal and vitreoretinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloid scarring, hypertrophic scarring, colloid and hypertrophic scarring, postoperative scarring, abnormal wound healing, peritoneal fibrosis, retroperitoneal fibrosis, chronic obstructive pulmonary disease, postoperative myomas, diabetic nephropathy, nephrogenic systemic fibrosis, gynecological cancers, chronic myeloproliferative disorders, myelofibrosis, bone Myeloproliferative syndrome, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease (e.g., Crohn's disease and collagen colitis), alcoholic steatohepatitis, alcoholic fatty liver disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, rheumatoid pannus formation in rheumatoid arthritis or osteoarthritis, fibrosis as a result of graft-versus-host disease, organ transplant fibrosis, anthrofibrosis, non-alcoholic steatohepatitis, Alport syndrome, chronic COVID syndrome, glial scar formation in HIV, Peyronie's disease, associated cognitive and motor disorders and cavernous encephalopathy, gingival hyperplasia secondary to drugs and fibrocystic diseases, Dupuytren's contracture, Morphea, endometriosis, uterine fibroids, fibromyalgia, multifocal fibrosclerosis, progressive ossifying fibrodysplasia, osteoporosis, or osteosclerosis.

[0188]

[0202] In some embodiments, the subjects have a lung disease. In some embodiments, subjects having a lung disease are treated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof. In some embodiments, the biomarkers described herein are associated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof, in subjects having a lung disease. Examples of lung diseases include, but are not limited to, pulmonary fibrosis, interstitial lung disease, chronic obstructive pulmonary disease (COPD), acute lung injury, acute respiratory distress syndrome, chronic bronchitis, emphysema, lung cancer, cystic fibrosis, bronchiectasis, pneumonia, pleural effusion, tuberculosis, pulmonary hypertension, collagenous vascular lung disease (e.g., due to lupus, scleroderma, or mixed connective tissue disease), and asthma. Lung diseases may be caused by smoking, exposure to irritants (e.g., asbestos and silica), allergies, genetics, or unknown causes.

[0189]

[0203] In some embodiments, the subject has interstitial lung disease (ILD). In some embodiments, the subject having ILD is treated with a therapeutic agent, such as caveolin-1 peptide or its derivatives. In some embodiments, the biomarkers described herein are associated with the therapeutic agent, such as caveolin-1 peptide or its derivatives, in the subject having ILD. Examples of ILD include, but are not limited to, idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, and pulmonary sarcoidosis. ILD may be associated with connective tissue diseases selected from systemic sclerosis, polymyositis, systemic lupus erythematosus, or rheumatoid arthritis. ILD can also be drug-induced, such as by antibiotics, chemotherapeutic agents, antiarrhythmics, statins, etc. ILD can also result from viral infections (e.g., COVID-19), bacterial infections, or tuberculosis. ILD may be associated with environmental or occupational exposure to silica, asbestos, talc, hard metals, inorganic or organic dust, radiation, gases / fumes, etc.

[0190]

[0204] In some embodiments, the subjects have pulmonary fibrosis. In some embodiments, subjects having pulmonary fibrosis are treated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof. In some embodiments, the biomarkers described herein are associated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof, in subjects having pulmonary fibrosis. Examples of pulmonary fibrosis include, but are not limited to, familial pulmonary fibrosis, interstitial lung disease, idiopathic pulmonary fibrosis, idiopathic nonspecific interstitial pneumonia, conventional interstitial lung disease, idiopathic organizing pneumonia, pulmonary sarcoidosis, fibrotic alveolitis, cystic fibrosis, COPD, adult respiratory distress syndrome, and emphysema. Pulmonary fibrosis can be caused by many conditions, including chronic inflammatory processes (e.g., sarcoidosis and Wegener's granulomatosis), infections (e.g., COVID-19), environmental factors (e.g., exposure to asbestos, silica, hard metals, talc, and certain gases), exposure to ionizing radiation (e.g., radiotherapy to treat thoracic tumors), chronic conditions (e.g., lupus, rheumatoid arthritis), and even certain medications.

[0191]

[0205] In some embodiments, the subjects have idiopathic pulmonary fibrosis. In some embodiments, subjects having idiopathic pulmonary fibrosis are treated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof. In some embodiments, the biomarkers described herein are associated with a therapeutic agent, such as caveolin-1 peptide or a derivative thereof, in subjects having idiopathic pulmonary fibrosis.

[0192] Biological samples

[0206] As used herein, the term “biological sample” refers to a sample obtained from a subject. Biological samples may be obtained from a subject using methods known to those skilled in the art. Examples of biological samples include, but are not limited to, bodily fluids, organs, tissues, and cells isolated from mammals. In some embodiments, a biological sample is a section of a biological sample, e.g., a section of an organ or tissue. In some embodiments, a biological sample is an extract from a biological sample, e.g., an antigen from a biological fluid. Methods for collecting, handling, and processing biological samples obtained from a subject are well known in the art.

[0193]

[0207] In some embodiments, biological samples are obtained from mammals (e.g., rats, mice, rabbits, dogs, cats, cattle, horses, donkeys, guinea pigs, monkeys, or humans). In some embodiments, biological samples are obtained from primates (e.g., chimpanzees or humans). In some embodiments, biological samples are obtained from humans. In some embodiments, biological samples are obtained from humans with a disease or disorder, such as interstitial lung disease. In some embodiments, biological samples are obtained from humans with idiopathic pulmonary fibrosis. In some embodiments, biological samples are obtained from humans without evidence of disease or disorder (e.g., control samples from healthy human subjects).

[0194]

[0208] In some embodiments, the biological sample is a body fluid. Examples of body fluids include, but are not limited to, blood, plasma, serum, urine, peritoneal fluid, cerebrospinal fluid, amniotic fluid, bronchoalveolar lavage fluid, lymph, saliva, pleural fluid, or interstitial fluid. In some embodiments, the biological sample is plasma. In some embodiments, the biological sample is serum. In some embodiments, the biological sample is bronchoalveolar lavage fluid.

[0195]

[0209] In some embodiments, the biological sample is a tissue or cell sample. The source of the tissue or cell sample may be fresh, frozen, and / or preserved organ or tissue sample or solid tissue from a biopsy or aspirate; or cells from any stage of the subject's pregnancy or development. The biological sample may also be cultured primary cells or cell lines derived from a subject with a disease or disorder, such as interstitial lung disease.

[0196]

[0210] In some embodiments, the biological sample is a tissue sample obtained from a subject. In some embodiments, the biological sample is lung tissue, skin tissue, heart tissue, liver tissue, gastrointestinal tissue, pancreatic tissue, bone tissue, nerve tissue, adipose tissue, cartilage tissue, connective tissue, muscle tissue, epithelial tissue, or eye tissue. In some embodiments, the biological sample is lung tissue from a subject with idiopathic pulmonary fibrosis.

[0197]

[0211] In some embodiments, the biological sample is a cell sample obtained from a subject. In some embodiments, the biological sample includes fibroblasts, conditionally reprogrammed cells, type I alveolar epithelial cells, type II alveolar epithelial cells, basal cells, Clara cells, ciliated cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells (BASCs), alveolar macrophages, ionocytes, pericytes, mesothelial cells, erythrocytes, leukocytes, peripheral blood mononuclear cells, B cells, plasma cells, innate lymphocytes, T cells (e.g., CD8+ T cells, CD4+ T cells, regulatory T cells), macrophages, and non-classical monocellular cells. The cells are globules, monocytes, neutrophils, eosinophils, dendritic cells, mast cells, NK cells, stem cells, cholangiocarcinoma cells, hepatocytes, Kupffer cells, keratinocytes, melanocytes, pyramidal cells, rod cells, pigment cells, Langerhan cells, cardiomyocytes, smooth muscle cells, myofibroblasts, stromal cells, osteocytes, adipocytes, muscle cells, epithelial cells, endocrine cells, germ cells, vascular cells, nerve cells, glial cells, trophoblast cells, mesenchymal cells, cancer cells (including cancer cell lines), skin cells, nerve cells, or pancreatic cells. In some embodiments, the biological sample is fibroblasts from a subject with idiopathic pulmonary fibrosis. In some embodiments, the biological sample is basal-like cells from a subject with idiopathic pulmonary fibrosis. In some embodiments, the biological sample is conditionally reprogrammed cells from a subject with idiopathic pulmonary fibrosis. In some embodiments, the biological sample is epithelial cells from a subject with idiopathic pulmonary fibrosis.

[0198]

[0212] In some embodiments, the biological sample is the supernatant from cells or tissues cultured in vitro or ex vivo. In some embodiments, the supernatant is from cultured conditionally reprogrammed cells obtained from subjects with idiopathic pulmonary fibrosis. In some embodiments, the supernatant is from cultured lung tissue from subjects with idiopathic pulmonary fibrosis. In some embodiments, the supernatant is from cultured fibroblasts from subjects with idiopathic pulmonary fibrosis.

[0199]

[0213] In some embodiments, the biological sample is a control sample. As used herein, the term “control sample” refers to a biological sample obtained from a subject or population that has not been treated with the therapeutic activator described herein (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the control sample is used to determine or identify the expression level of a biomarker in a biological sample obtained from a subject treated with the therapeutic activator described herein (e.g., caveolin-1 peptide or its derivatives). In some embodiments, the control sample is a biological sample obtained from a subject or population of subjects having a disease or disorder, e.g., interstitial lung disease. In some embodiments, the control sample is a biological sample obtained from a healthy subject. In some embodiments, the control sample is a biological sample obtained from a subject before treatment with the therapeutic activator. In some embodiments, the control sample is a biological sample obtained from a subject before treatment with caveolin-1 peptide or its derivatives. In some embodiments, the control sample is a biological sample obtained from a subject before treatment with CSP-7 or Var55.

[0200]

[0214] In some embodiments, the biological sample contains compounds that do not naturally mix with the biological sample. Examples of such compounds include, but are not limited to, preservatives, anticoagulants, buffers, fixatives, nutrients, and antibiotics.

[0201] Use of biomarkers in caveolin-1 peptide therapy

[0215] In some embodiments, the present disclosure provides a method for treating a subject with caveolin-1 peptide therapy, wherein the subject suffers from fibrosis, and the method comprises (a) obtaining a biological sample from the subject or the step of obtaining one; (b) treating cells of the biological sample with caveolin-1 peptide or a derivative thereof or the step of having treated cells with such a sample; (c) measuring the expression level of a biomarker in the cells; and (d) comparing the expression level of the biomarker with that of a control sample; the biomarkers include bone marrow-derived growth factor (MYDGF), soluble RAGE, phosphorylated mothers... The antidecapentaplesic homolog 2 / 3 (pSMAD2 / 3), platelet-derived growth factor receptor beta (PDGFRβ), galectin-7 (LGALS7), interleukin-11 (IL-11), matrix metalloproteinase-2 (MMP-2), chemokine ligand 7 (CXCL-7), soluble CD163, phosphorylated mTOR (p-mTOR), phosphorylated PDGFRβ (pPDGFRβ), prolifin (PROF1), calmodulin 2 (CALM2), calreticulin (CALR), peptidyl-prolyl cis-trans isomerase A (PPIA), or eukaryotic translation initiation factor 5A (EIF5A1);

[0216] If the expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 are increased, administer caveolin-1 peptide or its derivatives to the target; or

[0217] If the expression levels of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, soluble CD163, or pSMAD2 / 3 are decreased, administer caveolin-1 peptide or its derivatives.

[0202]

[0218] In some embodiments, the method of administering the caveolin-1 peptide or its derivatives is as described herein (e.g., identity of the Cav-1 peptide, administration, route of administration, target, etc.).

[0203]

[0219] In some embodiments, the biological sample, cell type, biomarker measurement, and / or level of biomarker expression change are as described herein.

[0204]

[0220] In some embodiments, steps (b) to (d) of the method are repeated once or more times.

[0205]

[0221] In some embodiments, fibrosis is interstitial lung disease, hepatic fibrosis, renal fibrosis, dermatofibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataracts, retina and vitreoretinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloids and hypertrophic scars, peritoneal fibrous disease, chronic obstructive pulmonary disease, postoperative fibroids, diabetic nephropathy, gynecological cancers, myeloproliferative syndromes, myeloid leukemia, myelodysplastic syndromes, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome. In some embodiments, interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar injury, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease. In some embodiments, interstitial lung disease is idiopathic pulmonary fibrosis.

[0206]

[0222] The method according to claim 1, wherein the control sample is cells obtained from a subject before treatment with caveolin-1 peptide or a derivative thereof.

