ADAMTS14 inhibition

Abstract

Disclosed are methods of treating and preventing fibrosis, and diseases / disorders characterized by fibrosis, through ADAMTS14 inhibition, and agents for use in such methods.

Description

[Technical field]

[0001] This application claims priority to EP21201765.1, filed October 8, 2021, and EP22168132.3, filed April 13, 2022, the contents and elements of which are incorporated herein by reference for all purposes. The present disclosure relates in particular to the field of disease treatment and prevention by inhibition of ADAMTS14. [Background technology]

[0002] Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease with a short life expectancy from the time of diagnosis [1]. IPF causes stiffening of interstitial tissues and reduced gas exchange. Increased stiffness promotes the activation of YAP (Yes-associated protein) and TAZ (transcriptional coactivator with PDZ-binding motifs), two transcriptional coactivators activated by mechanical cues in the environment [4-6]. In response to high stiffness, YAP and TAZ translocate to the nucleus of lung fibroblasts and turn on profibrotic genes. Consistent with this, YAP and TAZ are enriched in the nuclei of fibroblasts in fibrotic areas of IPF lungs [7]. YAP and TAZ are also effectors of the TGFβ signaling pathway and are required for fibroblast activation in the presence of TGFβ [8]. YAP and TAZ activity is suppressed by Hippo pathway kinases through phosphorylation on multiple serine residues [9]. Transfer of fibroblasts overexpressing non-phosphorylatable forms of YAP and TAZ into mouse lungs endowed them with fibrogenic potential, and the mice developed pulmonary fibrosis. [7] In summary, YAP and TAZ are important potential targets for the treatment of IPF.

[0003] ADAMTS14 (a disintegrin and metalloproteinase with thrombospondin motifs 14) is an extracellular enzyme with N-terminal procollagen peptidase activity. It is structurally similar to ADAMTS2 and ADAMTS3 and may have redundant enzymatic activity with ADAMTS2

[10] . Single nucleotide polymorphism (SNP) analysis suggests a role for ADAMTS14 in multiple sclerosis (MS), Achilles tendon, osteoarthritis, and cancer [16-19]. A potential substrate repertoire for ADAMTS14 was proposed by Beckhouche et al. FASEB J. (2016) 30(5):1741-56, including substrates involved in tissue maintenance, remodeling, wound repair, and immune cell function. However, putative ADAMTS14 substrates have pleiotropic roles in various physiological functions, so it is not clear from this study whether and what kind of role ADAMTS14 plays in human disease.

[0004] As noted above, ADAMTS14 has been reported to have aminoprocollagen type I collagenase activity, and for example, Shiomi et al., Pathol Int. (2010) 60(7): 477-496 describes ADAMTS14 as an extracellular matrix degrading proteinase. WO03 / 042379 describes the identification of ADAMTS14 (see, for example, paragraph

[0001] ). US Patent Application Publication No. 2002 / 0119555 identifies a protein having the same sequence as ADAMTS14, designated "53014", as a member of the ADAMTS metalloprotease family, and teaches the use of this protein as a therapeutic agent for the treatment of diseases associated with unwanted extracellular matrix accumulation, such as disorders characterized by fibrosis (see, for example, paragraphs

[0018] ,

[0067] -

[0068] ,

[0350] ). Similarly, U.S. Patent Application Publication No. 2016 / 0331817 describes ADAMTS14 as an antifibrotic agent (see, e.g., paragraph

[0012] ) and teaches administration of ADAMTS14 to a subject for the treatment of a fibrotic disease (see, e.g., claim 2). Summary of the Invention

[0005] In a first aspect, the present disclosure provides an ADAMTS14 inhibitor for use in the treatment or prevention of fibrosis. There is also provided the use of an ADAMTS14 inhibitor in the manufacture of a medicament for treating or preventing fibrosis. Also provided is a method of treating or preventing fibrosis in a subject, comprising administering to the subject a therapeutically or prophylactically effective amount of an ADAMTS14 inhibitor. In some embodiments, the fibrosis is fibrosis of an organ or tissue of the respiratory system, lung, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, an organ or tissue of the cardiovascular system, heart, blood vessels, an organ or tissue of the gastrointestinal system, liver, intestine, small intestine, large intestine, colon, pancreas, skin, eye, an organ or tissue of the nervous system, brain, an organ or tissue of the genitourinary system, kidney, ovary, fallopian tube, an organ or tissue of the musculoskeletal system, muscle tissue, or bone marrow.

[0006] In some embodiments, the fibrosis is selected from the group consisting of pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, asthma, chronic liver disease, liver fibrosis, cirrhosis, non-alcoholic fatty liver disease (NAFLD), steatohepatitis, non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), alcoholic liver disease (ALD), and / or pulmonary fibrosis (PLD). Fatty Liver (AFL), Alcoholic Hepatitis, Alcoholic Steatohepatitis (ASH), Primary Biliary Cirrhosis (PBC), Schistosomal Liver Disease, Hepatocellular Carcinoma (HCC), Hypertrophic Cardiomyopathy (HCM), Dilated Cardiomyopathy (DCM), Atrial Fibrosis, Atrial Fibrillation, Ventricular Fibrosis, Ventricular Fibrillation, Myocardial Fibrosis, Brugada Syndrome, Myocarditis, Endomyocardial Fibrosis, Myocardial Infarction, Fibrous Vascular Disease, Hypertension, Hypertensive Heart Disease, Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC), Atherosclerosis, Chronic Pulmonary Hypertension, AIDS-Related Pulmonary Hypertension, Varicose Veins, Cerebral Infarction, Tubulointerstitial Fibrosis , glomerular fibrosis, renal fibrosis, nephritic syndrome, Alport syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry disease, diabetic nephropathy, chronic glomerulonephritis, nephritis associated with systemic lupus erythematosus, pancreatic fibrosis, chronic pancreatitis, endometriosis, gliosis, Alzheimer's disease, multiple sclerosis, muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), fibrotic myopathy, inflammatory bowel disease (IBD), Crohn's disease, microscopic colitis, primary sclerosing cholangitis (PSC), scleroderma, nephrogenic systemic fibrosis, Dupuytren's contracture, Dermal keloids, Graves' ophthalmopathy, epiretinal fibrosis, retinal fibrosis, subretinal fibrosis, subretinal fibrosis associated with macular degeneration, exudative age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis, postoperative fibrosis of the posterior capsule after cataract surgery, postoperative fibrosis of the bleb after trabeculectomy for glaucoma, conjunctival fibrosis, subconjunctival fibrosis, arthrofibrosis, arthritis, adhesive capsulitis, progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD), fibrotic preneoplastic diseases, fibrotic neoplastic diseases, fibrosis induced by chemical injury,or fibrosis of a disease or condition selected from environmental insult-induced fibrosis, cancer chemotherapy-induced fibrosis, pesticide-induced fibrosis, radiation-induced fibrosis, cancer radiotherapy-induced fibrosis, cancer, hepatocellular carcinoma, gastric cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, vulvar cancer, mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease.

[0007] In some embodiments, the fibrosis is fibrosis of an organ or tissue of the respiratory system, the lungs, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, or bronchi. In some embodiments, the fibrosis is a disease or condition selected from pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in pulmonary disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma. Also provided is an ADAMTS14 inhibitor for use in the treatment or prevention of a disease or condition characterised by fibrosis.

[0008] There is also provided the use of an ADAMTS14 inhibitor in the manufacture of a medicament for treating or preventing a disease or condition characterized by fibrosis. Also provided is a method of treating or preventing a disease or condition characterized by fibrosis in a subject comprising administering to the subject a therapeutically or prophylactically effective amount of an ADAMTS14 inhibitor. In some embodiments, a disease or condition characterized by fibrosis comprises fibrosis of an organ or tissue of the respiratory system, lung, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, an organ or tissue of the cardiovascular system, heart, blood vessels, an organ or tissue of the gastrointestinal system, liver, intestine, small intestine, large intestine, colon, pancreas, skin, eye, an organ or tissue of the nervous system, brain, an organ or tissue of the genitourinary system, ovaries, fallopian tubes, kidneys, an organ or tissue of the musculoskeletal system, muscle tissue, or bone marrow.

[0009] In some embodiments, the disease or condition characterized by fibrosis is pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitial in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, asthma, chronic liver disease, liver fibrosis, cirrhosis, nonalcoholic fatty liver disease (NAFLD), steatohepatitis, nonalcoholic steatohepatitis (NASH), alcoholic fatty liver disease (ALF), pulmonary fibrosis (PL ... Alcoholic Liver Disease (ALD), Alcoholic Fatty Liver (AFL), Alcoholic Hepatitis, Alcoholic Steatohepatitis (ASH), Primary Biliary Cirrhosis (PBC), Schistosomiasis Liver Disease, Hepatocellular Carcinoma (HCC), Hypertrophic Cardiomyopathy (HCM), Dilated Cardiomyopathy (DCM), Atrial Fibrosis, Atrial Fibrillation, Ventricular Fibrosis, Ventricular Fibrillation, Myocardial Fibrosis, Brugada Syndrome, Myocarditis, Endomyocardial Fibrosis, Myocardial Infarction, Fibrous Vascular Disease, Hypertension, Hypertensive Heart Disease, Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC), Atherosclerosis, Chronic Pulmonary Hypertension, AIDS-Related Pulmonary Hypertension, Varicose Veins, Cerebral Infarction, Tubulointerstitial Fibrosis , glomerular fibrosis, renal fibrosis, nephritic syndrome, Alport syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry disease, diabetic nephropathy, chronic glomerulonephritis, nephritis associated with systemic lupus erythematosus, pancreatic fibrosis, chronic pancreatitis, endometriosis, gliosis, Alzheimer's disease, multiple sclerosis, muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), fibrous myopathy, inflammatory bowel disease (IBD), Crohn's disease, microscopic colitis, primary sclerosing cholangitis (PSC), scleroderma, nephrogenic systemic fibrosis, Dupuytren's contracture, dermal ke Lloyd, Graves' ophthalmopathy, epiretinal fibrosis, retinal fibrosis, subretinal fibrosis, subretinal fibrosis associated with macular degeneration, wet age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis, postoperative fibrosis of the posterior capsule after cataract surgery, postoperative fibrosis of the bleb after trabeculectomy for glaucoma, conjunctival fibrosis, subconjunctival fibrosis, arthrofibrosis, arthritis, adhesive capsulitis, progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD), fibrotic preneoplastic diseases, fibrotic neoplastic diseases, fibrosis induced by chemical insult or fibrosis induced by environmental insult,The fibrosis is selected from cancer chemotherapy-induced fibrosis, pesticide-induced fibrosis, radiation-induced fibrosis, cancer radiotherapy-induced fibrosis, cancer, hepatocellular carcinoma, gastric cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, vulvar cancer, mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease.

[0010] In some embodiments, the disease or condition characterized by fibrosis comprises fibrosis of an organ or tissue of the respiratory system, the lung, the bronchioles, the alveoli, the airways, the nasal cavity, the oral cavity, the pharynx, the larynx, the trachea, or the bronchi. In some embodiments, the disease or condition characterized by fibrosis is selected from pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in pulmonary disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma.

[0011] Also provided is a method of inhibiting TGFβ1-mediated signaling in a cell comprising contacting the cell with an ADAMTS14 inhibitor. Also provided is a method of inhibiting YAP-mediated signaling in a cell comprising contacting the cell with an ADAMTS14 inhibitor. Also provided is a method of increasing YAP degradation in a cell comprising contacting the cell with an ADAMTS14 inhibitor. Also provided is a method of inhibiting the production of pro-fibrotic fibroblasts comprising contacting a pro-fibrotic fibroblast precursor cell with an ADAMTS14 inhibitor. Also provided is a method of inhibiting a process mediated by profibrotic fibroblasts comprising contacting a profibrotic fibroblast or a profibrotic fibroblast precursor cell with an ADAMTS14 inhibitor. In some embodiments, the pro-fibrotic fibroblasts are myofibroblasts.

[0012] Also provided is a method of inhibiting the production of myofibroblasts comprising contacting a myofibroblast precursor cell with an ADAMTS14 inhibitor. Also provided is a method of inhibiting a myofibroblast-mediated process comprising contacting a myofibroblast or a myofibroblast precursor cell with an ADAMTS14 inhibitor. In some embodiments, according to various aspects of the present disclosure, the ADAMTS14 inhibitor reduces gene and / or protein expression of ADAMTS14. In some embodiments, according to various aspects of the present disclosure, the ADAMTS14 inhibitor is an inhibitory nucleic acid selected from an siRNA, a dsiRNA, a miRNA, an shRNA, a pri-miRNA, a pre-miRNA, a saRNA, a snoRNA, and an antisense oligonucleotide. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] explanation The present disclosure is based on the inventors' unexpected discovery that ADAMTS14 is a key mediator of the fibrotic response. Antagonism of ADAMTS14 has been shown to inhibit the generation of profibrotic fibroblasts and their activity, resulting in a reduction in correlates of fibrosis. The identification of ADAMTS14 is described in WO03 / 042379. This document includes speculative statements that nucleic acids encoding ADAMTS14, or alternatively its complementary strand, may be useful for the treatment of a wide range of diseases (see, for example, claim 23 of WO03 / 042379, which is dependent on claim 16 and claim 1). However, WO03 / 042379 does not provide any data supporting the use of inhibitors of ADAMTS14 expression / activity for the treatment of any of the specified diseases. WO03 / 042379 further discloses that treatment of fibroblasts with various profibroinflammatory cytokines (including IL-1β, TNFα, and TGFβ) did not modify ADAMTS14 expression (see paragraph

[0076] ). In contrast, ADAMTS1 is disclosed to be an inflammation-related gene whose expression can be induced by IL-1, and the expression of ADAMTS12, ADAMTS4, and ADAMTS5 is similarly disclosed to be upregulated by proinflammatory factors (see paragraph

[0079] ). Thus, WO03 / 042379 suggests that ADAMTS14 is not an effector of the fibroinflammatory process, implying that other fibroinflammatory cytokine-responsive ADAMTS genes (i.e., ADAMTS1, ADAMTS12, ADAMTS4, and ADAMTS5) would be more promising targets for intervention to treat / prevent fibrosis.