[0207]

[0223] In some embodiments, the methods provided herein are used to identify changes in the expression levels of biomarkers associated with a therapeutic agent in subjects having a disease or disorder. In some embodiments, the methods include (a) treating a biological sample from a subject with the therapeutic agent; (b) measuring the expression level of the biomarker in the biological sample; and (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample. In some embodiments, the therapeutic agent modulates the expression level of the biomarker in the biological sample. In some embodiments, the biomarker is selected from the group consisting of pSMAD2 / 3, PDGFRβ, pPDGFRβ, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, and / or soluble RAGE are associated with therapeutic activators in subjects with disease or impairment. In some embodiments, decreased expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 are associated with therapeutic activators in subjects with disease or impairment.

[0208]

[0224] In some embodiments, the methods provided herein are used to identify changes in the expression levels of biomarkers associated with caveolin-1 therapy in subjects with interstitial lung disease. In some embodiments, the methods include (a) treating cells from subjects with interstitial lung disease with a caveolin-1 peptide or a derivative thereof; (b) measuring the expression level of the biomarker in the cells; and (c) comparing the expression level of the biomarker in the cells with the expression level of the biomarker in a control sample. In some embodiments, the caveolin-1 peptide or a derivative thereof modulates the expression level of the biomarker in the cells. In some embodiments, the biomarker is selected from the group consisting of pSMAD2 / 3, PDGFRβ, pPDGFRβ, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, and / or soluble RAGE are associated with caveolin-1 peptides or their derivatives in subjects with disease or impairment. In some embodiments, decreased expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 are associated with caveolin-1 peptides or their derivatives in subjects with disease or impairment. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0209]

[0225] In some embodiments, the methods provided herein are used to identify changes in the expression levels of biomarkers associated with CSP-7 treatment in subjects with interstitial lung disease. In some embodiments, the methods include (a) treating cells from subjects with interstitial lung disease with CSP-7; (b) measuring the expression levels of the biomarkers in the cells; and (c) comparing the expression levels of the biomarkers in the cells with the expression levels of the biomarkers in a control sample. In some embodiments, CSP-7 modulates the expression levels of the biomarkers in the cells. In some embodiments, the biomarkers are selected from the group consisting of pSMAD2 / 3, PDGFRβ, pPDGFRβ, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, and / or soluble RAGE are associated with CSP-7 in subjects with interstitial lung disease. In some embodiments, decreased expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 are associated with CSP-7 in subjects with interstitial lung disease. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0210]

[0226] In some embodiments, the methods provided herein are used to identify changes in the expression levels of biomarkers associated with Var55 treatment in subjects with interstitial lung disease. In some embodiments, the methods include (a) treating cells from subjects with interstitial lung disease with Var55; (b) measuring the expression levels of the biomarkers in the cells; and (c) comparing the expression levels of the biomarkers in the cells with the expression levels of the biomarkers in a control sample. In some embodiments, Var55 modulates the expression levels of biomarkers in the cells. In some embodiments, the biomarker is selected from the group consisting of pSMAD2 / 3, PDGFRβ, pPDGFRβ, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, and / or soluble RAGE are associated with Var55 in subjects with interstitial lung disease. In some embodiments, decreased expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 are associated with Var55 in subjects with interstitial lung disease. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0211]

[0227] In some embodiments, the methods provided herein are used to predict or determine the efficacy of a therapeutic agent in a subject having a disease or disorder. In some embodiments, the methods include (a) treating a biological sample from the subject with the therapeutic agent; (b) measuring the expression level of a biomarker in the biological sample; and (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE indicate a favorable response to the therapeutic activator. In some embodiments, decreased expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicate a favorable response to the therapeutic activator.

[0212]

[0228] In some embodiments, the methods provided herein are used to predict or determine the efficacy of caveolin-1 peptide or its derivatives in subjects with interstitial lung disease. In some embodiments, the methods include (a) treating a biological sample from a subject with interstitial lung disease with caveolin-1 peptide or its derivatives; (b) measuring the expression level of a biomarker in the biological sample; and (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE indicate a favorable response to caveolin-1 peptide or its derivatives. In some embodiments, decreased expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicate a favorable response to caveolin-1 peptide or its derivatives. In some embodiments, the biological sample is pulmonary fibroblasts or basal-like cells obtained from a subject with interstitial lung disease. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0213]

[0229] In some embodiments, the methods provided herein are used to predict or determine the efficacy of CSP-7 in subjects with interstitial lung disease. In some embodiments, the methods include (a) treating a biological sample from a subject with CSP-7; (b) measuring the expression level of a biomarker in the biological sample; and (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE indicate a favorable response to CSP-7. In some embodiments, decreased expression levels of PDGFRβ, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, pSMAD2 / 3, or CALR indicate a favorable response to CSP-7. In some embodiments, the biological sample is pulmonary fibroblasts or basal-like cells obtained from a subject with interstitial lung disease. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0214]

[0230] In some embodiments, the methods provided herein are used to predict or determine the efficacy of Var55 in subjects with interstitial lung disease. In some embodiments, the methods include (a) treating a biological sample from a subject with Var55; (b) measuring the expression level of a biomarker in the biological sample; and (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE indicate a favorable response to Var55. In some embodiments, decreased expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicate a favorable response to Var55. In some embodiments, the biological sample is pulmonary fibroblasts or basal-like cells obtained from a subject with interstitial lung disease. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0215]

[0231] In some embodiments, the methods provided herein are used to determine the optimal dose of a therapeutic agent in a subject with a disease or disorder. In some embodiments, the method for determining the optimal dose of a therapeutic agent in a subject with a disease or disorder includes (a) obtaining a biological sample from the subject before treatment with the therapeutic agent; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; and (d) determining the optimal dose of the therapeutic agent to be administered to the subject based on the expression level of the biomarker in step (c). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ.

[0216]

[0232] In some embodiments, the methods provided herein are used to determine the optimal dose of caveolin-1 peptide or its derivatives in subjects with interstitial lung disease. In some embodiments, the method for determining the optimal dose of caveolin-1 peptide or its derivatives in subjects with interstitial lung disease includes (a) obtaining a biological sample from the subject before treatment with the caveolin-1 peptide or its derivatives; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; and (d) determining the optimal dose of caveolin-1 peptide or its derivatives to be administered to the subject based on the expression level of the biomarker in step (c). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0217]

[0233] In some embodiments, the methods provided herein are used to determine the optimal dose of CSP-7 in a subject having interstitial lung disease. In some embodiments, the method for determining the optimal dose of CSP-7 in a subject having interstitial lung disease includes (a) obtaining a biological sample from the subject before treatment with CSP-7; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; and (d) determining the optimal dose of CSP-7 to be administered to the subject based on the expression level of the biomarker in step (c). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0218]

[0234] In some embodiments, the methods provided herein are used to determine the optimal dose of Var55 in a subject having interstitial lung disease. In some embodiments, the method for determining the optimal dose of Var55 in a subject having interstitial lung disease includes (a) obtaining a biological sample from the subject before treatment with Var55; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; and (d) determining the optimal dose of Var55 to be administered to the subject based on the expression level of the biomarker in step (c). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, pSMAD2 / 3, MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0219]

[0235] In some embodiments of any one of the methods provided herein, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, pSMAD2 / 3, MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments of any one of the methods provided herein, the biomarker is selected from the group consisting of pSMAD2 / 3, PDGFRβ, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS1, ANXA5, FABP5, GSTP1, ENO1, and YWHAG.

[0220]

[0236] In some embodiments, one or more biological samples are collected from a subject with a disease or disorder after treatment with a therapeutic agent. In some embodiments, the expression level of a biomarker in the biological sample is measured and compared to a control sample (e.g., a biological sample obtained from the subject before treatment with the therapeutic agent). In some embodiments, the biomarker expression level is used to determine the efficacy of the therapeutic agent in the subject. In some embodiments, the biomarker expression level is used to determine the optimal dose of the therapeutic agent in the subject. In some embodiments, the biomarker expression level is used to monitor the administration of the therapeutic agent. In some embodiments, the therapeutic agent is a caveolin-1 peptide or a derivative thereof, e.g., CSP-7 or Var55.

[0221]

[0237] In some embodiments, the first and second biological samples are obtained from subjects having a disease or disorder, such as interstitial lung disease. In some embodiments, the first biological sample is obtained from the subject before treatment with the therapeutic agent. In some embodiments, the second biological sample is obtained from the subject after treatment with the therapeutic agent. In some embodiments, one or more additional biological samples are obtained from the subject after treatment with the therapeutic agent. In some embodiments, the second biological sample is obtained from the subject about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 18 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 1 year after treatment with the therapeutic agent. In some embodiments, one or more additional biological samples are obtained from the subject about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 18 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 6 months, or 1 year after treatment with the therapeutic activator. In some embodiments, the therapeutic activator is a caveolin-1 peptide or a derivative thereof, such as CSP-7 or Var55.

[0222]

[0238] The methods provided herein can be used to predict or determine the efficacy of a therapeutic agent in a subject with a disease or disorder. In some embodiments, the method for predicting or determining the efficacy of a therapeutic agent in a subject with a disease or disorder includes: (a) obtaining a first biological sample from the subject before treatment with the therapeutic agent; (b) administering the therapeutic agent to the subject; (c) obtaining a second biological sample from the subject after treatment with the therapeutic agent; (d) measuring the expression levels of biomarkers in the first and second biological samples; and (e) comparing the expression level of the biomarkers in the first biological sample with the expression level of the biomarkers in the second biological sample. In some embodiments, step (a) of the method further includes determining the expression level of the biomarkers in the first biological sample compared to a control sample before treatment with the therapeutic agent. In some embodiments, the expression level of the biomarkers in step (a) is used to determine the optimal dose of the therapeutic agent to be administered in step (b). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, an increase in the expression level of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE indicates a favorable response to the therapeutic agent. In some embodiments, a decrease in the expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicates a favorable response to the therapeutic activator.

[0223]

[0239] In some embodiments, the methods provided herein are used to predict or determine the efficacy of caveolin-1 peptide or its derivatives in subjects having interstitial lung disease. In some embodiments, the method for predicting or determining the efficacy of caveolin-1 peptide or its derivatives in subjects having interstitial lung disease includes: (a) obtaining a first biological sample from the subject before treatment with caveolin-1 peptide or its derivatives; (b) administering caveolin-1 peptide or its derivatives to the subject; (c) obtaining a second biological sample from the subject after treatment with caveolin-1 peptide or its derivatives; (d) measuring the expression levels of biomarkers in the first and second biological samples; and (e) comparing the expression level of biomarkers in the first biological sample with the expression level of biomarkers in the second biological sample. In some embodiments, step (a) of the method further includes determining the expression level of biomarkers in the first biological sample compared to a control sample before treatment with caveolin-1 peptide or its derivatives. In some embodiments, the expression level of the biomarker in step (a) is used to determine the optimal dose of the caveolin-1 peptide or its derivative administered in step (b). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE show a favorable response to caveolin-1 peptides or their derivatives.In some embodiments, a decrease in the expression levels of PDGFRβ, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or CALR indicates a favorable response to caveolin-1 peptide or its derivatives. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0224]

[0240] In some embodiments, the methods provided herein are used to predict or determine the efficacy of CSP-7 in subjects with interstitial lung disease. In some embodiments, the method for predicting or determining the efficacy of CSP-7 in subjects with interstitial lung disease includes (a) obtaining a first biological sample from the subject before treatment with CSP-7; (b) administering CSP-7 to the subject; (c) obtaining a second biological sample from the subject after treatment with CSP-7; (d) measuring the expression levels of biomarkers in the first and second biological samples; and (e) comparing the expression level of biomarkers in the first biological sample with the expression level of biomarkers in the second biological sample. In some embodiments, step (a) of the method further includes determining the expression level of biomarkers in the first biological sample compared to a control sample before treatment with CSP-7. In some embodiments, the expression level of biomarkers in step (a) is used to determine the optimal dose of CSP-7 to be administered in step (b). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is PDGFRβ. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE indicate a favorable response to CSP-7. In some embodiments, a decrease in the expression levels of PDGFRβ, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, pSMAD2 / 3, or CALR indicates a favorable response to CSP-7. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0225]

[0241] In some embodiments, the methods provided herein are used to predict or determine the efficacy of Var55 in subjects with interstitial lung disease. In some embodiments, the method for predicting or determining the efficacy of Var55 in subjects with interstitial lung disease includes (a) obtaining a first biological sample from the subject before treatment with Var55; (b) administering Var55 to the subject; (c) obtaining a second biological sample from the subject after treatment with Var55; (d) measuring the expression levels of biomarkers in the first and second biological samples; and (e) comparing the expression level of biomarkers in the first biological sample with the expression level of biomarkers in the second biological sample. In some embodiments, step (a) of the method further includes determining the expression level of biomarkers in the first biological sample compared to a control sample before treatment with Var55. In some embodiments, the expression level of biomarkers in step (a) is used to determine the optimal dose of Var55 to be administered in step (b). In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, increased expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, EIF5A1, LGALS7, or soluble RAGE indicate a favorable response to Var55. In some embodiments, a decrease in the expression levels of PDGFRβ, CALR, pPDGFRβ, p-mTOR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicates a favorable response to Var55. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0226]

[0242] In some embodiments, the methods provided herein are used to monitor the treatment of a therapeutic agent in a subject having a disease or disorder. In some embodiments, the method for monitoring the treatment of a therapeutic agent in a subject having a disease or disorder includes: (a) obtaining a first biological sample from the subject before treatment with the therapeutic agent; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; (d) administering the therapeutic agent to the subject; (e) obtaining a second biological sample from the subject after treatment with the therapeutic agent; (f) measuring the expression level of the biomarker in the second biological sample from the subject; and (g) comparing the expression level of the biomarker in the first biological sample with the expression level of the biomarker in the second biological sample. In some embodiments, if the expression level of the biomarker in the second biological sample is lower than the expression level of the biomarker in the first biological sample, the dose of the therapeutic agent is increased. In some embodiments, the dose of the therapeutic agent is increased if the expression level of the biomarker in the second biological sample is the same as the expression level of the biomarker in the first biological sample. In some embodiments, the dose of the therapeutic agent is maintained if the expression level of the biomarker in the second biological sample is higher than the expression level of the biomarker in the first biological sample. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ.