[0014] ADAMTS14 The present disclosure particularly relates to inhibition of gene and / or protein expression of ADAM metallopeptidase with thrombospondin type 1 motif 14 (ADAMTS14). The structure and function of ADAMTS14 are described in Coliege et al., J Biol Chem. (2002) 277(8):5756-5766 and Bekhouche and Coliege, Matrix Biol. (2015) 44-46:46-53, both of which are hereby incorporated by reference in their entireties. Alternative splicing of mRNA transcribed from the human ADAMTS14 gene produces four isoforms: isoform A (UniProtKB: Q8WXS8-1, v2; SEQ ID NO: 1); isoform B (UniProtKB: Q8WXS8-2; SEQ ID NO: 2), which lacks the amino acid sequence corresponding to positions 1 to 67 of SEQ ID NO: 1; isoform C (UniProtKB: Q8WXS8-3; SEQ ID NO: 3), which lacks the amino acid sequence corresponding to positions 1 to 67 of SEQ ID NO: 1 and has an insertion of "MQG" after the "G" at the position corresponding to position 368 of SEQ ID NO: 1; and isoform D (UniProtKB: Q8WXS8-4; SEQ ID NO: 4), which has an insertion of "MQG" after the "G" at the position corresponding to position 368 of SEQ ID NO: 1.

[0015] The 1223 amino acid sequence of human ADAMTS14 isoform A includes an N-terminal signal peptide (shown in SEQ ID NO:5, positions 1 to 22 of SEQ ID NO:1), followed by a 230 amino acid propeptide (shown in SEQ ID NO:6, positions 23 to 252 of SEQ ID NO:1), and a mature protein region (shown in SEQ ID NO:7, positions 253 to 1223 of SEQ ID NO:1). The mature protein region includes an N-terminal peptidase M12B domain (positions 259-460 of SEQ ID NO:1, as shown in SEQ ID NO:10), a disintegrin domain (positions 461-551 of SEQ ID NO:1, as shown in SEQ ID NO:11), four TSP1-type repeats (positions 552-607, 847-907, 908-967, and 968-1022 of SEQ ID NO:1, as shown in SEQ ID NO:13, 14, 15, and 16, respectively) with a spacer domain provided between TSP1-type repeats 1 and 2 (positions 730-846 of SEQ ID NO:1, as shown in SEQ ID NO:17), a PLAC domain (positions 1059-1097 of SEQ ID NO:1, as shown in SEQ ID NO:18), and a C-terminal proline-rich region (positions 1100-1223 of SEQ ID NO:1, as shown in SEQ ID NO:19). ADAMTS14 has been reported to exhibit aminoprocollagen type I collagenase activity (Colige et al., supra).

[0016] In the experimental examples of the present disclosure, the inventors demonstrate that ADAMTS14 is involved in the nuclear localization of YAP and the regulation of the levels and phosphorylation of YAP and TAZ, as well as YAP target genes and profibrotic mediators CTGF and CYR61 in fibroblasts. ADAMTS14 is involved in TGFβ1-induced upregulation of the expression of profibrotic and metaplastic differentiation genes. ADAMTS14 has also been shown to be involved in the TGFβ1-induced phosphorylation of SMAD2 and the TGFβ1-induced localization of SMAD2 to the nucleus. ADAMTS14 has also been shown to be important for the formation of α-SMA fibers in fibroblasts and collagen deposition by fibroblasts in response to TGFβ1. As used herein, reference to "ADAMTS14" encompasses human ADAMTS14 isoforms A, B, C, and D, homologues of human ADAMTS14 isoforms A, B, C, and D (i.e., encoded by the genome of a non-human animal), and variants thereof. Homologs of human ADAMTS14 isoforms A, B, C, or D can be from any animal. In some embodiments, homologs of human ADAMTS14 isoforms A, B, C, or D can be from a mammal. In some embodiments, the mammal can be a non-human mammal, such as a primate (e.g., a non-human primate, such as an animal of the genus Macaca (e.g., Macaca fascicularis, Macaca mulatta), such as a non-human hominidae (e.g., Pan troglodytes). In some embodiments, the mammal can be a rabbit, a guinea pig, a rat, a mouse, or an animal of the order Rodentia, a cat, a dog, a pig, a sheep, a goat, an animal of the order Bos (e.g., a cow), an animal of the family Equidae (e.g., a horse), or a donkey.

[0017] Homologs of human ADAMTS14 isoforms (e.g., isoforms A, B, C, or D) may be characterized as having 70% or more amino acid sequence identity with the amino acid sequence of the related isoform, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity. Variants of human ADAMTS14 isoforms (e.g., isoforms A, B, C, or D) may be characterized as having 70% or more amino acid sequence identity with the related isoform, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more amino acid sequence identity. In some embodiments, ADAMTS14 according to the present disclosure comprises or consists of an amino acid sequence having 70% or more amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO:1, 2, 3, or 4.

[0018] In some embodiments, ADAMTS14 according to the present disclosure comprises an amino acid sequence having 70% or more amino acid sequence identity, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity, to the amino acid sequence of SEQ ID NO: 8 or 9. ADAMTS14 inhibitors Aspects of the present disclosure relate to inhibitors of ADAMTS14. "ADAMTS14 inhibitor" refers to any agent capable of inhibiting ADAMTS14 expression and / or function. That is, an inhibitor of ADAMTS14 includes an agent that reduces the level of ADAMTS14 gene and / or protein expression, as well as an agent that reduces the activity of ADAMTS14. It will be understood that reduction in the preceding sentence refers to the level of expression / activity observed in the absence of such inhibition. In the present disclosure, an inhibitor of ADAMTS14 may also be referred to as an "antagonist" of ADAMTS14, and similarly, inhibition of ADAMTS14 may be referred to as "antagonism" of ADAMTS14.

[0019] As used herein, an "activity" or "function" of or mediated by ADAMTS14 may refer to cleavage of aminoprocollagen type I, nuclear localization of YAP, nuclear localization of TAZ, nuclear localization of SMAD2, phosphorylation of SMAD2, upregulation of expression of one or more genes involved in fibrosis and / or metaplastic differentiation, formation of αSMA fibers and / or collagen I deposition (e.g., in / by fibroblasts). In some embodiments, ADAMTS14 inhibitors according to the present disclosure exhibit one or more of the following properties: reducing expression (e.g., gene and / or protein expression) of ADAMTS14; Reduces levels of the RNA encoding ADAMTS14 Reducing the transcription of nucleic acid encoding ADAMTS14, Increases the degradation of the RNA that codes for ADAMTS14, Reduces levels of ADAMTS14 protein, reducing post-transcriptional processing (e.g., splicing, translation, post-translational processing) of RNA encoding ADAMTS14; Increases the degradation of ADAMTS14 protein, Reduces levels of correlates of ADAMTS14 activity,

[0020] Reduces the cleavage of type I aminoprocollagen reducing TGFβ1-mediated upregulation of expression of one or more genes involved in fibrosis and / or metaplastic differentiation (e.g., one or more genes selected from ADAMTS14, ACTA2, COL1A1, FN1, FOXJ1, KRT5, MUC5AC, MUC5B, SCGB1A1, CTGF, CYR61, MMP7, and SOX2; e.g., in fibroblasts or epithelial cells); Decreasing the levels of YAP (e.g., in fibroblasts); Reduces nuclear localization of YAP (e.g., in fibroblasts); Increase the degradation of YAP (e.g., in fibroblasts), increasing the level of phosphorylated YAP (e.g., YAP phosphorylated at S397; e.g., in fibroblasts); reducing the levels of proteins encoded by genes whose expression is upregulated by YAP (e.g., in fibroblasts); Decreasing the levels of TAZ (e.g., in fibroblasts), Reduces nuclear localization of TAZ (e.g., in fibroblasts); increasing the levels of phosphorylated TAZ (e.g., TAZ phosphorylated at S89; e.g., in fibroblasts); reducing the levels of proteins encoded by genes whose expression is upregulated by TAZ (e.g., in fibroblasts);

[0021] reducing levels of CTGF (e.g., in fibroblasts); Reducing levels of CYR61 (e.g., in fibroblasts); Reduce TGFβ1-mediated signaling (e.g., in fibroblasts); reducing TGFβ1-mediated upregulation of levels of phosphorylated SMAD2 (e.g., in fibroblasts); reducing TGFβ1-mediated upregulation of SMAD2 localization to the nucleus (e.g., in fibroblasts); Reduce TGFβ1-mediated upregulation of levels of αSMA (e.g., in fibroblasts); and / or reducing TGFβ1-mediated production of components of the extracellular matrix (e.g., type 1 collagen I; e.g., in / by fibroblasts); Reduces extracellular matrix stiffening (e.g., of the extracellular matrix produced by fibroblasts or epithelial cells in culture in vitro); and Reducing stiffening of tissues / organs (e.g., lung, liver, skin, kidney), including fibrosis.

[0022] It will be understood that a given ADAMTS14 inhibitor may exhibit more than one of the properties listed in the preceding paragraph.A given ADAMTS14 inhibitor can be evaluated for the properties listed in the preceding paragraph using a suitable assay.For example, the assay can be, for example, an in vitro assay, optionally a cell-based assay or a cell-free assay.In some embodiments, the assay can be, for example, an in vivo assay, i.e., carried out in a non-human animal.In some embodiments, the assay can be, for example, an ex vivo assay, i.e., carried out using cells / tissues / organs obtained from a subject.

[0023] When the assays are cell-based assays, they may include treating cells with a given agent to determine whether the agent exhibits one or more of the listed properties. The assays may employ species labeled with detectable entities to facilitate their detection. The assays may include treating cells separately with a range of amounts / concentrations (e.g., a dilution series) of a given agent, followed by evaluating the listed properties. It will be appreciated that the cells employed in such are preferably cells that express ADAMTS14, such as fibroblasts (e.g., lung fibroblasts). According to various aspects and embodiments of the present disclosure, one or more fibroblasts can reside or originate from tissue or organ of interest.In some embodiments, one or more fibroblasts can reside or originate from lung, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, bronchus, heart, kidney, liver, skeletal muscle, blood vessel, eye, skin, pancreas, intestine, small intestine, large intestine, colon, joint, brain, or bone marrow.In some embodiments, one or more fibroblasts can reside or originate from lung.

[0024] In some embodiments, the assay may include treating cells to upregulate ADAMTS14 expression and / or activity. For example, the assay may include treating cells with TGFβ1 (e.g., at a final concentration of 5 ng / ml) or with an IPF-related cytokine cocktail (IPF-RC, e.g., as described in Schruf et al. FASEB J. (2020) 34(6):7825-7846; e.g., at a 1:100 dilution). Analysis of the results of such assays may include determining the concentration at which 50% of the maximum level of the relevant activity is reached. The concentration of a given agent at which 50% of the maximum level of the relevant activity is reached may be referred to as the "half maximal effective concentration" of the agent in relation to the relevant activity, which is referred to as the "EC 50 By way of example, the EC of a given agent for increasing the degradation of RNA encoding ADAMTS14 may be referred to as 50 may be the concentration of an agent at which 50% of maximal degradation of RNA encoding ADAMTS14 is achieved. Depending on the characteristics, EC 50 is the "half maximal inhibitory concentration" or "IC 50 ", which is the concentration of an agent at which 50% of the maximal level of inhibition of a given property is observed. By way of example, the IC of a given agent for reducing gene expression of ADAMTS14 is 50 may be the concentration of an agent at which 50% of the maximal level of inhibition of ADAMTS14 expression is achieved.

[0025] ADAMTS14 inhibitors that can reduce gene expression of ADAMTS14 and / or reduce the level of RNA encoding ADAMTS14 and / or reduce the transcription of nucleic acid encoding ADAMTS14 and / or increase the degradation of RNA encoding ADAMTS14 can be identified using an assay that includes detecting and / or quantifying the level of RNA encoding ADAMTS14. Such an assay can include quantifying the RNA encoding ADAMTS14 by techniques well known to those skilled in the art such as RT-qPCR, Northern blot, etc. The method can employ primers and / or probes for detecting and / or quantifying the RNA encoding ADAMTS14. Such an assay can include contacting cells expressing ADAMTS14 (e.g., fibroblasts) with a putative ADAMTS14 inhibitor in in vitro culture, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the change in the level of the RNA encoding ADAMTS14 of interest to be observed) measuring the level of the RNA encoding ADAMTS14. Such assays may further include a step of comparing the level of RNA encoding ADAMTS14 in cells treated with a putative ADAMTS14 inhibitor with the level of RNA encoding ADAMTS14 detected in control conditions in which cells of the same type are subjected to the same conditions, except that instead of being treated with the putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect the level of RNA encoding ADAMTS14.

[0026] Decreased transcription of nucleic acid encoding ADAMTS14 can be the result of inhibition of the association and / or activity of factors required for transcription of DNA encoding ADAMTS14. Increased degradation of RNA encoding ADAMTS14 can be the result of increased enzymatic degradation of RNA encoding ADAMTS14, for example as a result of RNA interference (RNAi), and / or decreased stability of RNA encoding ADAMTS14. As used herein, "contacting" a cell with a given agent (e.g., a putative ADAMTS14 inhibitor) may include applying the agent to the cell and / or mixing the agent with the cell. In some embodiments, the putative ADAMTS14 inhibitor is provided to the cell in combination with one or more additional agents to promote the introduction of the putative ADAMTS14 inhibitor into the cell and / or promote uptake of the putative ADAMTS14 inhibitor by the cell. For example, in embodiments in which the putative ADAMTS14 inhibitor is or is encoded by one or more nucleic acids, the cell may be contacted with the nucleic acid and an agent to promote the introduction of the nucleic acid into the cell, for example, by transfection or transduction. In some embodiments, a putative ADAMTS14 inhibitor may be assessed for its ability to reduce gene expression of ADAMTS14 (eg, in fibroblasts), as described in Example 1 herein.

[0027] ADAMTS14 inhibitors that can reduce post-transcriptional processing of RNA encoding ADAMTS14 (e.g., reduce normal splicing of pre-mRNA encoding ADAMTS14) can be identified using an assay that includes detecting and / or quantifying the level of RNA (e.g., mature mRNA) encoding one or more isoforms of ADAMTS14. Such an assay can include quantifying RNA (e.g., mature mRNA) encoding one or more isoforms of ADAMTS14 by RT-qPCR. The method can employ primers and / or probes for detecting and / or quantifying mature mRNA produced by standard splicing of pre-mRNA transcribed from the gene encoding ADAMTS14 and / or primers and / or probes for detecting and / or quantifying mature mRNA produced by alternative splicing of pre-mRNA transcribed from the gene encoding ADAMTS14. The mature mRNA produced by standard splicing of the pre-mRNA transcribed from the gene encoding ADAMTS14 may be a mature mRNA encoding the major isoform produced by the expression of the gene encoding ADAMTS14. The major isoform may be the most commonly produced / detected isoform. For example, the mature mRNA produced by standard splicing of the pre-mRNA transcribed from human ADAMTS14 may be a mature mRNA encoding human ADAMTS14 isoform A (i.e., having the amino acid sequence shown in SEQ ID NO: 1). The mature mRNA produced by alternative splicing of the pre-mRNA transcribed from the gene encoding ADAMTS14 may be a mature mRNA encoding an isoform other than the major isoform produced by the expression of the gene encoding ADAMTS14.For example, the mature mRNA produced by alternative splicing of pre-mRNA transcribed from human ADAMTS14 may be a mature mRNA encoding an isoform of human ADAMTS14 other than isoform A (i.e., having an amino acid sequence that is not identical to SEQ ID NO: 1); for example, a mature mRNA encoding human ADAMTS14 isoform B, C, or D. Such an assay may include contacting cells expressing ADAMTS14 (e.g., fibroblasts) in in vitro culture with a putative ADAMTS14 inhibitor, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for an effect on post-transcriptional processing of RNA encoding ADAMTS14 of interest to be observed (e.g., splicing of pre-mRNA encoding ADAMTS14)), measuring the level of mature mRNA encoding one or more isoforms of ADAMTS14. Such assays may further include a step of comparing the levels of mature mRNA encoding one or more isoforms of ADAMTS14 in cells treated with a putative ADAMTS14 inhibitor with the levels of mature mRNA encoding one or more isoforms of ADAMTS14 detected in control conditions in which cells of the same type are subjected to the same conditions, except that instead of being treated with a putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect splicing of the pre-mRNA encoding ADAMTS14.

[0028] A reduction in the level of standard splicing of pre-mRNA encoding ADAMTS14 may be the result of inhibition of the association and / or activity of factors required for standard splicing. A reduction in translation of mRNA encoding ADAMTS14 may be the result of inhibition of the association and / or activity of factors required for translation. A reduction in post-translational processing (e.g., enzymatic processing, folding) of ADAMTS14 may be the result of inhibition of the association and / or activity of factors required for post-translational processing of ADAMTS14. An increase in degradation of ADAMTS14 protein may be the result of increased enzymatic (e.g., protease-mediated) degradation of ADAMTS14 protein.

[0029] ADAMTS14 inhibitors that can reduce the level of ADAMTS14 protein and / or increase the degradation of ADAMTS14 protein and / or reduce the translation of mRNA encoding ADAMTS14 can be identified using assays that include detecting the level of ADAMTS14 protein, using techniques well known to those skilled in the art, such as antibody / reporter-based methods (Western blot, ELISA, immunohisto / cytochemistry, etc.). The method can employ an antibody specific to ADAMTS14. Such an assay can include contacting cells expressing ADAMTS14 (e.g., fibroblasts) with a putative ADAMTS14 inhibitor in in vitro culture, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the change in the level of the ADAMTS14 protein of interest to be observed) measuring the level of ADAMTS14 protein. Such assays may further include a step of comparing the level of ADAMTS14 protein in cells treated with a putative ADAMTS14 inhibitor with the level of ADAMTS14 protein detected in control conditions in which the same type of cells are subjected to the same conditions, except that instead of being treated with the putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect the level of ADAMTS14 protein.

[0030] A decrease in the level of ADAMTS14 protein can be the result of, for example, a decrease in the level of RNA encoding ADAMTS14, a decrease in post-transcriptional processing of RNA encoding ADAMTS14, or an increase in degradation of the ADAMTS14 protein. In some embodiments, a putative ADAMTS14 inhibitor can be assessed for its ability to reduce levels of ADAMTS14 protein (eg, in fibroblasts), as described in Example 1 herein. ADAMTS14 inhibitors that can reduce the level of ADAMTS14 function (e.g., the ADAMTS14 function described above) can be identified using an assay that includes detecting the level of a related function. Detecting the level of a given function can include detecting and / or quantifying a correlate of the function. By way of example, detecting the level of cleavage of type I aminoprocollagen can include detecting and / or quantifying the level of cleaved and / or uncleaved type I aminoprocollagen. Such an assay can include contacting cells expressing ADAMTS14 (e.g., fibroblasts) with a putative ADAMTS14 inhibitor in in vitro culture, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the level of the related function of interest and / or its correlate to be reduced) measuring the level of the related function and / or its correlate. Such assays may further include a step of comparing the level of ADAMTS14 function in cells treated with a putative ADAMTS14 inhibitor with the level of ADAMTS14 function detected in control conditions in which cells of the same type are subjected to the same conditions, except that instead of being treated with the putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect the level of the relevant function of ADAMTS14.

[0031] ADAMTS14 inhibitors that can reduce the cleavage of type I aminoprocollagen can be identified using an assay that includes detecting and / or quantifying the level of cleaved and / or uncleaved type I aminoprocollagen. Such an assay can include contacting cells expressing ADAMTS14 (e.g., fibroblasts) with a putative ADAMTS14 inhibitor in in vitro culture, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the observed reduction in the cleavage of type I aminoprocollagen and / or its correlates) measuring the level of cleaved and / or uncleaved type I aminoprocollagen. Such an assay can further include comparing the level of cleavage of type I aminoprocollagen in cells treated with the putative ADAMTS14 inhibitor with the level detected in a control condition in which the same type of cells are subjected to the same conditions, except that instead of being treated with the putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect the level of cleavage of type I aminoprocollagen.

[0032] ADAMTS14 inhibitors that can reduce the upregulation of expression of one or more genes involved in fibrosis and / or metaplastic differentiation (e.g., one or more genes selected from ADAMTS14, ACTA2, COL1A1, FN1, FOXJ1, KRT5, MUC5AC, MUC5B, SCGB1A1, CTGF, CYR61, MMP7, and SOX2; e.g., in fibroblasts or epithelial cells) in response to stimulation with IPF-RC can be identified using an assay that includes detecting and / or quantifying the level of expression of the relevant gene. Such an assay can include contacting a cell (e.g., fibroblast) that expresses ADAMTS14 with a putative ADAMTS14 inhibitor and TGFβ1 under in vitro culture, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the observed reduction in the level of expression of the relevant gene of interest), for example, by detecting and / or quantifying the level of RNA encoding the product of the relevant gene. Such assays may further include a step of comparing the level of expression of the associated gene in cells treated with a putative ADAMTS14 inhibitor with the level detected in a control condition in which cells of the same type are subjected to the same conditions, except that instead of being treated with a putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect the level of expression of the associated gene.

[0033] Genes whose expression is upregulated by YAP / TAZ are described, for example, in Moya and Halder, Nat. Rev. Mol. Cell Biol. (2019) 20:211-226 (hereby incorporated by reference in its entirety), and include, for example, CTGF, CYR61, BIRC5, AREG, AMOTL1, and ANKRD1. In some embodiments, as described in Example 1 herein, a putative ADAMTS14 inhibitor may be evaluated for its ability to reduce upregulation of expression of one or more genes involved in fibrosis and / or metaplastic differentiation (e.g., one or more genes selected from ADAMTS14, ACTA2, COL1A1, FN1, FOXJ1, KRT4, MUC5AC, MC5B, SCGB1A1, CTGF, CYR61, MMP7, and SOX2; e.g., in fibroblasts or epithelial cells) in response to stimulation with IPF-RC.

[0034] ADAMTS14 inhibitors that can reduce the level of a given protein (e.g., YAP, TAZ, SMAD2, collagen I, αSMA, proteins encoded by genes whose expression is upregulated by YAP / TAZ), or a specific post-translationally modified form thereof (e.g., phosphorylated SMAD2), can be identified using an assay that includes detecting and / or quantifying the level of the relevant protein / its post-translationally modified form, using techniques well known to those skilled in the art, such as, for example, antibody / reporter-based methods (Western blot, ELISA, immunohisto / cytochemistry, etc.). The method may employ an antibody specific for the relevant protein or its post-translationally modified form. Such an assay may include contacting cells expressing ADAMTS14 (e.g., fibroblasts) in in vitro culture with a putative ADAMTS14 inhibitor and optionally TGFβ1, and thereafter (e.g., after a suitable period of time, i.e., a period of time sufficient for the change in the level of the relevant protein / its post-translationally modified form of interest to be observed), measuring the level of the relevant protein / its post-translationally modified form. Such assays may further include a step of comparing the levels of the associated protein / its post-translationally modified forms in cells treated with a putative ADAMTS14 inhibitor with the levels detected in control conditions in which the same type of cells are subjected to the same conditions, except that instead of being treated with a putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect the levels of the associated protein / its post-translationally modified forms.

[0035] In some embodiments, as described in Example 1 herein, a putative ADAMTS14 inhibitor may be assessed for its ability to reduce levels of YAP, TAZ, SMAD2, collagen I, αSMA, phosphorylated SMAD2, a protein encoded by a gene whose expression is upregulated by YAP, or a protein encoded by a gene whose expression is upregulated by TAZ (e.g., in fibroblasts, e.g., following stimulation with TGFβ1). ADAMTS14 inhibitors that can reduce TGFβ1-mediated signaling can be identified using assays that include detecting and / or quantifying the level of a correlate of TGFβ1-mediated signaling, for example, using techniques well known to those skilled in the art, such as antibody / reporter-based methods (Western blot, ELISA, immunohisto / cytochemistry, etc.). The method can employ an antibody specific for a correlate of TGFβ1-mediated signaling, such as phosphorylated SMAD2. Such an assay can include contacting cells expressing ADAMTS14 (e.g., fibroblasts) with a putative ADAMTS14 inhibitor and TGFβ1 in in vitro culture, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the inhibition of TGFβ1-mediated signaling to be observed), measuring the level of the correlate of TGFβ1-mediated signaling. Such assays may further include a step of comparing the levels of correlates of TGFβ1-mediated signaling in cells treated with a putative ADAMTS14 inhibitor with the levels detected in control conditions in which the same type of cells are subjected to the same conditions, except that instead of being treated with a putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect TGFβ1-mediated signaling.

[0036] In some embodiments, a putative ADAMTS14 inhibitor may be evaluated for its ability to reduce TGFβ1-mediated signaling (eg, in fibroblasts), as described in Example 1 herein. ADAMTS14 inhibitors that can increase or decrease the proportion of a given protein in its specific post-translationally modified form can be identified using an assay that includes detecting and / or quantifying the levels of the associated protein and / or its post-translationally modified form, for example using techniques well known to those skilled in the art, such as antibody / reporter-based methods (Western blot, ELISA, immunohisto / cytochemistry, etc.). The method can employ an antibody specific for the associated protein and an antibody specific for its post-translationally modified form. Such an assay can include contacting cells expressing ADAMTS14 (e.g., fibroblasts) in in vitro culture with a putative ADAMTS14 inhibitor and optionally TGFβ1, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the change in the proportion of the given protein in its specific post-translationally modified form to be observed), measuring the level of the associated protein and the level of its post-translationally modified form. Such an assay can further include determining the proportion of the associated protein in its post-translationally modified form. Such assays may further include a step of comparing the proportion of associated proteins in their post-translationally modified form determined in cells treated with a putative ADAMTS14 inhibitor with the proportion determined in control conditions in which cells of the same type are subjected to the same conditions, except that instead of being treated with a putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect the proportion of associated proteins in their post-translationally modified form.

[0037] ADAMTS14 inhibitors that can reduce the nuclear localization of a given protein or its specific post-translational modified form (e.g., YAP, TAZ, SMAD2, phosphorylated SMAD2) and / or reduce the percentage of a given protein or its specific post-translational modified form that is localized in the nucleus can be identified using assays that include detecting, quantifying, and / or determining the percentage of the relevant protein / its post-translational modified form that is localized in the nucleus, using techniques well known to those skilled in the art, such as, for example, antibody / reporter-based methods (Western blot, ELISA, immunohisto / cytochemistry, etc.). The subcellular localization of a given protein / its post-translational modified form can be analyzed, for example, by immunocytochemistry or Western blot of extracts prepared from various cell fractions, and can employ organelle (e.g., nuclear) markers and / or labeling proteins of known subcellular localization. Such an assay may include contacting cells expressing ADAMTS14 (e.g., fibroblasts) in in vitro culture with a putative ADAMTS14 inhibitor and, optionally, with TGFβ1, and then (e.g., after a suitable period of time, i.e., a period of time sufficient for the observed change in the subcellular localization of the given protein / its post-translational modified form) detecting and / or quantifying the given protein / its post-translational modified form that is localized in the nucleus and / or that is not localized in the nucleus (e.g., that is localized in the cytoplasm). Such an assay may further include determining the proportion of the relevant protein / its post-translational modified form that is localized in the nucleus. Such assays may further include a step of comparing the percentage of associated proteins / their post-translationally modified forms that are localized to the nucleus in cells treated with a putative ADAMTS14 inhibitor with the percentage determined in control conditions in which cells of the same type are subjected to the same conditions, except that instead of being treated with a putative ADAMTS14 inhibitor, they are not treated or are otherwise treated with a negative control agent known not to affect localization of associated proteins / their post-translationally modified forms to the nucleus.

[0038] In some embodiments, the ADAMTS14 inhibitor according to the present disclosure can reduce the stiffening of extracellular matrix. In some embodiments, the extracellular matrix can be produced by fibroblasts or epithelial cells in vitro in culture. In some embodiments, the ADAMTS14 inhibitor can reduce the stiffening of tissue or organ. In some embodiments, the tissue / organ can be or be derived from the lung, liver, skin, or kidney. In some embodiments, the tissue / organ is a tissue / organ that contains fibrosis (i.e., in some embodiments, the tissue / organ is fibrotic).