[0227]

[0243] In some embodiments, the methods provided herein are used to monitor treatment with caveolin-1 peptide or its derivatives in subjects having interstitial lung disease. In some embodiments, the method for monitoring treatment with caveolin-1 peptide or its derivatives in subjects having interstitial lung disease includes: (a) obtaining a first biological sample from the subject before treatment with caveolin-1 peptide or its derivatives; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; (d) administering the caveolin-1 peptide or its derivative to the subject; (e) obtaining a second biological sample from the subject after treatment with caveolin-1 peptide or its derivatives; (f) measuring the expression level of the biomarker in the second biological sample from the subject; and (g) comparing the expression level of the biomarker in the first biological sample with the expression level of the biomarker in the second biological sample. In some embodiments, if the expression level of the biomarker in the second biological sample is lower than the expression level of the biomarker in the first biological sample, the dose of caveolin-1 peptide or its derivative is increased. In some embodiments, if the expression level of the biomarker in the second biological sample is the same as the expression level of the biomarker in the first biological sample, the dose of caveolin-1 peptide or its derivative is increased. In some embodiments, if the expression level of the biomarker in the second biological sample is higher than the expression level of the biomarker in the first biological sample, the dose of caveolin-1 peptide or its derivative is maintained. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE.In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0228]

[0244] In some embodiments, the methods provided herein are used to monitor the treatment of CSP-7 in subjects having interstitial lung disease. In some embodiments, the method for monitoring the treatment of CSP-7 in subjects having interstitial lung disease includes: (a) obtaining a first biological sample from the subject before treatment with CSP-7; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; (d) administering CSP-7 to the subject; (e) obtaining a second biological sample from the subject after treatment with CSP-7; (f) measuring the expression level of the biomarker in the second biological sample from the subject; and (g) comparing the expression level of the biomarker in the first biological sample with the expression level of the biomarker in the second biological sample. In some embodiments, if the expression level of the biomarker in the second biological sample is lower than the expression level of the biomarker in the first biological sample, the dose of CSP-7 is increased. In some embodiments, the dose of CSP-7 is increased if the expression level of the biomarker in the second biological sample is the same as the expression level of the biomarker in the first biological sample. In some embodiments, the dose of CSP-7 is maintained if the expression level of the biomarker in the second biological sample is higher than the expression level of the biomarker in the first biological sample. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0229]

[0245] In some embodiments, the methods provided herein are used to monitor the treatment of Var55 in subjects having interstitial lung disease. In some embodiments, the method for monitoring the treatment of Var55 in subjects having interstitial lung disease includes: (a) obtaining a first biological sample from the subject before treatment with Var55; (b) measuring the expression level of a biomarker in the biological sample from the subject; (c) comparing the expression level of the biomarker in the biological sample with the expression level of the biomarker in a control sample; (d) administering Var55 to the subject; (e) obtaining a second biological sample from the subject after treatment with Var55; (f) measuring the expression level of the biomarker in the second biological sample from the subject; and (g) comparing the expression level of the biomarker in the first biological sample with the expression level of the biomarker in the second biological sample. In some embodiments, if the expression level of the biomarker in the second biological sample is lower than the expression level of the biomarker in the first biological sample, the dose of Var55 is increased. In some embodiments, the dose of Var55 is increased if the expression level of the biomarker in the second biological sample is the same as the expression level of the biomarker in the first biological sample. In some embodiments, the dose of Var55 is maintained if the expression level of the biomarker in the second biological sample is higher than the expression level of the biomarker in the first biological sample. In some embodiments, the biomarker is selected from the group consisting of PDGFRβ, pPDGFRβ, MYDGF, pSMAD2 / 3, PROF1, CALM2, CALR, PPIA, IF5A1, soluble RAGE, LGALS1, ANXA5, FABP5, GSTP1, ENO1, YWHAG, LGALS7, IL-11, MMP-2, CXCL7, sCD163, and p-mTOR. In some embodiments, the biomarker is MYDGF. In some embodiments, the biomarker is soluble RAGE. In some embodiments, the biomarker is pSMAD2 / 3. In some embodiments, the biomarker is PDGFRβ. In some embodiments, the interstitial lung disease is idiopathic pulmonary fibrosis.

[0230] Biomarker detection methods

[0246] General methods for determining biomarker expression in biological samples are well known in the art. Methods for detecting biomarkers generally include quantitative methods for determining the level of biomarkers in a biological sample (quantitative methods) or qualitative methods for determining the presence or absence of biomarkers in a biological sample. It is generally known to those skilled in the art which of the following methods is suitable for the qualitative and / or quantitative detection of biomarkers. Biological samples can be assayed for protein biomarkers using, for example, mass spectrometry and immunoassays (Western blotting, ELISA, radioimmunoassay (RIA), fluorescence-based immunoassays, immunoprecipitation assays, flow cytometry, or immunohistochemistry); and for mRNA or DNA biomarkers using Northern blotting, dot blotting, polymerase chain reaction (PCR) analysis, array hybridization (e.g., insight hybridization), RNase protection assays, or DNA SNP chip microarrays. More appropriate methods for detecting biomarkers include measuring physical or chemical properties specific to the biomarker, such as its exact molecular weight or NMR spectrum. Examples of methods for measuring the physical or chemical properties of biomarkers include, but are not limited to, enzyme assays, cytological assays, biosensors, optical devices coupled to immunoassays, biochips, and analytical instruments (such as mass spectrometers, NMR analyzers, and chromatography instruments).

[0231]

[0247] In some embodiments, biomarkers can be amplified by any method commonly known in the art. The term “amplification” refers to the generation of multiple biomarker molecules from a target biomarker. For example, if the biomarker is a nucleic acid, amplifying the nucleic acid refers to the generation of multiple nucleic acid molecules from the target nucleic acid, where primers hybridize to specific sites on the target nucleic acid molecule to provide an initiation site for extension, for example, by polymerase. Amplification can be carried out by any method commonly known in the art, including but not limited to PCR-based amplification methods such as standard PCR, long-chain PCR, hot-start PCR, qPCR, and RT-PCR; isothermal amplification methods such as nucleic acid sequence-based amplification (NASBA) and transcription-mediated amplification (TMA); and hybridization signal amplification methods such as branched DNA assays.

[0232]

[0248] In some embodiments, the biomarkers described herein are detected or measured in biological samples using quantitative real-time PCR (qRT-PCR). qRT-PCR is an amplification technique that can be used to measure levels of mRNA expression (see, e.g., Gibson et al., 1996, Genome Research 6:995-1001, Heid et al., 1996, Genome Research 6:986-994). qRT-PCR assesses the level of PCR product accumulation during amplification. This technique enables quantitative assessment of mRNA levels in multiple samples. For mRNA levels, mRNA is extracted from biological samples, and cDNA is prepared using standard techniques. Primers and fluorescent probes can be designed for the biomarker of interest, and the optimal concentrations of primers and probes can be determined by those skilled in the art. A standard curve can be constructed using the cycle threshold (Ct) value determined by qRT-PCR, which relates to the initial concentration of the target nucleic acid used in the assay. The Ct value is determined by the number of cycles at which exponential amplification of the cDNA molecule begins and is used to determine the expression of the target nucleic acid (i.e., biomarker) in a biological sample.

[0233]

[0249] In some embodiments, RNA or protein samples are normalized for differences in the amount of RNA or protein being assayed, variability in the quality of the RNA or protein samples used, and / or variability between assay runs. In some embodiments, RNA or protein samples are normalized by measuring and incorporating the expression of housekeeping genes (also called internal controls). Examples of housekeeping genes include, but are not limited to, PPIA, ACTB, GAPDH, TFRC, and 18S. In some embodiments, normalization is based on the mean or median signal of all or a large subset of the assayed genes (global normalization approach). For example, the expression levels of a biomarker (e.g., MYDGF) and a housekeeping gene can be measured from biological samples from subjects being treated for idiopathic pulmonary fibrosis using caveolin-1 peptide therapy. The resulting biomarker detection data can be normalized against the housekeeping gene detection data. The mean and standard deviation of the subjects can be calculated. The expression levels of the same biomarker from control samples (e.g., untreated subjects with idiopathic pulmonary fibrosis) can be detected using the same method, and the mean and standard deviation of the control samples can be calculated. The relative expression levels of the biomarker can be compared between subjects treated for idiopathic pulmonary fibrosis and untreated subjects with idiopathic pulmonary fibrosis.

[0234]

[0250] In some embodiments, mRNA biomarkers are detected or measured in biological samples using DNA arrays, chips, or microarrays. Oligonucleotides corresponding to the biomarker's cDNA are immobilized on a chip and then hybridized with labeled nucleic acids derived from a sample obtained from a patient. A sample containing the biomarker transcript yields a positive hybridization signal. For example, to detect or measure an mRNA biomarker, mRNA is extracted from a biological sample, reverse transcribed, and a fluorescently labeled cDNA probe is generated. A microarray capable of hybridizing to the biomarker's cDNA is then probed with the labeled cDNA probe, the slide is scanned, and the fluorescence intensity is measured. This intensity correlates with the hybridization intensity and expression level. Quantitative microarray hybridization assays are well known in the art (see, for example, U.S. Patents 6,004,755 and 6,492,122).

[0235]

[0251] In some embodiments, the biomarkers described herein are detected or measured using mass spectrometry methods such as MALDI / TOF (time-of-flight), SELDI / TOF, liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography-mass spectrometry (HPLC-MS), capillary electrophoresis-mass spectrometry, nuclear magnetic resonance spectrometry, or tandem mass spectrometry (e.g., MS / MS, MS / MS / MS, ESI-MS / MS, etc.). See, for example, U.S. Patent Applications Publications 20030199001; 20030134304; and 20030077616.

[0236]

[0252] In some embodiments, the biomarkers described herein are detected or measured in biological samples using two-dimensional gel electrophoresis (2DE). 2DE separates proteins in biological samples based on molecular charge and molecular weight. Proteins are first separated by charge by isoelectric focusing (IEF) and then further separated by mass by SDS-PAGE. Proteins on the gel separated by IEF are negatively charged by SDS treatment, and the gel is horizontally inserted into an SDS-PAGE gel for electrophoresis. Thus, proteins focused on pI are separated by molecular weight. A system known as "ISO-DALT" allows both IEF and SDS-PAGE to be performed simultaneously. See Buyukkoroglu et al., Omics Technologies and Bioengineering, 2018, pp. 317–351.