[0039] Stiffening of the extracellular matrix, or stiffening of a given tissue / organ, can be assessed, for example, by determining Young's modulus, as described, for example, in Akhmanova et al., Stem Cells Int. (2015) 2015:167025, which is hereby incorporated by reference in its entirety. Methods for assessing Young's modulus are summarized, for example, in Table 2 of Akhmanova et al., and included analysis by ultrasound elastography, atomic force microscopy, or millimeter indentation.

[0040] In some embodiments, an ADAMTS14 inhibitor according to the present disclosure exhibits an activity in a given assay that is less than 1-fold, e.g., ≦0.99-fold, ≦0.95-fold, ≦0.9-fold, ≦0.85-fold, ≦0.8-fold, ≦0.75-fold, ≦0.7-fold, ≦0.65-fold, ≦0.6-fold, ≦0.55-fold, ≦0.5-fold, ≦0.45-fold, ≦0.4-fold, ≦0.35-fold, ≦0.3-fold, ≦0.25-fold, ≦0.2-fold, ≦0.15-fold, ≦0.1-fold, ≦0. 0.05-fold, or ≦0.01-fold, the expression of the gene encoding ADAMTS14 may be reduced; the level of RNA encoding ADAMTS14 may be reduced; the transcription of nucleic acid encoding ADAMTS14 may be reduced; the level of ADAMTS14 protein may be reduced; the post-transcriptional processing of RNA encoding ADAMTS14 may be reduced; the normal splicing of pre-mRNA encoding ADAMTS14 may be reduced; the translation of mRNA encoding ADAMTS14 may be reduced; may reduce the level of a correlate of sexual function / may reduce the cleavage of type I aminoprocollagen / may reduce TGFβ1-mediated upregulation of expression of one or more genes involved in fibrosis and / or metaplastic differentiation / may reduce the level of YAP / may reduce the nuclear localization of YAP / may reduce the level of a protein encoded by a gene whose expression is upregulated by YAP / may reduce the level of TAZ / may reduce the nuclear localization of TAZ / may reduce the level of a protein encoded by a gene whose expression is upregulated by TAZ may reduce levels of CTGF; may reduce levels of CYR61; may reduce TGFβ1-mediated signaling; may reduce upregulation of levels of phosphorylated SMAD2 in response to stimulation with TGFβ1; may reduce TGFβ1-mediated upregulation of SMAD2 localization to the nucleus; may reduce TGFβ1-mediated upregulation of levels of αSMA; may reduce TGFβ1-mediated production of components of the extracellular matrix (e.g., type 1 collagen); may reduce extracellular matrix stiffening; may reduce stiffening of tissues or organs.

[0041] In some embodiments, an ADAMTS14 inhibitor according to the present disclosure inhibits ADAMTS14 activity in a given assay by less than 100%, e.g., to one of: ≦99%, ≦95%, ≦90%, ≦85%, ≦80%, ≦75%, ≦70%, ≦65%, ≦60%, ≦55%, ≦50%, ≦45%, ≦40%, ≦35%, ≦30%, ≦25%, ≦20%, ≦15%, ≦10%, ≦5%, or ≦1%, of the level observed in the absence of the ADAMTS14 inhibitor or in the presence of an equal amount of a control agent known to have no such inhibitory activity. may reduce the expression of a gene encoding MTS14 / may reduce the level of an RNA encoding ADAMTS14 / may reduce the transcription of a nucleic acid encoding ADAMTS14 / may reduce the level of an ADAMTS14 protein / may reduce post-transcriptional processing of an RNA encoding ADAMTS14 / may reduce normal splicing of a pre-mRNA encoding ADAMTS14 / may reduce the translation of an mRNA encoding ADAMTS14 / may reduce the level of a correlate of ADAMTS14 activity / may reduce the cleavage of type I aminoprocollagen / may reduce TGFβ1-mediated upregulation of the expression of one or more genes involved in fibrosis and / or metaplastic differentiation / may reduce the level of YAP / may reduce the nuclear localization of YAP / may reduce the level of a protein encoded by a gene whose expression is upregulated by YAP / may reduce the level of TAZ / may reduce the nuclear localization of TAZ / may reduce the level of a protein encoded by a gene whose expression is upregulated by TAZ / CTGF may reduce the levels of CYR61; may reduce TGFβ1-mediated signaling; may reduce upregulation of the levels of phosphorylated SMAD2 in response to stimulation with TGFβ1; may reduce TGFβ1-mediated upregulation of SMAD2 localization to the nucleus; may reduce TGFβ1-mediated upregulation of the levels of αSMA; may reduce TGFβ1-mediated production of components of the extracellular matrix (e.g., type 1 collagen); may reduce extracellular matrix stiffening; may reduce stiffening of tissues or organs.

[0042] Preferred levels of reduction according to the preceding two paragraphs are a reduction of less than 0.5 times / ≦50%, for example to one of less than 0.4 times / ≦40%, less than 0.3 times / ≦30%, less than 0.2 times / ≦20%, less than 0.15 times / ≦15%, or less than 0.1 times / ≦10%. In some embodiments, an ADAMTS14 inhibitor according to the present disclosure exhibits a cytotoxic effect in a given assay that is greater than 1-fold, e.g., ≧1.01-fold, ≧1.02-fold, ≧1.03-fold, ≧1.04-fold, ≧1.05-fold, ≧1.1-fold, ≧1.2-fold, ≧1.3-fold, ≧1.4-fold, ≧1.5-fold, ≧1.6-fold, ≧1.7-fold, ≧1.8-fold, ≧1.9-fold, It may increase the degradation of RNA encoding ADAMTS14 / may increase the degradation of ADAMTS14 protein / may increase the degradation of YAP / may increase the levels of phosphorylated YAP (e.g., YAP phosphorylated at S397) / may increase the degradation of TAZ / may increase the levels of phosphorylated TAZ (e.g., TAZ phosphorylated at S89) by one of the following: ≧2-fold, ≧3-fold, ≧4-fold, ≧5-fold, ≧6-fold, ≧7-fold, ≧8-fold, ≧9-fold, or ≧10-fold.

[0043] In some embodiments, the ADAMTS14 inhibitor according to the present disclosure inhibits or silences the expression of the gene encoding ADAMTS14.In some embodiments, the ADAMTS14 inhibitor according to the present disclosure inhibits or silences the expression of ADAMTS14 at the protein level.As used herein, the expression of a given gene / protein can be considered to be "inhibited" or "silenced" if the level of expression is reduced to less than 0.1-fold / ≦10% of the level observed in the absence of the putative ADAMTS14 inhibitor or in the presence of the same amount of a control agent that is known not to inhibit the expression of related genes / proteins. In a preferred embodiment, an ADAMTS14 inhibitor according to the present disclosure inhibits ADAMTS14 gene and / or protein expression in a given assay by more than 50%, e.g., ≧60%, ≧61%, ≧62%, ≧63%, ≧64%, ≧65%, or more than 50% of ADAMTS14 gene and / or protein expression observed in the absence of the ADAMTS14 inhibitor or in the presence of an equal amount of a control agent known not to inhibit ADAMTS14 gene and / or protein expression. ≧66%, ≧67%, ≧68%, ≧69%, ≧70%, ≧71%, ≧72%, ≧73%, ≧74%, ≧75%, ≧76%, ≧77%, ≧78%, ≧79%, ≧80%, ≧81%, ≧82%, ≧83%, ≧84%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100% inhibition.

[0044] In a preferred embodiment, an ADAMTS14 inhibitor according to the present disclosure inhibits ADAMTS14 gene expression in a given assay (e.g., the assay described in Example 1 herein) that is greater than 50%, e.g., ≧60%, ≧61%, ≧62%, ≧70%, ≧71%, ≧72%, ≧73%, ≧74%, ≧75%, ≧76%, ≧77%, ≧78%, ≧79 ... ≧63%, ≧64%, ≧65%, ≧66%, ≧67%, ≧68%, ≧69%, ≧70%, ≧71%, ≧72%, ≧73%, ≧74%, ≧75%, ≧76%, ≧77%, ≧78%, ≧79%, ≧80%, ≧81%, ≧82%, ≧83%, ≧84%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100% inhibition. In a preferred embodiment, an ADAMTS14 inhibitor according to the present disclosure inhibits ADAMTS14 protein expression in a given assay (e.g., the assay described in Example 1 herein) that is greater than 50%, e.g., ≧60%, ≧61%, ≧62%, or greater than 50% of ADAMTS14 protein expression (e.g., as determined by ELISA) observed in the absence of the ADAMTS14 inhibitor or in the presence of an equal amount of a control agent known not to inhibit ADAMTS14 protein expression. ≧63%, ≧64%, ≧65%, ≧66%, ≧67%, ≧68%, ≧69%, ≧70%, ≧71%, ≧72%, ≧73%, ≧74%, ≧75%, ≧76%, ≧77%, ≧78%, ≧79%, ≧80%, ≧81%, ≧82%, ≧83%, ≧84%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100% inhibition.

[0045] In some embodiments, the ADAMTS14 inhibitor according to the present disclosure is selected from an agent capable of reducing gene and / or protein expression of ADAMTS14 and an agent capable of binding to and inhibiting the activity of ADAMTS14. Agents capable of reducing gene and / or protein expression of ADAMTS14 and agents capable of inhibiting the activity of ADAMTS14 can be identified using the assays described above. Agents capable of binding to and inhibiting the activity of ADAMTS14 can be, for example, peptides / polypeptides, nucleic acids, or small molecules. Agents capable of binding to and inhibiting the activity of ADAMTS14 can be identified by screening libraries of such molecules for the ability to bind to and inhibit the activity of ADAMTS14.

[0046] Peptides / polypeptides capable of binding to ADAMTS14 and inhibiting its activity can be, for example, antibodies (immunoglobulins, e.g., monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific (e.g., bispecific) antibodies), antibody-derived molecules (e.g., antigen-binding fragments of antibodies (e.g., Fv, scFv, Fab, scFab, F(ab')2, Fab2, diabodies, triabodies, scFv-Fc, minibodies, single domain antibodies (e.g., VhH), etc.), peptide aptamers, thioredoxins, monobodies, anticalins, Kunitz domains, avimers, knottins, finomers, atrimers, DARPins, affibodies, nanobodies (i.e., single domain antibodies (sdAb)), affilins, armadillo repeat proteins (ArmRPs), or OBodies (see, for example, Reverdatto et al., Curr Top Med Chem. 2015; 15(12): 2015, which is hereby incorporated by reference in its entirety). 1082-1101; see also, e.g., Boersma et al., J Biol Chem (2011) 286:41273-85 and Emanuel et al., Mabs (2011) 3:38-48).

[0047] The nucleic acid capable of binding to ADAMTS14 and inhibiting its activity can be, for example, a nucleic acid aptamer. Nucleic acid aptamers can be identified and / or produced, for example, by the method of Systematic Evolution of Ligands by EXponential enrichment (SELEX), as reviewed in Zhou and Rossi, Nat Rev Drug Discov. (2017) 16(3):181-202, or by developing SOMAmers (modified aptamers with slow off-rates) (Gold L et al. (2010) PLoS ONE 5(12):e15004). Nucleic acid aptamers can include DNA and / or RNA, and can be single-stranded or double-stranded. They can include chemically modified nucleic acids, for example, sugars and / or phosphates and / or bases are chemically modified, for example, to improve the stability of the aptamer and / or to increase the resistance of the aptamer to degradation. ADAMTS14 binding aptamers include catalog number CTApt-1291 from Creative Biolabs. As used herein, "small molecule" refers to an organic compound of low molecular weight (<1000 Daltons, typically about 300-700 Daltons). Small molecule inhibitors of ADAMTS14 can be identified by screening libraries of such small molecules for their ability to bind to and inhibit the activity of ADAMTS14.

[0048] Agents that can reduce ADAMTS14 gene and / or protein expression include, for example, inhibitory nucleic acids and site-specific nuclease (SSN) systems. Inhibitory nucleic acids according to the present disclosure may comprise or consist of DNA and / or RNA. Inhibitory nucleic acids may be single-stranded (e.g., in the case of antisense oligonucleotides (e.g., gapmers)). Inhibitory nucleic acids may be double-stranded or may include double-stranded regions (e.g., in the case of siRNAs, shRNAs, etc.). Inhibitory nucleic acids may include both double-stranded and single-stranded regions (e.g., in the case of shRNA and pre-miRNA molecules that are double-stranded in the stem region of the hairpin structure and single-stranded in the loop region of the hairpin structure). In some embodiments, an inhibitory nucleic acid according to the present disclosure may be an antisense nucleic acid as described herein. In some embodiments, an inhibitory nucleic acid may comprise an antisense nucleic acid as described herein. In some embodiments, an inhibitory nucleic acid may encode an antisense nucleic acid as described herein.

[0049] As used herein, "antisense nucleic acid" refers to a nucleic acid (e.g., DNA or RNA) that is complementary to at least a portion of a target nucleotide sequence (e.g., of an RNA encoding ADAMTS14). Antisense nucleic acids according to the present disclosure are preferably single-stranded nucleic acids that bind to a target nucleotide sequence via complementary Watson-Crick base pairing. Complementary base pairing can involve hydrogen bonds between complementary base pairs. Antisense nucleic acids can be provided as single-stranded molecules, such as in the case of antisense oligonucleotides, or can be included in double-stranded molecular species, such as in the case of siRNA, shRNA, and pre-miRNA molecules. The complementary base pairing between the antisense nucleic acid and its target nucleotide sequence may be complete. In such an embodiment, the antisense nucleic acid comprises or consists of the reverse complement of its target nucleotide sequence, and complementary base pairing occurs between each nucleotide of the target nucleotide sequence and the complementary nucleotide in the antisense nucleic acid. Alternatively, the complementary base pairing between the antisense nucleic acid and its target nucleotide sequence may be incomplete / partial. In such an embodiment, complementary base pairing occurs between some but not all nucleotides of the target nucleotide sequence and the complementary nucleotide in the antisense nucleic acid.

[0050] Such binding between nucleic acids through complementary base pairing can be referred to as "hybridization." Through binding to its target nucleotide sequence, the antisense nucleic acid can form a nucleic acid complex comprising (i) the antisense nucleic acid and (ii) a target nucleic acid that comprises the target nucleotide sequence. The nucleotide sequence of the antisense nucleic acid is sufficiently complementary to its target nucleotide sequence so that it binds or hybridizes to the target nucleotide sequence. It will be understood that the antisense nucleic acid preferably has a high degree of sequence identity with the reverse complement of its target nucleotide sequence. In some embodiments, the antisense nucleic acid comprises or consists of a nucleotide sequence that has at least 75% sequence identity with the reverse complement of its target nucleotide sequence (e.g., at least one of 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity). In some embodiments, an antisense nucleic acid according to this disclosure comprises a nucleotide sequence that is the reverse complement of its target nucleotide sequence, or a nucleotide sequence that contains 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) substitutions relative to the reverse complement of its target nucleotide sequence.

[0051] In some embodiments, a target nucleotide sequence for an antisense nucleic acid according to the present disclosure comprises or consists of one of 5 to 100 nucleotides, such as 10 to 80, 12 to 50, or 15 to 30 nucleotides (e.g., 20 to 27, e.g., about 21). In some embodiments, a target nucleotide sequence for an antisense nucleic acid according to the present disclosure comprises or consists of DNA and / or RNA. In some embodiments, a target nucleotide sequence for an antisense nucleic acid according to the present disclosure comprises or consists of RNA. In some embodiments, the antisense nucleic acid reduces / prevents transcription of a nucleic acid containing its target nucleotide sequence, hi some embodiments, the antisense nucleic acid reduces / prevents the association of factors required for normal transcription (e.g., enhancers, RNA polymerase) with a nucleic acid containing its target nucleotide sequence. In some embodiments, an antisense nucleic acid increases / enhances degradation of a nucleic acid that contains its target nucleotide sequence, e.g., through RNA interference. In some embodiments, an antisense nucleic acid reduces / prevents translation of a nucleic acid that contains its target nucleotide sequence, e.g., through RNA interference or antisense degradation via RNase H activity.

[0052] RNA interference is described, for example, in Agrawal et al., Microbiol. Mol. Bio. Rev. (2003) 67(4): 657-685 and Hu et al., Sig. Transduc. Tar. Ther. (2020) 5(101), both of which are hereby incorporated by reference in their entirety. Briefly, double-stranded RNA molecules are recognized by the Argonaute component of the RNA-induced silencing complex (RISC). The double-stranded RNA is separated into single strands and integrated into active RISC by the RISC loading complex (RLC). The strands integrated into RISC bind to their target RNA through complementary base pairing, and depending on the identity of the RNA integrated into RISC and the degree of complementarity with the target RNA, RISC then cleaves the target RNA, resulting in its degradation, or otherwise blocks ribosome access, thereby preventing its translation. RNAi-based therapeutics have been approved for several indications (see, e.g., Kim, Chonnam Med J. (2020) 56(2): 87-93). In some embodiments, an antisense nucleic acid reduces / prevents normal post-transcriptional processing (e.g., splicing and / or translation) of a nucleic acid that contains its target nucleotide sequence. In some embodiments, an antisense nucleic acid reduces or alters the splicing of a pre-mRNA that contains its target nucleotide sequence into a mature mRNA. In some embodiments, an antisense nucleic acid reduces the translation of an mRNA that contains its target nucleotide sequence into a protein.

[0053] In some embodiments, the antisense nucleic acid reduces / prevents the association of factors required for normal post-transcriptional processing (e.g., components of the spliceosome) with a nucleic acid containing its target nucleotide sequence. In such cases, the antisense nucleic acid may be referred to as a "splice-switching" nucleic acid. Splice switching nucleic acids are reviewed, for example, in Haves and Hastings, Nucleic Acids Res. (2016) 44(14): 6549-6563, which is hereby incorporated by reference in its entirety. Splice switching nucleic acids include, for example, splice switching oligonucleotides (SSOs). They disrupt normal splicing of a target RNA transcript by blocking RNA:RNA base pairing and / or protein:RNA binding interactions that occur between components of the splicing machinery and the pre-mRNA. Splice switching nucleic acids can be employed to alter the number / proportion of mature mRNA transcripts that encode ADAMTS14. Splice switching nucleic acids can be designed to target specific regions of a target transcript, for example to cause skipping of an exon of interest, e.g., an exon encoding a domain / region of interest. SSOs often include modifications of the oligonucleotide sugar-phosphate backbone to reduce / prevent RNAse H degradation, such as, for example, phosphorothioate linkages, phosphorodiamidate linkages such as phosphorodiamidate morpholinos (PMOs), and may include, for example, peptide nucleic acids (PNAs), locked nucleic acids (LNAs), methoxyethyl nucleotide modifications, such as 2'O-methyl (2'OMe) and 2'-O-methoxyethyl (MOE) ribose modifications, and / or 5'-methylcytosine modifications.

[0054] In some embodiments, an antisense nucleic acid inhibits / reduces translation of a nucleic acid that contains its target nucleotide sequence, hi some embodiments, an antisense nucleic acid reduces / prevents the association of a factor required for translation (e.g., ribosomes) with a nucleic acid that contains its target nucleotide sequence. It will be understood that the target nucleotide sequence to which the antisense nucleic acid binds is a nucleotide sequence encoding a protein whose expression is desired to be inhibited. Thus, in aspects and embodiments of the present disclosure, the target nucleotide sequence for the antisense nucleic acid is the nucleotide sequence of the gene encoding ADAMTS14. In some embodiments, the target nucleotide sequence is the nucleotide sequence of the RNA encoded by the gene encoding ADAMTS14. In some embodiments, the target nucleotide sequence is the nucleotide sequence of the RNA encoding ADAMTS14. In some embodiments, the target nucleotide sequence comprises one or more nucleotides of an exon of the RNA encoding ADAMTS14. In some embodiments, the target nucleotide sequence is the nucleotide sequence of an exon of the RNA encoding ADAMTS14.

[0055] In some embodiments, the target nucleotide sequence is the nucleotide sequence of the pre-mRNA transcribed from NCBI Reference Sequence: NG_042147.1 (genomic sequence encoding human ADAMTS14). In some embodiments, the target nucleotide sequence is the nucleotide sequence of the mature mRNA sequence corresponding to NCBI Reference Sequence: NM_080722.4 (cDNA sequence encoding human ADAMTS14). In some embodiments, the target nucleotide sequence is SASI_Hs01_00087500, SASI_Hs02_00362496, SASI_Hs01_00087503, SASI_Hs02_00362497, SASI_Hs02_00362498, SASI_Hs02_00362499, SASI_Hs02_00362500, SASI_Hs02_00362501, SASI_Hs01_00087507, The target nucleotide sequence of the siRNA selected from SASI_Hs02_00362502, SASI_Hs01_00087499, SASI_Hs01_00087501, SASI_Hs01_00087502, SASI_Hs01_00087504, SASI_Hs01_00087505, SASI_Hs01_00087506, and SASI_Hs01_00087508 (Sigma-Aldrich).

[0056] In some embodiments, the target nucleotide sequence is a target nucleotide sequence of an shRNA selected from TRCN0000046703, TRCN0000046704, TRCN0000046705, TRCN0000046706, TRCN0000046707, TRCN0000415803, TRCN0000427825, and TRCN0000430560 (Sigma-Aldrich). In some embodiments, the target nucleotide sequence is the target nucleotide sequence of the siRNA of Smart Pool catalog number L-00576-00-0005 (Dharmacon). In some embodiments, the target nucleotide sequence is the target nucleotide sequence of the siRNA of siGENOME human ADAMTS14 siRNA catalog number D-005765-01 (Dharmacon). In some embodiments, the target nucleotide sequence is the target nucleotide sequence of the siRNA of siGENOME human ADAMTS14 siRNA catalog number D-005765-03 (Dharmacon).

[0057] In some embodiments, the target nucleotide sequence is a target nucleotide sequence of miR-29 (e.g., miR-29a, miR-29b, or miR-29c). In some embodiments, the target nucleotide sequence is not a target nucleotide sequence of miR-29 (e.g., miR-29a, miR-29b, or miR-29c). In some embodiments, the antisense nucleic acid comprises or consists of a sequence having at least 75% sequence identity (e.g., at least one of 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater percentage sequence identity) with the reverse complement of the nucleotide sequence of a pre-mRNA transcribed from NCBI Reference Sequence: NG_042147.1. In some embodiments, the antisense nucleic acid comprises or consists of a sequence having at least 75% sequence identity (e.g., at least one of 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater percentage sequence identity) with the reverse complement of the nucleotide sequence of the mature mRNA sequence corresponding to the NCBI reference sequence: NM_080722.4.

[0058] In some embodiments, the antisense nucleic acid is selected from the group consisting of SASI_Hs01_00087500, SASI_Hs02_00362496, SASI_Hs01_00087503, SASI_Hs02_00362497, SASI_Hs02_00362498, SASI_Hs02_00362499, SASI_Hs02_00362500, SASI_Hs02_00362501, SASI_Hs01_00087507, SASI_Hs s02_00362502, SASI_Hs01_00087499, SASI_Hs01_00087501, SASI_Hs01_00087502, SASI_Hs01_00087504, SASI_Hs01_00087505, SASI_Hs01_00087506, and SASI_Hs01_00087508 (Sigma-Aldrich). In some embodiments, the antisense nucleic acid comprises or consists of the nucleotide sequence of the guide strand of an siRNA derived from an shRNA selected from TRCN0000046703, TRCN0000046704, TRCN0000046705, TRCN0000046706, TRCN0000046707, TRCN0000415803, TRCN0000427825, and TRCN0000430560 (Sigma-Aldrich).

[0059] In some embodiments, the antisense nucleic acid comprises or consists of the nucleotide sequence of the guide strand of the siRNA Smart Pool Catalog No. L-00576-00-0005 (Dharmacon). In some embodiments, the antisense nucleic acid comprises or consists of the nucleotide sequence of the guide strand of the siRNA siGENOME Human ADAMTS14 siRNA Catalog No. D-005765-01 (Dharmacon). In some embodiments, the antisense nucleic acid comprises or consists of the nucleotide sequence of the guide strand of the siRNA siGENOME Human ADAMTS14 siRNA Catalog No. D-005765-03 (Dharmacon). In some embodiments, the antisense nucleic acid comprises or consists of the nucleotide sequence of the guide strand of miR-29 (e.g., miR-29a, miR-29b, or miR-29c). In some embodiments, the antisense nucleic acid does not comprise or consist of the nucleotide sequence of the guide strand of miR-29 (e.g., miR-29a, miR-29b, or miR-29c).

[0060] In some embodiments, the inhibitory nucleic acid is selected from siRNA, dsiRNA, miRNA, shRNA, pri-miRNA, pre-miRNA, saRNA, snoRNA, or antisense oligonucleotide (e.g., gapmer), or a nucleic acid encoding same. In some embodiments, the inhibitory nucleic acid is selected from siRNA, dsiRNA, miRNA, shRNA. In some embodiments, the inhibitory nucleic acid is an siRNA. In some embodiments, an inhibitory nucleic acid can include an antisense nucleic acid described herein, e.g., as part of a larger nucleic acid species. For example, in some embodiments, an inhibitory nucleic acid can be an siRNA, dsiRNA, miRNA, shRNA, pri-miRNA, pre-miRNA, saRNA, or snoRNA that includes an antisense nucleic acid described herein.

[0061] In some embodiments, the inhibitory nucleic acid is a small interfering RNA (siRNA). As used herein, "siRNA" refers to a double-stranded RNA molecule having a length of between 17-30 (e.g., 20-27, e.g., about 21) base pairs that can engage the RNA interference (RNAi) pathway for targeted degradation of a target RNA. A double-stranded siRNA molecule can be formed as a nucleic acid complex of RNA strands with a high degree of complementarity. In some embodiments, the siRNA molecule includes a symmetric 3' overhang (e.g., a "UU" 3' overhang), e.g., containing one or two nucleotides. The strand of the double-stranded siRNA molecule that has complementarity with the target nucleotide sequence (i.e., the antisense nucleic acid) can be referred to as the "guide" strand, and the other strand can be referred to as the "passenger" strand. The structure and function of siRNA are described, for example, in Kim and Rossi, Biotechniques. (2008) 44(5): 613-616. In some embodiments, the guide strand of a siRNA according to this disclosure may comprise or consist of an antisense nucleic acid according to the antisense nucleic acid embodiments described herein.

[0062] In some embodiments, the inhibitory nucleic acid is selected from the group consisting of SASI_Hs01_00087500, SASI_Hs02_00362496, SASI_Hs01_00087503, SASI_Hs02_00362497, SASI_Hs02_00362498, SASI_Hs02_00362499, SASI_Hs02_00362500, SASI_Hs02_00362501, SASI_Hs01_00087507, SASI SASI_Hs01_00087502, SASI_Hs01_00087499, SASI_Hs01_00087501, SASI_Hs01_00087502, SASI_Hs01_00087504, SASI_Hs01_00087505, SASI_Hs01_00087506, and SASI_Hs01_00087508 (Sigma-Aldrich). In some embodiments, the inhibitory nucleic acid comprises guide and passenger strands of an siRNA derived from an shRNA selected from TRCN0000046703, TRCN0000046704, TRCN0000046705, TRCN0000046706, TRCN0000046707, TRCN0000415803, TRCN0000427825, and TRCN0000430560 (Sigma-Aldrich).

[0063] In some embodiments, the inhibitory nucleic acid comprises the guide and passenger strands of siRNA Smart Pool Catalog No. L-00576-00-0005 (Dharmacon). In some embodiments, the inhibitory nucleic acid comprises the guide and passenger strands of siRNA siGENOME Human ADAMTS14 siRNA Catalog No. D-005765-01 (Dharmacon). In some embodiments, the inhibitory nucleic acid comprises the guide and passenger strands of siRNA siGENOME Human ADAMTS14 siRNA Catalog No. D-005765-03 (Dharmacon). In some embodiments, the inhibitory nucleic acid comprises a guide and passenger strand of miR-29 (e.g., miR-29a, miR-29b, or miR-29c). In some embodiments, the inhibitory nucleic acid does not comprise a guide and passenger strand of miR-29 (e.g., miR-29a, miR-29b, or miR-29c).