[0237]

[0253] Mass spectrometry is well-known in the art and is used to quantify and / or identify protein biomarkers (see, e.g., Li et al., 2000, Tibtech 18:151-160; Rowley et al., 2000, Methods 20:383-397; and Kuster and Mann, 1998, Curr. Opin. Structural Biol. 8:393-400). For additional disclosures regarding mass spectrometers, see, e.g., Principles of Instrumental Analysis, 3rd edition., Skoog, Saunders College Publishing, Philadelphia, 1985; and Kirk-Othmer Encyclopedia of Chemical Technology, 4th sup.th ed. Vol. 15, John Wiley & Sons, New York 1995, pp. 1071-1094.

[0238]

[0254] In some embodiments, a gas-phase ion spectrophotometer is used to analyze the biological sample. In some embodiments, laser desorption / ionization mass spectrometry is used to analyze the biological sample. Modern laser desorption / ionization mass spectrometry ("LDI-MS") can be performed in two main variations: matrix-assisted laser desorption / ionization ("MALDI") mass spectrometry and surface-enhanced laser desorption / ionization ("SELDI"). In MALDI, the analyte is mixed with a matrix-containing solution, and a drop of this liquid is placed on the surface of a substrate. The matrix solution then cocrystallizes with the biomolecules. The substrate is inserted into the mass spectrometer. Laser energy is directed to the substrate surface, where the biomolecules are desorbed and ionized without significant fragmentation. See, for example, U.S. Patent No. 5,118,937 and U.S. Patent No. 5,045,694. In SELDI, the substrate surface is modified to actively participate in the desorption process. In one variation, the surface is derivatized with an adsorbent and / or capture reagent that selectively binds to the protein of interest. In another modification, the surface is derivatized with energy-absorbing molecules that do not desorb upon laser irradiation. In yet another modification, the surface is derivatized with molecules containing photodegradation bonds that bind to the target protein and are cleaved upon laser irradiation. In each of these methods, the derivatizer is generally localized to a specific location on the substrate surface to which the sample is applied. See, for example, U.S. Patent No. 5,719,060 and International Publication No. 98 / 59361. The two methods can be combined, for example, by using a SELDI affinity surface to capture the analyte and adding a matrix-containing liquid to the captured analyte to provide an energy-absorbing material.

[0239]

[0255] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using a protein binder. In some embodiments, the protein binder is a ligand that specifically binds to the biomarker protein. Examples of protein binders include, but are not limited to, synthetic peptides, chemicals, small molecules, and antibodies or fragments thereof.

[0240]

[0256] In some embodiments, the biomarkers described herein are detected or measured in biological samples using immunoassays. The term “immunoassay” refers to any detection method that uses at least one specific antibody for the detection and / or quantification of an antigen (e.g., a biomarker disclosed herein). Examples of immunoassays include, but are not limited to, Western blotting, enzyme-linked immunosorbent assay (ELISA), flow cytometry, radioimmunoassay (RIA), competitive immunoassay, non-competitive immunoassay, fluorescence-polarized immunoassay (FPIA), and enzyme multiplexing immunoassay techniques (EMIT). A wide range of immunoassay techniques have been described previously. See, for example, U.S. Patents 4,016,043; 4,424,279; and 4,018,653.

[0241]

[0257] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using a competitive immunoassay. In a competitive immunoassay, an antigen in the biological sample competes with a labeled antigen to bind to an antibody. The amount of labeled antigen bound to the antibody is then measured. In this method, the reaction is inversely proportional to the concentration of the antigen in the biological sample. This is because a larger reaction means there is less unknown antigen available to compete with the labeled antigen.

[0242]

[0258] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using a non-competitive immunoassay. In a non-competitive immunoassay, also called a “sandwich assay,” an antigen in the biological sample binds to a first antibody site, and then a labeled second antibody binds to the antigen, forming a sandwich. The amount of labeled antibody on that site is then measured. In contrast to competitive immunoassays, the results of a non-competitive immunoassay are directly proportional to the concentration of the antigen.

[0243]

[0259] Immunoassays are generally classified into direct and indirect immunoassays. In direct immunoassays, the binding of an antibody to a target antigen is measured directly. This direct assay uses labeling reagents such as fluorescent tags or enzyme-labeled primary antibodies, which can be visualized without further antibody interaction. In indirect assays, an unconjugated primary antibody binds to the antigen, and then a labeled secondary antibody binds to the primary antibody. The secondary antibody is conjugated with an enzyme label, and a chromogenic substrate or fluorescent substrate is added to visualize the antigen. Signal amplification occurs because several secondary antibodies can react with different epitopes on the primary antibody.

[0244]

[0260] In some embodiments, primary and / or secondary antibodies are labeled with detectable moieties. Numerous labels are commercially available and well known to those skilled in the art. Examples of detectable moieties include, but are not limited to, radioisotopes, colloidal gold particles, fluorescent labels, and enzyme-substrate labels.

[0245]

[0261] In some embodiments, the detectable portion is a radioisotope. Examples of radioisotopes include, but are not limited to, 35S, 14C, 125I, 3H, and 131I. Antibodies can be labeled with radioisotopes using techniques described, for example, in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-Interscience, New York, NY, Pubs. (1991), and radioactivity can be measured using scintillation counts.

[0246]

[0262] In some embodiments, the detectable portion is colloidal gold particles.

[0247]

[0263] In some embodiments, the detectable portion is a fluorescent label. Fluorescent labels include, but are not limited to, rare earth chelates (europium chelate), Texas Red, rhodamine, fluorescein, dansyl, lysamine, umbelliferone, phycocrytherin, phycocyanin, or other commercially available fluorescent dye molecules or derivatives thereof. Fluorescence can be quantified using a fluorophotometer. Fluorescent labels can be conjugated to antibodies using, for example, the techniques disclosed in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-Interscience, New York, NY, Pubs. (1991).

[0248]

[0264] In some embodiments, the detectable portion is an enzyme-substrate label, such as those described in U.S. Patent No. 4,275,149. Enzymes generally catalyze a chemical change in a chromogenic substrate, which can be measured using various techniques. For example, an enzyme may catalyze a change in the substrate's color, which can be measured by spectrophotometry. Alternatively, an enzyme may alter the fluorescence or chemiluminescence of the substrate. Examples of enzyme labels that can be electronically excited by a chemical reaction and then emit measurable light (e.g., using a chemiluminescent meter) or donate energy to a fluorescent acceptor include luciferases (e.g., firefly luciferase and bacterial luciferase as described in U.S. Patent No. 4,737,456), luciferin, 2,3-dihydrophthalazinedione, malate dehydrogenase, urease, peroxidase (e.g., horseradish peroxidase (HRPO)), alkaline phosphatase, β-galactosidase, glucoamylase, lysozyme, sugar oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (e.g., uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and others. Techniques for conjugating enzymes into antibodies are described in O'Sullivan et al., Methods for the Preparation of Enzyme-Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (ed. J. Langone & H. Van Vunakis), Academic press, New York, 73:147-166 (1981).

[0249]

[0265] Numerous enzyme-substrate combinations are available to those skilled in the art. For a general review of enzyme-substrate combinations, see, for example, U.S. Patents 4,275,149 and 4,318,980. Examples of enzyme-substrate combinations include, for example, (a) horseradish peroxidase (HRPO) having hydrogen peroxidase as a substrate (where the hydrogen peroxidase is oxidized with a pigment precursor (e.g., orthophenylenediamine (OPD) or 3,3',5,5'-tetramethylbenzidine hydrochloride (TMB)); (b) alkaline phosphatase (AP) with p-nitrophenyl phosphate as a chromogenic substrate; and (c) μ-D-galactosidase (13-D-Gal) having a chromogenic substrate (e.g., p-nitrophenyl-β-D-galactosidase) or a fluorescent substrate (e.g., 4-methylumbelliferyl-β-D-galactosidase).

[0250]

[0266] In some embodiments, the detection label is indirectly conjugated with the antibody. For example, the antibody can bind to biotin, and any of the labels in the four broad categories above can be conjugated with avidin, which binds to biotin (or vice versa). In another example, the antibody conjugates with a small hapten, and one of the labels above conjugates with an anti-hapten antibody. Thus, these labels conjugate with the antibody indirectly.

[0251]

[0267] In some embodiments, the biomarkers described herein are detected or measured in biological samples using sandwich assays. Examples of sandwich assays include non-competitive single-site and two-site sandwich assays, competitive binding assays, and direct binding assays of labeled antibodies to biomarkers. In a typical forward sandwich assay, an unlabeled antibody is immobilized on a solid substrate, and the biological sample to be tested is brought into contact with the binding molecule. After sufficient time has elapsed to allow the formation of an antibody-antigen complex, a secondary antibody labeled with a reporter molecule that is specific to the antigen and can produce a detectable signal is added and incubated, giving sufficient time for the formation of another antibody-antigen-labeled antibody complex. All unreacted material is washed away, and the presence of the antigen is determined by observing the signal produced by the reporter molecule. The results may be qualitative by simple observation of a visible signal or quantifiable by comparison with a control sample containing a known amount of the biomarker. Variations of the forward sandwich assay include simultaneous assays in which both the biological sample and the labeled antibody are added to the binding antibody at the same time.

[0252]

[0268] Another type of sandwich assay involves immobilizing a biomarker from a biological sample onto a solid support, and then exposing the immobilized biomarker to a specific antibody, which may or may not be labeled with a reporter molecule. Depending on the amount of biomarker and the intensity of the reporter molecule signal, the bound biomarker may be detectable by direct labeling with the antibody. Alternatively, a second labeled antibody specific to the first antibody is exposed to the target-first antibody complex to form a tertiary complex of target-first antibody-second antibody. The complex is detected by a signal emitted by the reporter molecule. As used herein, the term “reporter molecule” refers to a molecule that, by its chemical properties, provides an analytically identifiable signal that enables the detection of an antigen-binding antibody. The reporter molecules most commonly used in this type of assay are enzymes, fluorescent dye molecules, or radionuclide-containing molecules (i.e., radioisotopes) and chemiluminescent molecules.

[0253]

[0269] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using ELISA. An ELISA implementation includes at least one antibody having specificity for a particular biomarker. A known amount of anti-biomarker antibody is immobilized on a solid support either nonspecifically (by adsorption to a surface) or specifically (by capture by another antibody specific to the anti-biomarker antibody in a “sandwich” ELISA). After the antigen is immobilized, a detection antibody is added to form a complex with the antigen. The detection antibody can be covalently bound to an enzyme, or it can be detected by a secondary antibody that binds to the enzyme via bioconjugation. Between each step, the plate is washed, typically with a neutral detergent solution, to remove any unspecifically bound proteins or antibodies. After the final washing step, an enzyme substrate is added to develop the plate, generating a visible signal indicating the amount of antigen in the sample. Older ELISAs utilize chromogenic substrates, while newer assays use much more sensitive fluorescent substrates. Numerous ELISA methods and applications are publicly known in the art, and are further described, for example, in Crowther, “Enzyme-Linked Immunosorbent Assay (ELISA),” in Molecular Biomethods Handbook, Rapley et al. (eds.), pp. 595-617, Humana Press, Inc., Totowa, NJ (1998); Harlow and Lane (eds.), Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1988); Ausubel et al. (eds.), Current Protocols in Molecular Biology, Ch.11, John Wiley & Sons, Inc., New York (1994).

[0254]

[0270] In some embodiments, the biomarkers described herein are detected or measured directly in a biological sample using ELISA. For example, in some embodiments, a biological sample containing a biomarker is exposed to a solid support. The biomarker in the biological sample is immobilized on the solid support and directly detected using an enzyme-conjugated antibody specific to the biomarker. A solution containing a suitable enzyme substrate is then added to the antigen-antibody complex. The substrate reacts with the enzyme linked to the antibody, producing a qualitative visual signal, which is usually further quantified by spectrophotometric analysis and serves as an indicator of the amount of biomarker present in the biological sample.

[0255]

[0271] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using indirect ELISA. For example, in some embodiments, a biological sample containing a biomarker is exposed to and immobilized on a solid support (e.g., a microtiter plate well). The biomarker is detected indirectly by first adding an antibody specific to the biomarker, followed by a detection antibody specific to the antibody that specifically binds to the biomarker. This detection antibody can be bound to an enzyme, and in the final step, a substance is added that the enzyme can convert into some detectable signal. For example, in the case of fluorescent ELISA, when the sample is exposed to light, the antigen / antibody complex fluoresces, allowing the amount of antibody in the sample to be measured.