[0064] In some embodiments, the inhibitory nucleic acid is selected from the group consisting of SASI_Hs01_00087500, SASI_Hs02_00362496, SASI_Hs01_00087503, SASI_Hs02_00362497, SASI_Hs02_00362498, SASI_Hs02_00362499, SASI_Hs02_00362500, SASI_Hs02_00362501, SASI_Hs01_000875 07, SASI_Hs02_00362502, SASI_Hs01_00087499, SASI_Hs01_00087501, SASI_Hs01_00087502, SASI_Hs01_00087504, SASI_Hs01_00087505, SASI_Hs01_00087506, and SASI_Hs01_00087508 (Sigma-Aldrich). In some embodiments, the inhibitory nucleic acid is an shRNA selected from TRCN0000046703, TRCN0000046704, TRCN0000046705, TRCN0000046706, TRCN0000046707, TRCN0000415803, TRCN0000427825, and TRCN0000430560 (Sigma-Aldrich).

[0065] In some embodiments, the inhibitory nucleic acid is an siRNA of Smart Pool catalog number L-00576-00-0005 (Dharmacon). In some embodiments, the inhibitory nucleic acid is an siRNA of siGENOME human ADAMTS14 siRNA catalog number D-005765-01 (Dharmacon). In some embodiments, the inhibitory nucleic acid is an siRNA of siGENOME human ADAMTS14 siRNA catalog number D-005765-03 (Dharmacon). In some embodiments, the inhibitory nucleic acid is miR-29 (e.g., miR-29a, miR-29b, or miR-29c). In some embodiments, the inhibitory nucleic acid is not miR-29 (e.g., miR-29a, miR-29b, or miR-29c). In some embodiments, the inhibitory nucleic acid is a Dicer small interfering RNA (dsiRNA). As used herein, "dsiRNA" refers to a double-stranded RNA molecule having a length of about 27 base pairs that is processed by Dicer into siRNA for RNAi-mediated degradation of target RNA. DsiRNA is described, for example, in Raja et al., Asian J Pharm Sci. (2019) 14(5): 497-510, which is hereby incorporated by reference in its entirety. DsiRNA can be optimized for Dicer processing and have increased potency compared to 21-mer siRNA (see, for example, Kim et al., Nat Biotechnol. (2005) 23(2): 222-226), which may relate to the link between Dicer-mediated nuclease activity and RISC loading.

[0066] In some embodiments, the inhibitory nucleic acid is a microRNA (miRNA) or a precursor thereof (e.g., pri-miRNA or pre-miRNA). miRNA molecules have a similar structure to siRNA molecules, but are endogenously encoded and derived from the processing of short hairpin RNA molecules. They are initially expressed as long primary transcripts (pri-miRNAs), which are processed in the nucleus to a 60-70 nucleotide hairpin (pre-miRNA), which is further processed in the cytoplasm to smaller species that interact with RISC and the target mRNA. miRNAs contain a "seed sequence" that is essential for binding to the target mRNA. The seed sequence usually contains 6 nucleotides and is located at positions 2-7 at the 5' end of the miRNA.

[0067] Cushing et al., miRNA Biochem Cell Biol. (2015) 93(2):109-18, a review article on a miRNA designated miR-29, discloses that miR-29 is an inhibitor of expression of a wide range of genes involved in extracellular matrix production, cross-linking, and degradation, genes involved in basement membrane function, PDGF genes, and interleukin and TGF genes (see, e.g., FIG. 1). ADAMTS14 is identified as a gene whose expression is repressed by miR-29. In some embodiments, the ADAMTS14 inhibitor according to the present disclosure is miR-29 (e.g., miR-29a, miR-29b, or miR-29c). In some embodiments, the ADAMTS14 inhibitor according to the present disclosure is not miR-29 (e.g., miR-29a, miR-29b, or miR-29c); that is, in some embodiments, the ADAMTS14 inhibitor according to the present disclosure is an ADAMTS14 inhibitor other than miR-29 (e.g., miR-29a, miR-29b, or miR-29c).

[0068] In some embodiments, the inhibitory nucleic acid is a short hairpin RNA (shRNA). The shRNA molecule comprises a sequence of nucleotides with a high degree of complementarity that associate with each other through complementary base pairing to form the stem region of the hairpin. The sequence of nucleotides with a high degree of complementarity may be linked by one or more nucleotides that form the loop region of the hairpin. The shRNA molecule may be processed (e.g., via catalytic cleavage by Dicer) to form an siRNA or miRNA molecule. The shRNA molecule may have a length of between 35-100 (e.g., 40-70) nucleotides. The stem region of the hairpin may have a length of between 17-30 (e.g., 20-27, e.g., about 21) base pairs. The stem region may include GU pairings that stabilize the hairpin structure. siRNA, dsiRNA, miRNA, and shRNA for targeted inhibition of ADAMTS14 gene and / or protein expression can be identified / designed according to principles and / or using tools well known to those skilled in the art. Parameters and tools for designing siRNA and shRNA molecules are described, for example, in Fakhr et al., Cancer Gene Therapy (2016) 23:73-82 (hereby incorporated by reference in its entirety). Software that can be used by those skilled in the art to design such molecules is summarized in Table 1 of Fakhr et al., Cancer Gene Therapy (2016) 23:73-82, and includes, for example, siRNA Wizard (InvivoGen). Details for designing and producing such molecules can be found on the websites of commercial vendors such as Ambion, Dharmacon, GenScript, Invitrogen, and OligoEngine.

[0069] In some embodiments, the inhibitory nucleic acid is an antisense oligonucleotide (ASO). An ASO is a single-stranded nucleic acid molecule that comprises or consists of an antisense nucleic acid against a target nucleotide sequence. The antisense oligonucleotide according to the present disclosure can comprise or consist of the antisense nucleic acid described herein. ASOs may modify the expression of RNA molecules containing their target nucleotide sequences by altering splicing or by directing RNase H to degrade RNA containing the target nucleotide sequence. RNase H recognizes a nucleic acid complex molecule formed when an ASO binds to an RNA containing its target nucleotide sequence. An ASO according to the present disclosure may comprise or consist of an antisense nucleic acid according to the present disclosure. An ASO may comprise 17-30 (e.g., 20-27, e.g., about 21) nucleotides in length. Many ASOs are designed as chimeras that contain a mix of bases with different chemical properties, or as gapmers that contain a central DNA portion surrounded by "wings" of modified nucleotides. ASOs are described, for example, in Scoles et al., Neurol Genet. 2019 Apr; 5(2): e323. ASOs may also contain modifications of the sugar-phosphate backbone to increase their stability and / or reduce / prevent RNAse H degradation, such as, for example, phosphorothioate linkages, phosphorodiamidate linkages such as phosphorodiamidate morpholinos (PMOs), and may contain, for example, peptide nucleic acids (PNAs), locked nucleic acids (LNAs), methoxyethyl nucleotide modifications, such as 2'O-methyl (2'OMe) and 2'-O-methoxyethyl (MOE) ribose modifications, and / or 5'-methylcytosine modifications.

[0070] Inhibitory nucleic acids according to the present disclosure may include chemically modified nucleotides, e.g., phosphonates and / or riboses and / or bases are chemically modified. Such modifications may affect the activity, specificity, and / or stability of the nucleic acid. One or more (e.g., one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or all) nucleotides of the inhibitory nucleic acid may include such chemical modifications. Modifications contemplated according to the inhibitory nucleic acids of the present disclosure include those described in Hu et al., Sig. Transduc. Tar. Ther. (2020) 5(101), which is incorporated by reference above, particularly those shown in Figure 2 of Hu et al., Sig. Transduc. Tar. Ther. (2020) 5(101). Further modifications contemplated according to the inhibitory nucleic acids according to the present disclosure include those described in Selvam et al., Chem Biol Drug Des. (2017) 90(5): 665-678, which is hereby incorporated by reference in its entirety.

[0071] In some embodiments, the inhibitory nucleic acid according to the present disclosure comprises one or more nucleotides that include a phosphonate modification. In some embodiments, the phosphonate modification can be selected from phosphorothioate (e.g., Rp isomer, Sp isomer), phosphorodithioate, methyl phosphonate, methoxypropyl phosphonate, 5'-(E)-vinyl phosphonate, 5'-methyl phosphonate, (S)-5'-C-methyl with phosphate, 5'-phosphorothioate, and peptide nucleic acid. In some embodiments, the inhibitory nucleic acid comprises one or more nucleotides that include a phosphorothioate modification. In some embodiments, the inhibitory nucleic acid according to the present disclosure comprises one or more nucleotides comprising a ribose modification. In some embodiments, the ribose modification may be selected from 2'-O-methyl, 2'-O-methoxyethyl, 2'-fluoro, 2'-deoxy-2'-fluoro, 2'-methoxyethyl, 2'-O-alkyl, 2'-O-allyl, 2'-C-allyl, 2'-deoxy, 2'-hydroxyl, 2'-arabino-fluoro, 2'-O-benzyl, 2'-O-methyl-4-pyridine, locked nucleic acid, (S)-cEt-BNA, tricyclo-DNA, PMO, unlocked nucleic acid, hexitol nucleic acid, and glycol nucleic acid. In some embodiments, the inhibitory nucleic acid comprises one or more nucleotides comprising a 2'-O-methyl modification. In some embodiments, the inhibitory nucleic acid comprises one or more nucleotides comprising a 2'-fluoro modification.

[0072] In some embodiments, an inhibitory nucleic acid according to the present disclosure comprises one or more nucleotides that include a base modification. In some embodiments, the base modification may be selected from pseudouridine, 2'-thiouridine, N6'-methyladenosine, 5'-methylcytidine, 5'-fluoro-2'-deoxyuridine, N-ethylpiperidine 7'-EAA triazole modified adenine, N-ethylpiperidine 6'-triazole modified adenine, 6'-phenylpyrrolo-cytosine, 2',4'-difluorotoluyl ribonucleoside, and 5'-nitroindole. In some embodiments, an inhibitory nucleic acid according to the present disclosure comprises one or more nucleotides that include a phosphorothioate modification, one or more nucleotides that include a 2'-O-methyl modification, and one or more nucleotides that include a 2'-fluoro modification. In embodiments in which the inhibitory nucleic acid includes nucleotides that include chemical modifications as described herein, the nucleotide sequence is nevertheless evaluated for purposes of sequence comparison according to the present disclosure as if the equivalent unmodified nucleotides were present instead.

[0073] In some embodiments, an inhibitory nucleic acid (e.g., an siRNA) according to the disclosure has an IC of ≦1 μM, e.g., one of ≦500 nM, ≦100 nM, ≦75 nM, ≦50 nM, ≦40 nM, ≦30 nM, ≦20 nM, ≦15 nM, ≦12.5 nM, ≦10 nM, ≦9 nM, ≦8 nM, ≦7 nM, ≦6 nM, ≦5 nM, ≦4 nM ≦3 nM, ≦2 nM, ≦1 nM, ≦900 pM, ≦800 pM, ≦700 pM, ≦600 pM, ≦500 pM, ≦400 pM, ≦300 pM, ≦200 pM, ≦100 pM, ≦50 pM, ≦40 pM, ≦30 pM, ≦20 pM, ≦10 pM, or ≦1 pM. 50 and can inhibit ADAMTS14 gene and / or protein expression. In some embodiments, an inhibitory nucleic acid (e.g., siRNA) according to the disclosure has an IC of ≦1 nM, ≦900 pM, ≦800 pM, ≦700 pM, ≦600 pM, ≦500 pM, ≦400 pM, ≦300 pM, ≦200 pM, ≦100 pM, ≦50 pM, ≦40 pM, ≦30 pM, ≦20 pM, ≦10 pM, or ≦1 pM. 50 and inhibiting ADAMTS14 gene expression (e.g., as determined by qRT-PCR).

[0074] In some embodiments, an inhibitory nucleic acid (e.g., siRNA) according to the disclosure has an IC of ≦1 nM, ≦900 pM, ≦800 pM, ≦700 pM, ≦600 pM, ≦500 pM, ≦400 pM, ≦300 pM, ≦200 pM, ≦100 pM, ≦50 pM, ≦40 pM, ≦30 pM, ≦20 pM, ≦10 pM, or ≦1 pM. 50 and inhibiting the protein expression of ADAMTS14 (eg, as determined by ELISA). A site-specific nuclease (SSN) system may be employed to reduce gene and / or protein expression of ADAMTS14. According to such an embodiment, reduction of gene and / or protein expression may be achieved by employing the SSN system to modify the nucleic acid encoding ADAMTS14. The SSN system may be capable of modifying the gene encoding ADAMTS14. The SSN system may be capable of introducing insertions, substitutions, or deletions into the nucleic acid sequence encoding ADAMTS14. Modification of the nucleic acid sequence encoding ADAMTS14 by the SSN system may reduce or prevent expression of a polypeptide according to SEQ ID NO: 1, 2, 3, or 4 from the modified nucleic acid sequence. Modification of the nucleic acid sequence encoding ADAMTS14 by the SSN system in a cell may have the result that the cell lacks a nucleic acid encoding a polypeptide according to SEQ ID NO: 1, 2, 3, or 4.

[0075] The SSN system may be capable of introducing a premature stop codon into a sequence transcribed from ADAMTS 14. Modification of a nucleic acid sequence encoding ADAMTS 14 by the SSN system may have the result that the modified nucleic acid encodes a truncated and / or non-functional ADAMTS 14 polypeptide, and / or an ADAMTS 14 polypeptide that is misfolded and / or degraded. Gene editing using SSN is described, for example, in Eid and Mahfouz, Exp Mol Med. 2016 Oct; 48(10): e265, which is hereby incorporated by reference in its entirety. An enzyme capable of creating a site-specific double-strand break (DSB) can be engineered to introduce a DSB into a target nucleic acid sequence of interest. A DSB can be repaired by error-prone non-homologous end joining (NHEJ), in which the two ends of the break are rejoined, often with the insertion or deletion of nucleotides. Alternatively, a DSB can be repaired by advanced homology-directed repair (HDR), in which a DNA template with ends homologous to the break site is provided and introduced at the site of the DSB. SSNs that can be engineered to generate target nucleic acid sequence-specific DSBs include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the clustered regularly spaced palindromic repeats / CRISPR-associated 9 (CRISPR / Cas9) system.

[0076] The ZFN system is reviewed, for example, in Umov et al., Nat Rev Genet. (2010) 11(9):636-46, which is hereby incorporated by reference in its entirety. ZFNs contain a programmable zinc finger DNA binding domain and a DNA cleavage domain (e.g., a FokI endonuclease domain). The DNA binding domain can be identified by screening zinc finger arrays that can bind to a target nucleic acid sequence. The TALEN system is reviewed, for example, in Mahfouz et al., Plant Biotechnol J. (2014) 12(8):1006-14, which is hereby incorporated by reference in its entirety. TALENs contain a programmable DNA-binding TALE domain and a DNA-cleavage domain (e.g., a FokI endonuclease domain). TALEs contain a repeat domain consisting of repeats of 33-39 amino acids that are identical except for two residues at positions 12 and 13 of each repeat, the repeat variable dipeptides (RVDs). Each RVD determines the binding of the repeat to a nucleotide in a target DNA sequence according to the following relationship: "HD" binds to C, "NI" binds to A, "NG" binds to T, and "NN" or "NK" binds to G (Moscou and Bogdanove, Science (2009) 326(5959):1501.).

[0077] CRISPR / Cas9 and related systems, such as CRISPR / Cpf1, CRISPR / C2c1, CRISPR / C2c2, and CRISPR / C2c3, are reviewed, for example, in Nakade et al., Bioengineered (2017) 8(3):265-273, which is hereby incorporated by reference in its entirety. These systems include an endonuclease (e.g., Cas9, Cpf1, etc.) and a single guide RNA (sgRNA) molecule. The sgRNA can be engineered to target the endonuclease activity to a nucleic acid sequence of interest. In some embodiments, the SSN system for reducing expression of ADAMTS14 according to the present disclosure is selected from a ZFN system, a TALEN system, a CRISPR / Cas9 system, a CRISPR / Cpf1 system, a CRISPR / C2c1 system, a CRISPR / C2c2 system, and a CRISPR / C2c3 system. For example, the SSN system may employ a nucleic acid encoding a CRISPR / Cas9 system. The nucleic acid may encode a CRISPR RNA (crRNA) that targets an exon of ADAMTS14, and a transactivating crRNA (tracrRNA) for processing the crRNA into its mature form.

[0078] The ADAMTS14 inhibitor according to the present disclosure may be provided in the form of a nucleic acid that contains / encodes the ADAMTS14 inhibitor. For example, a peptide / polypeptide that can bind to ADAMTS14 and inhibit its activity, and an SSN system for reducing the expression of ADAMTS14 may be provided in the form of a nucleic acid that encodes it. Similarly, a nucleic acid that can bind to ADAMTS14 and inhibit its activity, and an inhibitory nucleic acid that can reduce the expression of ADAMTS14 may be provided in the form of a nucleic acid that contains / encodes it. The nucleic acid comprising / encoding an ADAMTS14 inhibitor according to the present disclosure may comprise or consist of DNA and / or RNA. In some embodiments, the nucleic acid comprising / encoding an ADAMTS14 inhibitor according to the present disclosure may be contained in a vector. The vector may facilitate the delivery of the nucleic acid comprising / encoding an ADAMTS14 inhibitor. The vector may be an expression vector that includes the elements required to express the nucleic acid comprising / encoding an ADAMTS14 inhibitor.

[0079] As used herein, a "vector" is a nucleic acid molecule used as a vehicle to transfer exogenous nucleic acid into a cell. A vector may be a vector for expression of a nucleic acid in a cell. Such a vector may include a promoter sequence operably linked to a nucleotide sequence encoding a sequence to be expressed. The vector may also include a termination codon and an expression enhancer. Any suitable vector, promoter, enhancer, and termination codon known in the art may be used to express a nucleic acid from a vector according to the present disclosure. The term "operably linked" can include the situation where a selected nucleic acid sequence and a regulatory nucleic acid sequence (e.g., a promoter and / or enhancer) are covalently linked in such a manner that expression of the nucleic acid sequence is under the influence or control of the regulatory sequence (thereby forming an expression cassette). Thus, a regulatory sequence is operably linked to a selected nucleic acid sequence if the regulatory sequence is capable of affecting the transcription of the nucleic acid sequence.

[0080] Suitable vectors include plasmids, binary vectors, DNA vectors, mRNA vectors, viral vectors (e.g., gamma retroviral vectors (e.g., murine leukemia virus (MLV)-derived vectors), lentiviral vectors, adenoviral vectors, adeno-associated viral vectors, lentiviral vectors, vaccinia viral vectors, and herpes viral vectors), transposon-based vectors, and artificial chromosomes (e.g., yeast artificial chromosomes), as described, for example, in Maus et al., Annu Rev Immunol (2014) 32:189-225 or Morgan and Boyerinas, Biomedicines 2016 4, 9, both of which are hereby incorporated by reference in their entirety. In a preferred embodiment, the vector is an adeno-associated viral (AAV) vector or a lentiviral vector. In some embodiments, the vector may be a eukaryotic vector, e.g., a vector that contains elements necessary for expression of a nucleic acid from the vector in a eukaryotic cell. In some embodiments, the vector may be a mammalian vector, e.g., containing a cytomegalovirus (CMV) or SV40 promoter driving expression. In some embodiments, the vector includes a cell or tissue specific promoter. In some embodiments, the vector includes a fibroblast specific promoter.

[0081] In some embodiments, vectors are selected based on the tropism of the cell type / tissue / organ that is desired to deliver nucleic acid.In some embodiments, vectors are selected based on the tropism of the cell type / tissue / organ that is desired to express ADAMTS14 inhibitor.For example, it may be desirable to deliver nucleic acid / express ADAMTS14 inhibitor in the cell type / tissue / organ that is affected by the disease that is treated / prevented according to the present disclosure (e.g., the cell type / tissue / organ that shows symptoms of the disease). In a preferred embodiment, the vector is an adeno-associated virus vector. Adeno-associated virus vectors and their use for vector gene therapy are reviewed, for example, in Wang et al., Nat. Rev. Drug Discov. (2019) 18: 358-378 and Li and Samulski, Nat. Rev. Genet. (2020) 12: 255-272, both of which are hereby incorporated by reference in their entirety. In some embodiments, the vector can be an adeno-associated virus vector as described in Wang et al., Nat. Rev. Drug Discov. (2019) 18: 358-378. In some embodiments, the vector can be an adeno-associated virus vector as described in Li and Samulski, Nat. Rev. Genet. (2020) 12: 255-272. A peptide / polypeptide capable of binding to ADAMTS14 and inhibiting its activity in accordance with the present disclosure can be produced intracellularly by transcription from a vector encoding the peptide / polypeptide and subsequent translation of the transcribed RNA.

[0082] The nucleic acid aptamer that can bind to ADAMTS14 and inhibit its activity can be produced in a cell by transcription from a vector that codes for the nucleic acid aptamer. The shRNA molecule according to the present disclosure can be produced in a cell by transcription from a vector that codes for the shRNA. The shRNA can be produced in a cell by transfecting a vector that codes for a shRNA sequence under the control of an RNA polymerase promoter into a cell. The siRNA molecule according to the present disclosure can be produced in a cell by transcription from a vector that codes for a shRNA that codes for / contains the siRNA, and then processing the shRNA molecule by intracellular Dicer to form a siRNA molecule. An ADAMTS14 inhibitor according to the present disclosure may be provided in the form of a cell comprising a nucleic acid comprising / encoding an ADAMTS14 inhibitor, which may be contained, for example, in a vector as described above. The items of the disclosure (i.e., ADAMTS14 inhibitors, nucleic acids comprising / encoding ADAMTS14 inhibitors, vectors comprising such nucleic acids, cells comprising such nucleic acids or vectors) may be formulated as compositions suitable for clinical use (i.e., as medicaments or pharmaceutical compositions).

[0083] Such compositions may contain one or more pharma- ceutically acceptable components well known to those skilled in the art, including, but not limited to, pharma- ceutically acceptable carriers, adjuvants, excipients, diluents, fillers, buffers, preservatives, antioxidants, lubricants, stabilizers, solubilizers, surfactants (e.g., wetting agents), masking agents, colorants, flavoring agents, and sweetening agents. The term "pharmacologically acceptable" as used herein refers to compounds, ingredients, materials, compositions, dosage forms, etc., that are within the scope of sound medical judgment, suitable for use in contact with the tissues of the subject (e.g., human) in question, without undue toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio. Each carrier, adjuvant, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation. Suitable carriers, adjuvants, excipients, etc. can be found in standard pharmaceutical texts, such as Remington's 'The Science and Practice of Pharmacy' (ed. A. Adejare), 23rd Edition (2020), Academic Press. The composition may be prepared for local, parenteral, systemic, intracavitary, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intravitreal, intraconjunctival, subretinal, suprachoroidal, subcutaneous, intradermal, intrathecal, oral, nasal, or transdermal routes of administration, which may include injection or infusion. A suitable formulation may include the selected agent in a sterile or isotonic medium. The formulation and mode of administration may be selected according to the agent to be administered and the disease to be treated / prevented.

[0084] Nucleic acids (including inhibitory nucleic acids, expression vectors), cells, and compositions in accordance with the present disclosure may be modified and / or formulated to facilitate delivery to and / or uptake by cells / tissues of interest, e.g., fibroblasts. Strategies for targeted delivery of such species are reviewed, for example, in Li et al., Int. J. Mol. Sci. (2015) 16: 19518-19536 and Fu et al., Bioconjug Chem. (2014) 25(9): 1602-1608, which are hereby incorporated by reference in their entireties. In particular, nucleic acids according to the present disclosure may employ delivery platforms as described in Hu et al., Sig. Transduc. Tar. Ther. (2020) 5(101), incorporated by reference above, or Tatiparti et al. 'siRNA Delivery Strategies: A Comprehensive Review of Recent Developments.' Ed. Thomas Nann. Nanomaterials 7.4 (2017): 77, and Lehto T et al., Adv Drug Deliv Rev. 2016, 106(Pt A):172-182, which are hereby incorporated by reference in their entireties.

[0085] In some embodiments, the articles of the present disclosure may be encapsulated in nanoparticles or liposomes. In some embodiments, the articles of the present disclosure may be associated (covalently or non-covalently) with cell penetrating peptides (e.g., protein transduction domains, trochoid peptides, arginine-rich peptides, vectocell peptides), cationic polymers, cationic lipids, or viral carriers. The nanoparticles can be organic, such as micelles, liposomes, proteins, solid lipid particles, solid polymer particles, dendrimers, and polymeric therapeutics. The nanoparticles can be inorganic, such as nanotubes or metal particles, optionally with added organic molecules. In some embodiments, the nanoparticles are those described in Chen et al., Mol Ther Methods Clin Dev. (2016) 3:16023, which is hereby incorporated by reference in its entirety. In some embodiments, the nanoparticles are PLGA, polypeptides, poly(β-amino esters), DOPE, β-cyclodextrin-containing polycations, linear PEI, PAMAM dendrimers, branched PEI, chitosan, or polyphosophoester nanoparticles.

[0086] In some embodiments, a nucleic acid according to the present disclosure includes a modification that incorporates one or more moieties that facilitate delivery to and / or uptake by a cell type or tissue of interest. In some embodiments, a nucleic acid according to the present disclosure is linked (e.g., chemically conjugated) to one or more moieties that facilitate delivery to and / or uptake by a cell type or tissue of interest. Modifications and formulations of nucleic acids to facilitate targeted delivery to a cell type and / or tissue of interest are described, for example, in Lorenzer et al., J Control Release (2015) 203:1-15, which is hereby incorporated by reference in its entirety. A moiety that facilitates delivery to and / or uptake by a cell type or tissue of interest may selectively bind to the target cell type / tissue of interest. The moiety may facilitate crossing of the cell membrane of the cells of the target cell type and / or tissue of interest. The moiety may bind to a molecule expressed on the cell surface of the target cell type / tissue of interest. The moiety may facilitate internalization (e.g., by endocytosis) of the nucleic acid by the target cell type / tissue of interest. Moieties that facilitate delivery to and / or uptake by a cell type or tissue of interest are described, for example, in Benizri et al., Bioconjug Chem. (2019) 30(2): 366-383, which is hereby incorporated by reference in its entirety. Such moieties include, for example, N-acetylgalactosamine (GalNAc), α-tocopherol, cell penetrating peptides, nucleic acid aptamers, antibodies and antigen-binding fragments / derivatives thereof, cholesterol, squalene, polyethylene glycol (PEG), fatty acids (e.g., palmitic acid), and nucleolipid moieties.

[0087] Inhibition of fibrosis-induced fibroblasts Aspects and embodiments of the present disclosure relate to the use of ADAMTS14 inhibitors to inhibit the generation of and / or processes mediated by pro-fibrotic fibroblasts (eg, myofibroblasts). Fibroblasts are the most common cell type found in connective tissues. The development and biology of fibroblasts is described, for example, in Plikus et al. Cell (2021) 184(15):3852-3872 (hereby incorporated by reference in its entirety), and the role of fibroblasts in fibrosis is described, for example, in Kendall and Feghali-Bostwick, Front. Pharmacol. (2014) 5:123 (hereby incorporated by reference in its entirety). As referred to herein, a "pro-fibrotic fibroblast" is a fibroblast that actively contributes to tissue / organ fibrosis through the production (i.e., secretion, deposition) of extracellular matrix (ECM) components (e.g., collagen).

[0088] Profibrotic fibroblasts include myofibroblasts, which are described, for example, in Baum and Duffy, J Cardiovasc Pharmacol. (2011) 57(4): 376-379 and Bagalad et al., J Oral Maxillofac Pathol. (2017) 21(3): 462-463, both of which are hereby incorporated by reference in their entirety. Myofibroblasts are contractile cells that have stress fibers that contain alpha smooth muscle actin (αSMA). Myofibroblasts can be further characterized by the expression of collagen types 1, 2, and 3, fibronectin, and / or MMP-9. Myofibroblasts can also be characterized by the production of extracellular matrix, including fibronectin (e.g., the EDA isoform of fibronectin), collagen (e.g., collagen type 1, e.g., α1 collagen type 1 and α2 collagen type 1), and hyaluronic acid. Additional markers of myofibroblasts include vimentin and palladin.