[0256]

[0272] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using a sandwich ELISA. For example, in some embodiments, a capture antibody specific to the biomarker is immobilized on a solid support (e.g., a microtiter plate), and then a biological sample containing the biomarker is added to the solid support. If the biomarker is present in the biological sample, it binds to the capture antibody present on the solid support. In some embodiments, the sandwich ELISA is a “direct sandwich” ELISA, where the captured biomarker is detected directly by using an enzyme-conjugated antibody against the biomarker. Alternatively, in other embodiments, the sandwich ELISA is an “indirect sandwich” ELISA, where the captured biomarker is detected indirectly by using an antibody against the biomarker, and then by another enzyme-conjugated antibody that binds to the biomarker-specific antibody, thus forming an antibody-biomarker-antibody-antibody complex. A suitable reporter reagent is then added to detect the third antibody. In some embodiments, any number of additional antibodies are added as needed to detect the biomarker-antibody complex. In some embodiments, the additional antibodies are labeled or tagged to allow visualization and / or quantification of the biomarker.

[0257]

[0273] In some embodiments, the biomarkers described herein are detected using an immunohistochemistry assay. Immunohistochemical staining of tissue sections has been shown to be a reliable method for assessing or detecting the presence of a protein in a sample. In immunohistochemistry techniques, antibodies are utilized to localize and visualize cellular antigens in situ, generally by chromogenic or fluorescent methods. Examples of methods used to visualize the antibodies include, for example, enzymes conjugated to the antibodies (e.g., luciferase, alkaline phosphatase, horseradish peroxidase, or β-galactosidase), or chemical methods (e.g., DAB / substrate chromogen). The sample is then analyzed microscopically, most preferably by optical microscopy of the sample stained with a stain that is detectable in the visible spectrum, using any of a variety of such staining methods and reagents known to those skilled in the art.

[0258]

[0274] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using Western blotting. In Western blot techniques, gel electrophoresis is used to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or the 3D structure of the protein (native / non-denaturing conditions). The proteins are then transferred to a membrane (typically nitrocellulose or PVDF), where they are detected using an antibody specific for the target protein. In some embodiments, the biomarker is quantified by densitometry.

[0259]

[0275] In some embodiments, the biomarkers described herein are detected or measured using a multiplex assay. A multiplex assay is a type of immunoassay that uses magnetic beads to simultaneously measure multiple analytes in a single experiment. In some embodiments, a multiplex assay is used to detect or measure phosphorylated proteins and total endogenous proteins, for example, pSMAD2 / 3 and total SMAD2 / 3.

[0260]

[0276] In some embodiments, the biomarkers described herein are detected using a lateral flow immunoassay test (LFIA), also known as an immunochromatographic assay or strip test. An LFIA is a simple device designed to detect the presence or absence of a target antigen in a liquid sample. An LFIA test is a form of immunoassay in which the sample flows along a solid substrate by capillary action. After the biological sample is applied to the test, it comes into contact with a coloring reagent mixed with the sample, passes through the substrate, and comes into contact with a test line or test zone pre-treated with an antibody or antigen. Depending on the antigen present in the biological sample, the coloring reagent may bind to the test line or test zone. An LFIA is essentially an immunoassay adapted to operate along a single axis in test strip or dipstick format. Strip tests are extremely versatile and can be easily modified by those skilled in the art to detect a vast range of antigens from bodily fluid samples such as serum, blood, and urine. LFIA strip tests are easy to use, require minimal training, and can be incorporated as an element of point-of-care (POCT) diagnostics used in the field.

[0261]

[0277] In some embodiments, the biomarkers described herein are immobilized on a solid support for detection in an immunoassay. In some embodiments, antibodies specific to the biomarkers are immobilized on a solid support for detection of the biomarkers in an immunoassay. The solid support is typically glass or polymer, with the most commonly used polymers being cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene. In some embodiments, the solid support is in the form of a microplate, tube, beads, or any other surface suitable for performing the immunoassay.

[0262]

[0278] In some embodiments, the biomarkers described herein are detected or measured in a biological sample using a functional assay or an activity-based assay. For example, if the biomarker is an enzyme, the expression level of the biomarker would be related to the level of enzyme activity in the biological sample.

[0263] kit

[0279] The present invention also provides a kit adapted for determining the expression levels of biomarkers in biological samples of subjects with a disease or disorder, such as interstitial lung disease.

[0264]

[0280] As used herein, the term "kit" refers to a set of assembled components along with appropriate packaging and instructions for use.

[0265]

[0281] In some embodiments, the kit includes containers for the components of the kit. In some embodiments, the kit further includes one or more additional containers containing the components of the kit. In some embodiments, one or more additional containers include materials such as reagents (e.g., chromogenic compounds), buffers (e.g., blocking buffers, washing buffers, and substrate buffers), control samples, calibrators, diluents, filters, needles, and syringes.

[0266]

[0282] In some embodiments, the kit includes a container for holding or storing the biological sample (e.g., a sample container or cartridge). In some embodiments, the kit includes one or more containers or reagents for preparing the biological sample. In some embodiments, the kit includes one or more instruments for obtaining the biological sample, such as a syringe, pipette, forceps, or measuring spoon.

[0267]

[0283] In some embodiments, the kit includes reagents for the isolation and / or pretreatment of biological samples. For example, a whole blood sample obtained from a subject may be treated with a pretreatment reagent before an assay used to detect a biomarker. Examples of pretreatment reagents include, but are not limited to, solvents (e.g., methanol and ethylene glycol), salts, and surfactants.

[0268]

[0284] In some embodiments, the kit includes an immunoassay for determining the expression level of a biomarker in a biological sample. In some embodiments, the kit includes at least one capture reagent for the biomarker and at least one detection reagent for measuring the biomarker. In some embodiments, the kit further includes at least one capture reagent for a reference standard and a detection reagent for measuring the reference standard. In some embodiments, the kit detects the expression level of MYDGF, soluble RAGE, or pSMAD2 / 3. For example, in some embodiments, the kit includes a monoclonal antibody specific to MYDGF. MYDGF is captured from the biological sample using the monoclonal antibody directed at MYDGF, and the presence of the captured MYDGF is further detected using a conjugate antibody.

[0269]

[0285] In some embodiments, the kit includes a solid support. In some embodiments, antibodies specific to the biomarkers described herein are coated onto the solid support. Examples of solid supports include, but are not limited to, microparticles, magnetic particles, beads, test tubes, microtiter plates, cuvettes, membranes, films, filter paper, discs, or chips.

[0270]

[0286] In some embodiments, the kit includes quality control elements. Examples of quality control elements include, but are not limited to, sensitivity panels, calibrators, positive controls, and negative controls. Quality control elements are used to establish the performance characteristics of the assay, the integrity of the kit reagents, and / or the standardization of the assay. Calibrators are used to interpolate the concentration of biomarkers in biological samples. Positive controls are recombinant or biological samples that can be detected by the assay. Negative controls are recombinant or biological samples that cannot be detected by the assay. In some embodiments, one or more of the reagents or components of the kit are lyophilized. In some embodiments, the kit further includes one or more reagents (e.g., buffer or water) for the reconstitution of one or more lyophilized components. [Examples]

[0271] Examples

[0287] The present invention will be described in more detail with reference to the following embodiments. These embodiments are provided for illustrative purposes only and are not intended to limit the invention unless otherwise specified. Therefore, this disclosure should not be construed as being limited in any way to the following embodiments, but rather as encompassing any variations that become apparent as a result of the teachings provided herein.

[0272] Example 1: Identification of biomarkers in CSP-7 treated IPF fibroblasts and epithelial cells

[0288] This example describes the identification of biomarkers associated with CSP-7 treatment in fibroblasts and conditionally reprogrammed cells from subjects with IPF.

[0273] Materials and methods Samples and cell cultures:

[0289] Potential biomarkers for CSP-7 were identified using IPF fibroblasts and epithelial cells. Epithelial cells were derived from IPF lung explants using a modified conditional reprogramming protocol (see Liu et al., Am J Pathol, 2012;180, 599-607; and Suprynowixz et al., PNAS, 2012;109:20035-20040) and expanded in vitro. The epithelial cells were SSEA4+EpCAM+ basal progenitor cells, similar to pathogenic EpCAM+ basal-like cells found inside the honeycomb of IPF (see Chilosi et al., Lab Invest, 2002;82:1335-1345).

[0274] CSP-7 treatment:

[0290] IPF fibroblasts and epithelial cells were treated with a 1% DMSO vehicle, 10 μM control peptide (CP), or 10 μM CSP-7 for 24 hours. Cells were then harvested using a standard protocol, and cell lysates were prepared.

[0275] Protein concentration:

[0291] Protein concentrations in PCLS and epithelial cell samples were determined using a BCA assay. Briefly, equal volumes of protein extracts were labeled with fluorescent dyes (matched in size and charge). Spectroscopically degradable dyes allowed for simultaneous co-separation and analysis of protein concentrations on a single multi-layer gel.

[0276] 2D gel electrophoresis and mass spectrometry:

[0292] The 2D gel was prepared using isoelectric focusing (IEF) for the first dimension, and SDS-polyacrylamide gel electrophoresis (SDS-PAGE) (2D-DIGE) for the second dimension. After electrophoresis, the gel was scanned using a Typhoon image scanner. Image presentation data including single images and overlay images was generated using ImageQuant software. Comparative analysis of all spots was performed using DeCyder 'in-gel' or 'cross-gel' analysis software. DMSO vehicle samples were pooled and used as reference samples. The protein expression ratio between CSP-7 or CP and the DMSO vehicle was determined, and p-values were assigned. Spots that were significantly regulated across all samples (P<0.05) were assigned numbers and automatically picked from the 2D gel using an Ettan Spot Picker. The spots were then identified using MALDI-TOF.

[0277]

[0293] ELISA: Commercially available human MYDGF ELISA was used to quantify MYDGF protein expression in the supernatants of CSP-7-treated IPF fibroblasts and epithelial cells, as well as in human plasma.

[0278] Results:

[0294] 37 spots were significantly upregulated in CSP-7-treated IPF fibroblasts, and 27 spots were significantly upregulated in CSP-7-treated epithelial cells. The significantly regulated proteins that were upregulated in both CSP-7-treated IPF fibroblasts and epithelial cells are shown in Table 5 below.

[0279] [Table 7]

[0280]

[0295] Other proteins that were significantly regulated in IPF fibroblasts treated with CSP-7 include galectin-1 (LGALS1), alpha-enolase (EnoA), and annexin V (ANXA5).

[0281]

[0296] Other proteins significantly regulated in epithelial cells treated with CSP-7 include fatty acid-binding protein (FABP5), glutathione S-transferase P (GSTP-1), and 14-3-3 protein gamma.

[0282]

[0297] Additional experiments were conducted to validate the MYDGF biomarkers identified by MALDI-TOF. Epithelial cells derived from IPF lung explants were treated with 1% DMSO vehicle, CSP-7 (10 μM), Var55 (10 μM), control peptide 1 (CP1, 10 μM), control peptide 2 (CP2, 10 μM), pirfenidone (500 μM), or nintedanib (100 nM). MYDGF expression in the epithelial cell supernatant was measured by ELISA. As shown in Figures 1A and 1B, supernatant from epithelial cells treated with 10 μM or 100 μM CSP-7 showed an increase in MYDGF protein.

[0283]

[0298] Overall, these experiments identified numerous biomarkers induced in IPF fibroblasts and epithelial cells in response to CSP-7. Additional experiments confirmed increased MYDGF biomarkers in CSP-7-treated epithelial cells derived from patients with IPF. No changes in MYDGF protein expression were observed in healthy controls treated with CSP-7.

[0284] Example 2. Soluble RAGE as a biomarker for CSP-7 treatment

[0299] The endogenous secretion receptor for advanced glycation end products (esRAGE) is a soluble isoform produced by alternative splicing of the RAGE gene and is reduced in the lung tissue, serum, and BAL of IPF patients with idiopathic pulmonary fibrosis (see Yamaguchi et al., Respiratory Research, 2020; 21:145).

[0285]

[0300] Precision-cut lung sections (PCLS) were used to determine whether soluble RAGE was affected by CSP-7 treatment. PCLS allows for ex vivo testing of therapeutic agents for human IPF lung tissue, such as CSP-7 (Figure 2). Briefly, fresh lungs were obtained from human subjects with IPF, the lung cores were perforated, and the sections were transferred to a Krumdieck slicer. The precision-cut lung sections (PCLS) were kept alive for up to one month with human serum and culture medium.

[0286]

[0301] IPF PCLS were either untreated (UT) or treated with CSP-7 (10 μM or 100 μM), a control peptide (CP, 100 μM), TGFβ (2 ng / mL), or pirfenidone (Pirf, 500 μM) and nintedanib (Nin, 100 nM). Soluble RAGE expression was measured in the supernatant of IPF PCLS on days 2 and 4 after treatment with ELISA (R&D Systems, Cat No. DRG00). ELISA detects the extracellular domain of human RAGE. As shown in Figures 3A and 3B, IPF PCLS treated with 10 μM or 100 μM CSP-7 showed increased soluble RAGE compared to untreated or CP-treated controls. Figure 3C shows the dose-dependent increase in soluble RAGE expression in IPF PCLS on days 2 and 4 after CSP-7 treatment.

[0287]

[0302] LDH activity was also examined in the supernatant of IPF PCLS that were either untreated (UT) or treated with CSP-7 (10 μM or 100 μM), control peptide (CP, 100 μM), TGFβ (2 ng / mL), or pirfenidone (Pirf, 500 μM) and nintedanib (Nin, 100 nM). LDH activity was measured using an LDH assay (Sigma-Aldrich, catalog no. MAK066). Overall, LDH activity was slightly reduced in PCLS treated with CSP-7 compared to the untreated control (Figures 4A and 4B).

[0288]

[0303] Soluble RAGE expression was also examined in healthy subjects on days 1 and 13 after inhalation of the CSP-7 peptide. As shown in Figure 5, soluble RAGE expression in the plasma of healthy subjects treated with CSP-7 increased on days 1 and 13 after CSP-7 treatment.

[0289]

[0304] Overall, these data demonstrate increased soluble RAGE expression in end-stage IPF PCLS cultures after CSP-7 treatment. The reduced LDH activity in CSP-7-PCLS may also indicate healthier explant tissue and increased epithelial cell survival, which is important for epithelial repair and reduced fibrosis.

[0290] Example 3. PDGFRβ as a biomarker for CSP-7 treatment.

[0305] Platelet-derived growth factor (PDGF) signaling has been linked to the pathogenesis of pulmonary fibrosis. Many fibrillation mediators, including TGF-β, IL-1, TNF-α, bFGF, and thrombin, exhibit PDGF-dependent pro-fibrotic activity. Inhibition of PDGF signaling has also been shown to improve bleomycin-induced pulmonary fibrosis in mice. See Abdollahi et al., JEM, 2005 201(6):925-935; and Kishi et al., PLoS One, 2018, 13(12):e0209786.

[0291]

[0306] Experiments were conducted to determine whether PDGFRβ could be used as a biomarker for CSP-7. PCLS were obtained from IPF subjects, cultured as described above, and shown in Figure 2. PCLS were either untreated (UT) or treated with CSP-7 (10 μM or 100 μM), a control peptide (CP, 100 μM), TGFβ (2 ng / mL), or pirfenidone (Pirf, 500 μM) and nintedanib (Nin, 100 nM). PDGFRβ expression was measured in the supernatant of IPF PCLS on days 2 and 4 after treatment by ELISA (R&D Systems, Cat No. DRG00). As shown in Figures 6A and 6B, PCLS treated with 10 μM or 100 μM CSP-7 demonstrated reduced PDGFRβ protein expression compared to the untreated control.

[0292]

[0307] Overall, these data demonstrate a decrease in PDGFRβ expression in end-stage IPF PCLS cultures after CSP-7 treatment. PDGFRβ may be a useful biomarker associated with CSP-7 therapy in patients with fibrosis.

[0293] Example 4. Phosphorylated SMAD2 / 3 as a biomarker for CSP-7 and Var55 treatment.

[0308] TGF-β plays a central role in fibrosis, contributing to the influx and activation of inflammatory cells, the transition of cells from epithelial to mesenchymal (EMT), the influx of fibroblasts, and the subsequent generation of the extracellular matrix. TGF-β transmits signals via SMAD proteins such as SMAD2 / 3. Experiments were conducted to determine whether the phosphorylation status of SMAD2 / 3 can be used as a biomarker associated with CSP-7 and / or Var55 treatment.

[0294]

[0309] Primary pulmonary fibroblasts from IPF patients were treated with 1% DMSO vehicle, CSP-7 (10 μM), Var55 (10 μM), pirfenidone (500 μM), or nintedanib (100 nM). SMAD2 / 3 phosphorylation was measured in IPF fibroblasts by multiplex assay or Western blot analysis, and phosphorylated SMAD2 / 3 was normalized relative to total SMAD2 / 3 protein. As shown in Figure 7A, IPF fibroblasts treated with 10 μM CSP-7 showed reduced SMAD2 / 3 phosphorylation compared to 1% DMSO vehicle or DMEM controls. IPF fibroblasts treated with 10 μM Var55 showed no difference in SMAD2 / 3 phosphorylation.

[0295]

[0310] Additional experiments were conducted to determine whether SMAD2 / 3 phosphorylation could be used as a biomarker associated with Var55 treatment. Epithelial cells from IPF patients were treated with DMEM (control), Var55 (100 μM), nintedanib (100 nM), control peptide 1 (CP1, 100 μM), or control peptide 2 (CP2, 100 μM). SMAD2 / 3 phosphorylation was measured in IPF fibroblasts by multiplex assay or Western blot analysis, and phosphorylated SMAD2 / 3 was normalized to total SMAD2 / 3 protein. As shown in Figure 7B, IPF epithelial cells treated with 100 μM Var55 showed decreased SMAD2 / 3 phosphorylation compared to the DMEM control.

[0296]

[0311] In summary, these data demonstrate a decrease in SMAD2 / 3 phosphorylation in IPF fibroblasts and epithelial cells after treatment with CSP-7 and Var55. Therefore, SMAD2 / 3 phosphorylation may be a useful biomarker associated with CSP-7 and / or Var55 treatment in patients with fibrosis.

[0297] Example 5. Galectin-7 as a biomarker for CSP-7 and Var55 treatment.

[0312] Basal-like cells and fibroblasts derived from either normal healthy subjects or subjects with idiopathic pulmonary fibrosis (IPF) were treated with DMSO vehicle, CSP-7 (10 μM), Var55 (10 μM), control peptide 1 (CP1, 10 μM), control peptide 2 (CP2, 10 μM), pirfenidone (500 μM), or nintedanib (100 nM). Galectin-7 was measured by ELISA. The results of the study are shown in Figure 8.

[0298] Example 6. Biomarker of modified Cav-1 peptide in an aged mouse model of pulmonary fibrosis

[0313] The objective of this study was to evaluate the modified Cav-1 peptide CSP-7 in bleomycin models of idiopathic pulmonary fibrosis and acute respiratory distress syndrome (ARDS). Aged male mice were selected that closely matched the human population most vulnerable to ARDS.

[0299]

[0314] Bleomycin (BLM, 8 U / kg) was intratracheally administered to 73-74 week old mice (n=52), predicting a 50% reduction in mice due to BLM-induced lung injury. On day 14 after infusion, CSP-7 and a control peptide (CP) were administered to the mice by dry powder inhalation (DPI) or intraperitoneal injection (IP), as shown in Table 6 below. The mice were sacrificed on day 21 after infusion, and blood and tissue samples were collected for analysis. Statistical analysis was performed using Dunnett's multiple comparison test.

[0300] [Table 8]

[0301]

[0315] Figure 9 shows collagen in whole lung homogenates of aged mice treated with physiological saline or BLM after being administered with a control peptide (CP) or CSP-7 by dry powder inhalation (DPI). BLM-treated mice administered with CSP-7 showed a decrease in total collagen in the lungs compared to mice administered with the control peptide.

[0302]

[0316] Figure 10A shows the total SMAD (tSMAD, upper panel) and phosphorylated SMAD (pSMAD, lower panel) in whole lung homogenates of aged mice treated with BLM, administered by dry powder inhalation (DPI) with a control peptide (CP) or CSP-7, or by intraperitoneal injection (IP) with CSP-7.

[0303]

[0317] Figure 10B shows galectin-7 in whole lung homogenates of BLM-treated aged mice that were administered either control peptide (CP) or CSP-7 by dry powder inhalation (DPI), or CSP-7 by intraperitoneal injection (IP).

[0304] Example 7. CSP-7 attenuates receptor tyrosine kinase signaling in IPF.

[0318] The objective of this study was to investigate the effect of CSP-7 treatment on receptor tyrosine kinase signaling in IPF fibroblasts.

[0305]

[0319] End-stage IPF fibroblasts and non-cancerous donor lung fibroblasts were obtained and cultured in vitro. Initial passages from normal donor and IPF fibroblasts were either left untreated or treated with DMSO, a control peptide, or 10 μM CSP-7 for 24 hours. Cells were then harvested for a reverse-phase protein array. This array contains 240 validated antibodies against total protein and phosphoproteins, epithelial-mesenchymal transition (EMT), stem cells, apoptosis, DNA damage, autophagy, proliferation and cell cycle, growth factor receptors, cytokines / STAT, nuclear receptor / transcriptional regulatory proteins, and metabolome enzymes. Nine technical replicates were performed for each sample, and the signals were normalized relative to total protein.

[0306]

[0320] Figure 11A shows that IPF treated with CSP-7 showed decreased phosphorylation of EGFR, SRC, MEK1 / 2, and Raptor, while CSP-7 treatment of normal lung fibroblasts did not affect these signaling pathways.

[0307]

[0321] Figure 11B shows that IPF treated with CSP-7 showed suppression of p-PDGFRβ and p-mTOR compared to untreated IPF or IPF treated with DMSO or a control peptide.

[0308]

[0322] Figure 11C provides an overview of the reverse-phase protein array used to evaluate receptor tyrosine kinase (RTK) signaling, metabolic signaling, infiltration-related markers, and histone deacetylase (HDAC).

[0309]

[0323] Overall, these results indicate that CSP-7 treatment of IPF reduces receptor tyrosine kinase signaling, metabolic signaling, infiltration-related markers, and histone deacetylase (HDAC) activation. Of particular interest is the suppression of p-PDGFRβ as a p-mTOR signaling pathway, as shown in Figure 11B, which is known to contribute to myofibroblast support. These results suggest that while CSP-7 acts as a sponge of abnormal signaling when administered to IPF, it may have limited effects on fibroblasts with normal signaling.

[0310] Example 8. IL-11 as a biomarker for modified Cav-1 peptide therapy

[0324] The objective of this study was to evaluate the effect of CSP-7 treatment on IL-11 expression in the lungs of mice with acute lung injury.

[0311]

[0325] Mice were treated with bleomycin to induce acute lung injury. Mice treated with saline alone were used as negative controls. Mice were assigned to either a prophylactic treatment group or a therapeutic treatment group. The prophylactic treatment group received a control peptide or CSP-7 three times daily before bleomycin administration. The therapeutic group received a control peptide or CSP-7 three times daily after bleomycin administration. Mice were sacrificed four days after bleomycin treatment, and bronchoalveolar lavage fluid (BALF) was collected from the mice for IL-11 measurement.

[0312]

[0326] As shown in Figure 12A, bleomycin-treated mice in the prophylactic treatment group administered with CSP-7 showed reduced IL-11 levels in BALF compared to bleomycin-treated mice administered with a control peptide. As shown in Figure 12B, bleomycin-treated mice in the therapeutic treatment group administered with CSP-7 showed reduced IL-11 levels in BALF compared to bleomycin-treated mice administered with a control peptide.

[0313] Example 9. sCD163 and MMP-2 as biomarkers for modified Cav-1 peptide therapy

[0327] The objective of this study was to evaluate the effects of CSP-7 treatment on sCD163 and MMP-2 expression in lung macrophages from individuals with IPF.

[0314]

[0328] Lung macrophages were collected from individuals with IPF and cultured in vitro. Lung macrophages were either left untreated or treated with 0.1 μM or 1 μM CSP-7. One day after treatment, the supernatant from the lung macrophage cultures was collected and analyzed for sCD163 and MMP-2.

[0315]

[0329] As shown in Figure 13A, IPF macrophages treated with CSP-7 showed a decrease in sCD163 in the supernatant compared to untreated controls. As shown in Figure 13B, IPF macrophages treated with CSP-7 showed a decrease in MMP-2 in the supernatant compared to untreated controls.

[0316] Example 10. CXCL7 as a biomarker for modified Cav-1 peptide therapy

[0330] The objective of this study was to evaluate the effect of Var55 treatment on CXCL7 expression in lung macrophages from individuals with IPF.

[0317]

[0331] Lung macrophages were collected from humans with IPF or healthy controls and cultured in vitro. Lung macrophages were either left untreated or treated with Var55. After treatment, supernatant was collected from the lung macrophage cultures and analyzed for CXCL7 expression by ELISA. As shown in Figure 14, healthy controls (triangles) and IPF macrophages (circles) treated with Var55 showed a significant decrease in CXCL7 in the supernatant.

[0318] Reference All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entirety for any purpose. However, no reference to any references, articles, publications, patents, patent publications, or patent applications cited herein constitutes, and should not be construed as, an endorsement or any suggestion that they constitute valid prior art or form part of common technical knowledge in any country of the world.

Claims

1. A method for treating a subject with caveolin-1 peptide therapy, wherein the subject suffers from fibrosis, and the method is (a) Obtaining a biological sample from the subject, or the step of obtaining one; (b) Treating the cells of the biological sample with caveolin-1 peptide or a derivative thereof, or a step in which the cells have been treated; (c) the step of measuring the expression level of the biomarker in the cells; and (d) A step of comparing the expression level of the biomarker with that of a control sample. The biomarker is one of the following: myelo-derived growth factor (MYDGF), soluble RAGE, phosphorylated mothers-against decapentaplesic homolog 2 / 3 (pSMAD2 / 3), platelet-derived growth factor receptor beta (PDGFRβ), galectin-7 (LGALS7), interleukin-11 (IL-11), matrix metalloproteinase-2 (MMP-2), chemokine ligand 7 (CXCL-7), soluble CD163, phosphorylated mTOR (p-mTOR), phosphorylated PDGFRβ (pPDGFRβ), prolifen (PROF1), calmodulin 2 (CALM2), calreticulin (CALR), peptidyl-prolyl cis-trans isomerase A (PPIA), or eukaryotic translation initiation factor 5A (EIF5A1); If the expression levels of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 increase, administer caveolin-1 peptide or a derivative thereof to the subject; or A method comprising administering a caveolin-1 peptide or a derivative thereof to a subject when the expression level of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, soluble CD163, or pSMAD2 / 3 decreases.

2. The method according to claim 1, wherein the control sample is a cell obtained from the subject before treatment with the caveolin-1 peptide or a derivative thereof.

3. The method according to claim 1, wherein the control sample is cells obtained from a subject or population of subjects having interstitial lung disease.

4. The method according to claim 1, wherein the control sample is cells obtained from a healthy subject.

5. The method according to any one of claims 1 to 4, wherein the cells are fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal-like cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphocytes, T cells, monocytes, NK cells, dendritic cells, or peripheral blood mononuclear cells (PBMCs).

6. The method according to claim 5, wherein the cell is a fibroblast.

7. The method according to claim 5, wherein the cells are basal-like cells.

8. The method according to any one of claims 1 to 7, wherein the cells are treated with the caveolin-1 peptide or a derivative thereof, ex vivo or in vitro.

9. The method according to any one of claims 1 to 8, wherein the caveolin-1 peptide or its derivative is FTTFTVT (SEQ ID NO: 3).

10. The method according to any one of claims 1 to 9, wherein the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTTVTKGSaaNH2 (SEQ ID NO: 4).

11. The method according to any one of claims 1 to 10, wherein the fibrosis is interstitial lung disease, hepatic fibrosis, renal fibrosis, dermatofibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataracts, retina and vitreoretinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloids and hypertrophic scars, peritoneal fibrous disease, chronic obstructive pulmonary disease, postoperative fibroids, diabetic nephropathy, gynecological cancer, myeloproliferative syndrome, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome.

12. The method according to claim 11, wherein the interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar injury, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease.

13. The method according to claim 12, wherein the interstitial lung disease is idiopathic pulmonary fibrosis.

14. The method according to any one of claims 1 to 13, wherein the expression level of the biomarker is measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

15. The method according to any one of claims 1 to 14, wherein if the expression level of MYDGF increases compared to the control sample, the subject is administered caveolin-1 peptide or a derivative thereof.

16. The method according to claim 15, wherein the expression level of MYDGF increases by approximately 1.5 times, approximately 2 times, approximately 3 times, approximately 4 times, approximately 5 times, or more compared to the control sample.

17. The method according to any one of claims 1 to 14, wherein if the expression level of soluble RAGE increases compared to the control sample, the subject is administered caveolin-1 peptide or a derivative thereof.

18. The method according to claim 17, wherein the expression level of soluble RAGE increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the control sample.

19. The method according to any one of claims 1 to 14, wherein if the expression level of pSMAD2 / 3 decreases compared to the control sample, the subject is administered caveolin-1 peptide or a derivative thereof.

20. The method according to claim 19, wherein the expression level of pSMAD2 / 3 is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

21. The method according to any one of claims 1 to 14, wherein if the expression level of PDGFRβ and / or pPDGFRβ decreases compared to the control sample, the subject is administered a caveolin-1 peptide or a derivative thereof.

22. The method according to claim 21, wherein the expression level of PDGFRβ and / or pPDGFRβ is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

23. The method according to any one of claims 1 to 22, further comprising measuring the expression level of an internal control, wherein the expression level of the internal control is not affected by the caveolin-1 peptide or its derivative.

24. The method according to any one of claims 1 to 23, wherein steps (b) to (d) of the above method are repeated once or more times.

25. A method for identifying changes in the expression levels of biomarkers associated with caveolin-1 peptide therapy, wherein the method is (a) Treating target cells with interstitial lung disease with caveolin-1 peptide or its derivatives; (b) Measuring the expression level of the biomarker in the cells; and (c) Comparing the expression level of the biomarker with that of a control sample. The caveolin-1 peptide or its derivative modulates the expression level of the biomarker in the cells; A method wherein the biomarker is bone marrow-derived growth factor (MYDGF), soluble RAGE, phosphorylated mothers-against decapentaplesic 2 / 3 homolog (pSMAD2 / 3), platelet-derived growth factor receptor beta (PDGFRβ), galectin-7 (LGALS7), interleukin-11 (IL-11), matrix metalloproteinase-2 (MMP-2), chemokine ligand 7 (CXCL-7), soluble CD163, phosphorylated mTOR (p-mTOR), phosphorylated PDGFRβ (pPDGFRβ), prolifen (PROF1), calmodulin 2 (CALM2), calreticulin (CALR), peptidyl-prolyl cis-trans isomerase A (PPIA), or eukaryotic translation initiation factor 5A (EIF5A1).

26. The method according to claim 25, wherein the control sample is cells obtained from the subject having interstitial lung disease prior to treatment with the caveolin-1 peptide or its derivative.

27. The method according to claim 25, wherein the control sample is cells obtained from a subject or population of subjects having interstitial lung disease.

28. The method according to claim 25, wherein the control sample is cells obtained from a healthy subject.

29. The method according to any one of claims 25 to 28, wherein the cells are fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal-like cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphocytes, T cells, monocytes, NK cells, dendritic cells, or peripheral blood mononuclear cells (PBMCs).

30. The method according to claim 29, wherein the cell is a fibroblast.

31. The method according to claim 29, wherein the cells are basal-like cells.

32. The method according to any one of claims 25 to 31, wherein the cells are treated with the caveolin-1 peptide or a derivative thereof ex vivo or in vitro.

33. The method according to claim 32, wherein the cells are obtained from the subject having interstitial lung disease.

34. The method according to any one of claims 25 to 33, wherein the caveolin-1 peptide or its derivative is FTTFTVT (SEQ ID NO: 3).

35. The method according to any one of claims 25 to 33, wherein the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTTVTKGSaaNH2 (SEQ ID NO: 4).

36. The method according to any one of claims 25 to 35, wherein the subject is a human.

37. The method according to any one of claims 25 to 36, wherein the interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar injury, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease.

38. The method according to claim 37, wherein the interstitial lung disease is idiopathic pulmonary fibrosis.

39. The method according to any one of claims 25 to 38, wherein the expression level of the biomarker is measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

40. The method according to any one of claims 25 to 39, wherein the increase in the expression level of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 is associated with caveolin-1 therapy.

41. The method according to any one of claims 25 to 40, wherein the reduction in the expression levels of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 is associated with caveolin-1 therapy.

42. The method according to any one of claims 25 to 40, wherein the expression level of MYDGF is increased compared to the control sample.

43. The method according to claim 42, wherein the expression level of MYDGF increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the control sample.

44. The method according to any one of claims 25 to 40, wherein the expression level of soluble RAGE is increased compared to the control sample.

45. The method according to claim 44, wherein the expression level of soluble RAGE increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the control sample.

46. The method according to any one of claims 25 to 40, wherein the expression level of pSMAD2 / 3 is reduced compared to the control sample.

47. The method according to claim 46, wherein the expression level of pSMAD2 / 3 is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

48. The method according to any one of claims 25 to 40, wherein the expression level of PDGFRβ and / or pPDGFRβ is reduced compared to the control sample.

49. The method according to claim 48, wherein the expression level of PDGFRβ and / or pPDGFRβ is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

50. The method according to any one of claims 25 to 49, further comprising measuring the expression level of an internal control, wherein the expression level of the internal control is not affected by the caveolin-1 peptide or its derivative.

51. The method according to any one of claims 25 to 50, wherein steps (a) to (c) of the above method are repeated once or more times.

52. A method for predicting or determining the effectiveness of a therapeutic agent, (a) Treating cells of a subject with interstitial lung disease with the therapeutic agent; (b) Measuring the expression level of the biomarker in the cells of the subject; The biomarker is MYDGF, soluble RAGE, pSMAD2 / 3, PDGFRβ, pPDGFRβ, LGALS7, IL-11, MMP-2, CXCL7, sCD163, p-mTOR, PROF1, CALM2, CALR, PPIA, or EIF5A1; and (c) Comparing the expression level of the biomarker with that of a control sample. Methods that include...

53. The method according to claim 52, wherein the control sample is cells obtained from the subject having interstitial lung disease before treatment with the therapeutic activator.

54. The method according to claim 52, wherein the control sample is cells obtained from a subject or population of subjects having interstitial lung disease.

55. The method according to claim 52, wherein the control sample is cells obtained from a healthy subject.

56. The method according to any one of claims 52 to 55, wherein the cells are fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal-like cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphocytes, T cells, monocytes, NK cells, dendritic cells, or PBMCs.

57. The method according to claim 56, wherein the cell is a fibroblast.

58. The method according to claim 56, wherein the cell is a basal-like cell.

59. The method according to any one of claims 52 to 58, wherein the cells are treated with the therapeutic activator ex vivo or in vitro.

60. The method according to claim 59, wherein the cells are obtained from the subject having interstitial lung disease.

61. The method according to any one of claims 52 to 60, wherein the therapeutic activator is a small molecule.

62. The method according to any one of claims 52 to 60, wherein the therapeutic activator is a biological agent.

63. The method according to claim 62, wherein the biological agent is caveolin-1 peptide or a derivative thereof.

64. The method according to claim 63, wherein the caveolin-1 peptide or its derivative is FTTFTVT (SEQ ID NO: 3).

65. The method according to claim 63, wherein the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTTVTKGSaaNH2 (SEQ ID NO: 4).

66. The method according to any one of claims 52 to 65, wherein the subject is a human.

67. The method according to any one of claims 52 to 66, wherein the interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar injury, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease.

68. The method according to claim 67, wherein the interstitial lung disease is idiopathic pulmonary fibrosis.

69. The method according to any one of claims 52 to 68, wherein the expression level of the biomarker is measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

70. The method according to any one of claims 52 to 69, wherein an increase in the expression level of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 indicates a favorable response to the therapeutic activator.

71. The method according to any one of claims 52 to 69, wherein a decrease in the expression level of PDGFRβ, pPDGFRβ, p-mTOR, pSMAD2 / 3, IL-11, MMP-2, CXCL7, sCD163, or CALR indicates a favorable response to the therapeutic activator.

72. The method according to any one of claims 52 to 69, wherein the expression level of MYDGF is increased compared to the control sample.

73. The method according to claim 72, wherein the expression level of MYDGF increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the control sample.

74. The method according to any one of claims 52 to 69, wherein the expression level of soluble RAGE is increased compared to the control sample.

75. The method according to claim 74, wherein the expression level of soluble RAGE increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the control sample.

76. The method according to any one of claims 52 to 69, wherein the expression level of pSMAD2 / 3 is reduced compared to the control sample.

77. The method according to claim 76, wherein the expression level of pSMAD2 / 3 is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

78. The method according to any one of claims 52 to 69, wherein the expression level of PDGFRβ and / or pPDGFRβ is reduced compared to the control sample.

79. The method according to claim 78, wherein the expression level of PDGFRβ and / or pPDGFRβ is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

80. The method according to any one of claims 52 to 79, further comprising measuring the expression level of an internal control, wherein the expression level of the internal control is not affected by the therapeutic activator.

81. The method according to any one of claims 52 to 80, wherein steps (a) to (c) of the above method are repeated once or more times.

82. A method for predicting or determining the efficacy of caveolin-1 peptide or its derivatives, (a) Treating biological samples from subjects with fibrosis with caveolin-1 peptide or its derivatives; (b) Measuring the expression level of the biomarker in the biological sample from the subject; The biomarker is MYDGF, soluble RAGE, pSMAD2 / 3, PDGFRβ, pPDGFRβ, LGALS7, IL-11, MMP-2, CXCL7, sCD163, p-mTOR, PROF1, CALM2, CALR, PPIA, or IF5A1; and (c) Comparing the expression level of the biomarker with that of a control sample. Methods that include...

83. The method according to claim 82, wherein the control sample is a biological sample obtained from the subject having fibrosis before treatment with the caveolin-1 peptide or its derivative.

84. The method according to claim 82, wherein the control sample is a biological sample obtained from a subject or population of subjects having fibrosis.

85. The method according to claim 82, wherein the control sample is a biological sample obtained from a healthy subject.

86. The method according to any one of claims 82 to 85, wherein the biological sample is bronchoalveolar lavage fluid (BALF), lung tissue, fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphocytes, T cells, monocytes, NK cells, dendritic cells, or PBMCs.

87. The method according to claim 86, wherein the biological sample is a fibroblast.

88. The method according to claim 86, wherein the biological sample is a basal-like cell.

89. The method according to claim 86, wherein the biological sample is lung tissue.

90. The method according to any one of claims 82 to 85, wherein a biological sample is treated with the caveolin-1 peptide or a derivative thereof, ex vivo or in vitro.

91. The method according to claim 90, wherein the biological sample is obtained from the subject having fibrosis.

92. The method according to any one of claims 82 to 91, wherein the caveolin-1 peptide or its derivative is FTTFTVT (SEQ ID NO: 3).

93. The method according to any one of claims 82 to 91, wherein the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTTVTKGSaaNH2 (SEQ ID NO: 4).

94. The method according to any one of claims 82 to 93, wherein the subject is a human.

95. The method according to any one of claims 82 to 94, wherein the fibrosis is interstitial lung disease, hepatic fibrosis, renal fibrosis, dermatofibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataracts, retina and vitreoretinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloids and hypertrophic scars, peritoneal fibrous disease, chronic obstructive pulmonary disease, postoperative fibroids, diabetic nephropathy, gynecological cancer, myeloproliferative syndrome, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome.

96. The method according to claim 95, wherein the interstitial lung disease is idiopathic pulmonary fibrosis, lymphangioleiomyomatosis, nonspecific interstitial pneumonia, idiopathic interstitial pneumonia, idiopathic organizing pneumonia, acute interstitial lung disease, respiratory bronchiolitis-associated interstitial lung disease, desquamative interstitial pneumonia, lymphocytic interstitial pneumonia, pulmonary sarcoidosis, diffuse alveolar injury, systemic sclerosis, polymyositis, systemic lupus erythematosus, rheumatoid arthritis, drug-induced interstitial lung disease, or occupational interstitial lung disease.

97. The method according to claim 96, wherein the interstitial lung disease is idiopathic pulmonary fibrosis.

98. The method according to any one of claims 82 to 97, wherein the expression level of the biomarker is measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

99. The method according to any one of claims 82 to 98, wherein an increase in the expression level of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, soluble RAGE, or EIF5A1 shows a favorable reaction to the caveolin-1 peptide or its derivative.

100. The method according to any one of claims 82 to 98, wherein a decrease in the expression level of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicates a favorable response to the caveolin-1 peptide or its derivative.

101. The method according to any one of claims 82 to 98, wherein the expression level of MYDGF is increased compared to the control sample.

102. The method according to claim 101, wherein the expression level of MYDGF increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the control sample.

103. The method according to any one of claims 82 to 98, wherein the expression level of soluble RAGE is increased compared to the control sample.

104. The method according to claim 103, wherein the expression level of soluble RAGE increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the control sample.

105. The method according to any one of claims 82 to 98, wherein the expression level of pSMAD2 / 3 is reduced compared to the control sample.

106. The method according to claim 105, wherein the expression level of pSMAD2 / 3 is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

107. The method according to any one of claims 82 to 98, wherein the expression level of PDGFRβ and / or pPDGFRβ is reduced compared to the control sample.

108. The method according to claim 107, wherein the expression level of PDGFRβ and / or pPDGFRβ is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

109. The method according to any one of claims 82 to 108, further comprising measuring the expression level of an internal control, wherein the expression level of the internal control is not affected by the caveolin-1 peptide or its derivative.

110. The method according to any one of claims 82 to 109, wherein steps (a) to (c) of the method are repeated once or more times.

111. A method for predicting or determining the efficacy of caveolin-1 peptide or its derivatives in subjects with fibrosis, (a) Obtaining a first biological sample from the subject before treatment with the caveolin-1 peptide or its derivative; (b) Administering the caveolin-1 peptide or its derivative to the subject; (c) Obtaining a second biological sample from the subject after treatment with the caveolin-1 peptide or its derivative; (d) Measuring the expression levels of biomarkers in the first biological sample and the second biological sample; The biomarker is MYDGF, soluble RAGE, pSMAD2 / 3, PDGFRβ, pPDGFRβ, LGALS7, IL-11, MMP-2, CXCL7, sCD163, p-mTOR, PROF1, CALM2, CALR, PPIA, or IF5A1; and (e) Comparing the expression level of the biomarker in the first biological sample with the expression level of the biomarker in the second biological sample. Methods that include...

112. The method according to claim 111, wherein step (a) of the method further comprises determining the expression level of the biomarker in the first biological sample by comparison with a control sample before treatment with the caveolin-1 peptide or a derivative thereof.

113. The method according to claim 112, wherein the control sample is a biological sample obtained from a subject or population of subjects having fibrosis.

114. The method according to claim 112 or 113, wherein the expression level of the biomarker in the first biological sample of step (a) is used to determine the optimal dose of the caveolin-1 peptide or its derivative administered in step (b).

115. The method according to any one of claims 111 to 114, wherein the biological sample is serum, plasma, BALF, lung tissue, fibroblasts, type I alveolar epithelial cells, type II alveolar epithelial cells, basal cells, Clara cells, ciliated cells, club cells, goblet cells, neuroendocrine cells, endothelial cells, bialveolar stem cells, macrophages, alveolar macrophages, ionocytes, pericytes, mesothelial cells, mesenchymal cells, neuroendocrine cells, myofibroblasts, B cells, plasma cells, innate lymphocytes, T cells, monocytes, NK cells, dendritic cells, or PBMCs.

116. The method according to claim 115, wherein the biological sample is serum.

117. The method according to claim 115, wherein the biological sample is plasma.

118. The method according to claim 115, wherein the biological sample is a fibroblast, a basal-like cell, or a PBMC.

119. The method according to any one of claims 111 to 118, wherein the caveolin-1 peptide or its derivative is FTTFTVT (SEQ ID NO: 3).

120. The method according to any one of claims 111 to 118, wherein the caveolin-1 peptide or its derivative is Ac-aaEGKASFTTFTTVTKGSaaNH2 (SEQ ID NO: 4).

121. The method according to any one of claims 1 to 24 or 87 to 96, wherein the subject is a human.

122. The method according to any one of claims 111 to 121, wherein the fibrosis is interstitial lung disease, hepatic fibrosis, renal fibrosis, dermatofibrosis, glomerulonephritis, systemic sclerosis, cardiac fibrosis, myocardial fibrosis, renal fibrosis, cirrhosis, nephrosclerosis, arteriosclerosis, macular degeneration, ocular scarring, cataracts, retina and vitreoretinopathy, Graves' ophthalmopathy, neurofibromatosis, scleroderma, glioblastoma, keloids and hypertrophic scars, peritoneal fibrous disease, chronic obstructive pulmonary disease, postoperative fibroids, diabetic nephropathy, gynecological cancer, myeloproliferative syndrome, myeloid leukemia, myelodysplastic syndrome, inflammatory bowel disease, non-alcoholic fatty liver disease, fibrosarcoma, rheumatoid arthritis, non-alcoholic steatohepatitis, Alport syndrome, or chronic COVID syndrome.

123. The method according to claim 122, wherein the interstitial lung disease is idiopathic pulmonary fibrosis, familial pulmonary fibrosis, idiopathic nonspecific interstitial pneumonia, conventional interstitial lung disease, idiopathic organizing pneumonia, or sarcoidosis.

124. The method according to claim 123, wherein the interstitial lung disease is idiopathic pulmonary fibrosis.

125. The method according to any one of claims 111 to 124, wherein the expression levels of the biomarkers in the first and second biological samples are measured by two-dimensional gel electrophoresis, Western blotting, mass spectrometry, flow cytometry, quantitative RT-PCR, ELISA, and / or lateral flow immunoassay.

126. The method according to any one of claims 111 to 125, wherein an increase in the expression level of MYDGF, PROF1, CALM2, CALR, PPIA, IF5A1, LGALS7, or soluble RAGE shows a favorable reaction to the caveolin-1 peptide or its derivative.

127. The method according to any one of claims 111 to 125, wherein a decrease in the expression level of PDGFRβ, pPDGFRβ, p-mTOR, CALR, IL-11, MMP-2, CXCL7, sCD163, or pSMAD2 / 3 indicates a favorable response to the caveolin-1 peptide or its derivative.

128. The method according to any one of claims 1 to 24 or 111 to 127, wherein the caveolin-1 peptide or a derivative thereof is administered to the subject in a dose of about 0.01 mg / kg to about 250 mg / kg.

129. The method according to claim 128, wherein the caveolin-1 peptide or its derivative is administered to the subject in a dose of about 0.05 mg / kg to about 50 mg / kg.

130. The method according to any one of claims 1 to 24 or 111 to 129, wherein the caveolin-1 peptide or a derivative thereof is administered to the subject by inhalation, intravenous, subcutaneous, oral, intraperitoneal, sublingual, buccal, or intramuscular.

131. The method according to any one of claims 1 to 24 or 111 to 130, wherein the method comprises administering the caveolin-1 peptide or a derivative thereof to the subject once a day, once a week, twice a week, three times a week, five times a week, once every two weeks, or once a month.

132. The method according to any one of claims 1 to 24 or 111 to 131, wherein the second biological sample is obtained from the subject at 1 hour, 3 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 1 week, 2 weeks, 3 weeks, 1 month, 6 months, or 1 year after administration of the caveolin-1 peptide or its derivative.

133. The method according to any one of claims 1 to 24 or 87 to 108, further comprising obtaining one or more additional biological samples after administration of the caveolin-1 peptide or a derivative thereof.

134. The method according to claim 133, wherein the expression level of the biomarker in the one or more additional biological samples is increased compared to the expression level of the biomarker in the first biological sample; and the increase in the expression level of the biomarker in the one or more additional biological samples indicates a favorable reaction to the caveolin-1 peptide or its derivative.

135. The method according to claim 133, wherein the expression level of the biomarker in the one or more additional biological samples is reduced compared to the expression level of the biomarker in the first biological sample; and the reduction in the expression level of the biomarker in the one or more additional biological samples indicates a favorable reaction to the caveolin-1 peptide or its derivative.

136. The method according to any one of claims 111 to 132, wherein the expression level of MYDGF in the second biological sample is increased compared to the first biological sample.

137. The method according to claim 136, wherein the expression level of MYDGF in the second biological sample increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the first biological sample.

138. The method according to any one of claims 111 to 132, wherein the expression level of soluble RAGE in the second biological sample is increased compared to the first biological sample.

139. The method according to claim 138, wherein the expression level of soluble RAGE in the second biological sample increases by about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, or more compared to the first biological sample.

140. The method according to any one of claims 111 to 132, wherein the expression level of pSMAD2 / 3 is reduced compared to the control sample.

141. The method according to claim 140, wherein the expression level of pSMAD2 / 3 is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the control sample.

142. The method according to any one of claims 111 to 132, wherein the expression level of PDGFRβ and / or pPDGFRβ in the second biological sample is reduced in the second biological sample compared to the first biological sample.

143. The method according to claim 142, wherein the expression level of PDGFRβ and / or pPDGFRβ in the second biological sample is reduced by about 10%, about 25%, about 50%, about 75%, about 90%, or more compared to the first biological sample.

144. The method according to any one of claims 111 to 132, wherein the optimal dose of the caveolin-1 peptide or its derivative is determined using the expression level of the biomarker in the second biological sample.

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