[0089] Myofibroblasts can be derived from a wide variety of different cell types. They can arise by differentiation from tissue-resident fibroblasts in response to proinflammatory mediators such as TGFβ1, IL-4, IL-6, IL-13, ET-1, IGF-II, and PDGF, and undergo a fibroblast-to-myofibroblast transition (FMT). The same factors can also activate stellate cells, such as hepatic stellate cells (HSCs) in the liver and pancreatic stellate cells in the pancreas, to develop into myofibroblasts. Epithelial cells are stimulated to undergo epithelial-mesenchymal cell transition to myofibroblasts in response to proinflammatory factors such as TGFβ1, TNFα, IL-1β, and IL-4 (see, e.g., Li et al., Exp Biol Med (Maywood). (2016) 241(1): 1-13), and TGFβ1, TNFα, and IL-1β can also induce endothelial cells to undergo endothelial-mesenchymal cell transition to myofibroblasts. Myofibroblasts also differentiate from bone marrow-derived fibrocyte precursors in response to factors including CCL21, TGFβ1, IL-4, IL-13, PDGF, and CXCL12, and proinflammatory mediators can also include smooth muscle cells that "dedifferentiate" into myofibroblasts. Myofibroblasts are the main effectors in fibrosis. Fibrosis is a form of pathological tissue remodeling characterized by the formation of excessive connective tissue as a result of excessive deposition of extracellular matrix (ECM) components (including collagen). "Excessive connective tissue" refers to an amount of connective tissue in a given location (e.g., a given tissue / organ, or a part of a given tissue / organ) that is greater than the amount of connective tissue present in that location under normal, non-pathological conditions. Similarly, "excessive deposition of ECM components" refers to a level of deposition of one or more ECM components that is greater than the level of deposition under normal, non-pathological conditions.

[0090] The cellular and molecular mechanisms of fibrosis are described in Wynn, J. Pathol. (2008) 214(2): 199-210, and Wynn and Ramalingam, Nature Medicine (2012) 18:1028-1040, both of which are hereby incorporated by reference in their entireties. Damage to tissues can occur due to a variety of stimuli, including infection, autoimmune reactions, toxins, radiation, and mechanical injury. Repair typically involves the replacement of injured cells with the same type of cells and the replacement of normal parenchymal tissue with connective tissue. Repair processes can become pathological when they are not properly controlled, resulting in excessive deposition of ECM components, with normal parenchymal tissue being replaced by connective tissue. In diseases such as idiopathic pulmonary fibrosis, liver cirrhosis, cardiovascular fibrosis, systemic sclerosis, and nephritis, extensive tissue remodeling and fibrosis can ultimately lead to organ failure and death. The main cellular effectors of fibrosis are myofibroblasts. In response to tissue injury, damaged cells and leukocytes produce pro-fibroinflammafory factors such as TGFβ, IL-13, and PDGF, which activate fibroblasts (and other myofibroblast precursors) to become αSMA-expressing myofibroblasts and guide them to the site of injury. Myofibroblasts produce large amounts of extracellular matrix components such as collagen and periostin for wound contracture and closure, as well as proinflammatory cytokines such as IL-6, and tissue remodeling factors such as MMP2 and TIMP1. Persistent / chronic infection and / or inflammation can lead to the generation of too many myofibroblasts and, consequently, the overproduction of extracellular matrix, resulting in fibrosis. In many diseases and conditions characterized by fibrosis, persistent inflammatory triggers are essential for upregulating the production of growth factors, proteolytic enzymes, angiogenic factors, and fibrogenic cytokines, which stimulate the deposition of progressively remodeling connective tissue elements and disrupt normal tissue architecture.

[0091] It has been demonstrated in the experimental examples of the present disclosure that ADAMTS14 inhibitors inhibit the expression of profibrotic genes in fibroblasts in response to stimulation with profibrotic factors (e.g., TGFβ1), see Figures 1 and 6, and Examples 2.1 and 2.6. Inhibition of ADAMTS14 has also been shown to inhibit the formation of αSMA-containing fibers in and collagen I deposition by fibroblasts in response to stimulation with such profibrotic factors, see Figure 5 and Example 2.5. Thus, the experimental examples of the present disclosure demonstrate that ADAMTS14 inhibitors are useful for inhibiting the generation of myofibroblasts and for inhibiting processes mediated by myofibroblasts. TGFβ1-mediated expression of profibrotic genes has been shown to be a conserved mechanism driving fibrosis across a wide range of tissues, and myofibroblasts have likewise been shown to be central effectors of fibrosis in a variety of tissues and in a variety of different fibrotic disorders (see, e.g., Kim et al., Cold Spring Harb Perspect Biol. (2018) 10(4):a022293). Thus, the demonstration in this example that ADAMTS14 inhibitors antagonize TGFβ1-mediated upregulation of profibrotic gene expression and inhibit myofibroblast generation and profibrotic activity indicates that ADAMTS14 inhibitors are generally useful for treating / preventing fibrosis across all tissues and a wide range of fibrotic disorders.

[0092] Thus, the present disclosure provides an ADAMTS14 inhibitor for use in a method comprising inhibiting the generation of myofibroblasts from myofibroblast precursor cells.Also provided is the use of an ADAMTS14 inhibitor in a method for inhibiting the generation of myofibroblasts from myofibroblast precursor cells.Also provided is a method for inhibiting the generation of myofibroblasts from myofibroblast precursor cells, comprising contacting myofibroblast precursor cells with an ADAMTS14 inhibitor.In some embodiments, the myofibroblast precursor cells can be selected from fibroblasts, stellate cells (e.g., hepatic stellate cells, pancreatic stellate cells), epithelial cells, endothelial cells, fibrocytes, and smooth muscle cells. In some embodiments, the myofibroblast precursor cells are fibroblasts. In some embodiments, the uses and methods include inhibiting fibroblast to myofibroblast transition (FMT). The present disclosure also provides an ADAMTS14 inhibitor for use in a method comprising inhibiting TGFβ-mediated signal transduction. Also provided is the use of an ADAMTS14 inhibitor in a method for inhibiting TGFβ-mediated signal transduction. Also provided is a method for inhibiting TGFβ-mediated signal transduction, comprising contacting a cell (e.g., a myofibroblast or a myofibroblast precursor cell) with an ADAMTS14 inhibitor.

[0093] In some embodiments, the uses and methods include inhibition of TGFβ-mediated signaling in myofibroblasts and / or myofibroblast precursor cells (eg, fibroblasts). The present disclosure also provides an ADAMTS14 inhibitor for use in a method comprising inhibiting a process mediated by myofibroblasts.The use of an ADAMTS14 inhibitor in a method for inhibiting a process mediated by myofibroblasts is also provided.A method for inhibiting a process mediated by myofibroblasts is also provided, comprising contacting myofibroblasts or myofibroblast precursor cells with an ADAMTS14 inhibitor.In some embodiments, the process mediated by myofibroblasts can be selected from collagen (e.g., type I collagen) production / deposition, extracellular matrix production / deposition, fibrosis, and tissue remodeling (e.g., epithelial tissue remodeling). According to such aspects and embodiments, myofibroblasts and / or myofibroblast precursor cells may be contacted with an ADAMTS14 inhibitor in vitro or in vivo. In some embodiments, the ADAMTS14 inhibitor is introduced into the myofibroblasts and / or myofibroblast precursor cells (e.g., by transfection or transduction).

[0094] In some embodiments, an ADAMTS14 inhibitor according to the present disclosure reduces the number / proportion of myofibroblasts in a given assay by less than 1-fold, e.g., ≦0.99-fold, ≦0.95-fold, ≦0.9-fold, ≦0.85-fold, ≦0.8-fold, ≦0.75-fold, ≦0.7-fold, ≦0.65-fold, ≦0.6-fold, ≦0.55-fold, ≦0.5-fold, ≦0.45-fold, ≦0.4-fold, ≦0.35-fold, ≦0.3-fold, ≦0.25-fold, ≦0.2-fold, ≦0.15-fold, ≦0.1-fold, ≦0.05-fold, or ≦0.01-fold, the number / proportion of such cells observed in the absence of the ADAMTS14 inhibitor or in the presence of the same amount of a control agent known to have no such inhibitory activity. In some embodiments, an ADAMTS14 inhibitor according to the present disclosure reduces the number / proportion of myofibroblasts in a given assay to less than 100%, e.g., one of: ≦99%, ≦95%, ≦90%, ≦85%, ≦80%, ≦75%, ≦70%, ≦65%, ≦60%, ≦55%, ≦50%, ≦45%, ≦40%, ≦35%, ≦30%, ≦25%, ≦20%, ≦15%, ≦10%, ≦5%, or ≦1%, of the number / proportion of such cells observed in the absence of the ADAMTS14 inhibitor or in the presence of the same amount of a control agent known to have no such inhibitory activity.

[0095] In some embodiments, an ADAMTS14 inhibitor according to the present disclosure inhibits a myofibroblast-mediated process in a given assay by less than 1-fold, e.g., one of <0.99-fold, <0.95-fold, <0.9-fold, <0.85-fold, <0.8-fold, <0.75-fold, <0.7-fold, <0.65-fold, <0.6-fold, <0.55-fold, <0.5-fold, <0.45-fold, <0.4-fold, <0.35-fold, <0.3-fold, <0.25-fold, <0.2-fold, <0.15-fold, <0.1-fold, <0.05-fold, or <0.01-fold, of the level observed in the absence of the ADAMTS14 inhibitor or in the presence of the same amount of a control agent known to lack such inhibitory activity. In some embodiments, an ADAMTS14 inhibitor according to the present disclosure inhibits a myofibroblast-mediated process in a given assay to less than 100%, e.g., one of: ≦99%, ≦95%, ≦90%, ≦85%, ≦80%, ≦75%, ≦70%, ≦65%, ≦60%, ≦55%, ≦50%, ≦45%, ≦40%, ≦35%, ≦30%, ≦25%, ≦20%, ≦15%, ≦10%, ≦5%, or ≦1%, of the level observed in the absence of the ADAMTS14 inhibitor or in the presence of an equal amount of a control agent known to have no such inhibitory activity.

[0096] In some embodiments, an ADAMTS14 inhibitor according to the present disclosure reduces collagen (e.g., type I collagen) production / deposition or extracellular matrix production / deposition or fibrosis in a given assay by less than 1-fold, for example, by one of ≦0.99-fold, ≦0.95-fold, ≦0.9-fold, ≦0.85-fold, ≦0.8-fold, ≦0.75-fold, ≦0.7-fold, ≦0.65-fold, ≦0.6-fold, ≦0.55-fold, ≦0.5-fold, ≦0.45-fold, ≦0.4-fold, ≦0.35-fold, ≦0.3-fold, ≦0.25-fold, ≦0.2-fold, ≦0.15-fold, ≦0.1-fold, ≦0.05-fold, or ≦0.01-fold, the level observed in the absence of the ADAMTS14 inhibitor or in the presence of the same amount of a control agent known to lack such inhibitory activity. In some embodiments, an ADAMTS14 inhibitor according to the present disclosure reduces collagen (e.g., type I collagen) production / deposition or extracellular matrix production / deposition or fibrosis in a given assay to less than 100%, e.g., one of ≦99%, ≦95%, ≦90%, ≦85%, ≦80%, ≦75%, ≦70%, ≦65%, ≦60%, ≦55%, ≦50%, ≦45%, ≦40%, ≦35%, ≦30%, ≦25%, ≦20%, ≦15%, ≦10%, ≦5%, or ≦1%, of the level observed in the absence of the ADAMTS14 inhibitor or in the presence of the same amount of a control agent known to have no such inhibitory activity.

[0097] Therapy and Prevention The present invention provides methods and articles of the disclosure for the treatment and / or prevention of diseases and conditions in which myofibroblasts are pathologically implicated. Treatment is achieved through inhibition of ADAMTS14, for example in myofibroblasts or myofibroblast precursor cells. Inhibition of ADAMTS14 inhibits the generation of myofibroblasts from myofibroblast precursor cells and / or the activity of myofibroblasts. Thus, the present disclosure provides an ADAMTS14 inhibitor for use in treating or preventing a disease / condition in which myofibroblasts are pathologically implicated. Also provided is the use of an ADAMTS14 inhibitor in the manufacture of a medicament for use in treating or preventing a disease / condition in which myofibroblasts are pathologically implicated. Also provided is a method of treating or preventing a disease / condition in which myofibroblasts are pathologically implicated, comprising administering a therapeutically or prophylactically effective amount of an ADAMTS14 inhibitor to a subject. It will be appreciated that the methods and articles of the disclosure may be used to treat / prevent any disease / condition that would derive therapeutic or prophylactic benefit from a reduction in myofibroblast number and / or activity. For example, the disease / condition may be one in which an increase in myofibroblast number and / or activity contributes to the pathology of the disease / condition. The disclosure also provides for the treatment / prevention of diseases / conditions caused or exacerbated by myofibroblasts.

[0098] The disease / condition may be characterized by an increase in myofibroblast number and / or activity (e.g., compared to the number / activity in the absence of the disease / condition). The disease / condition may be one in which an increase in myofibroblast number and / or activity is positively associated with the onset, development, or progression of the disease / condition. The disease / condition may be one in which an increase in myofibroblast number and / or activity is positively associated with the severity of one or more symptoms of the disease / condition. The disease / condition may be one in which an increase in myofibroblast number and / or activity is a risk factor for the onset, development, or progression of the disease / condition. The increase in myofibroblast number and / or activity according to the preceding paragraph may be in tissues and / or organs in which one or more symptoms of a disease / condition are manifested, hi some embodiments, the increase in myofibroblast number and / or activity may be in tissues and / or organs of the respiratory system, e.g., the lungs. Therapeutic or prophylactic intervention according to the present disclosure may achieve a reduction in myofibroblast number and / or activity (i.e., in the treated subject). In some embodiments, therapeutic / prophylactic intervention may achieve a reduction in myofibroblast number and / or activity in tissues and / or organs where one or more symptoms of a disease / pathology are manifested. In some embodiments, therapeutic / prophylactic intervention may achieve a reduction in myofibroblast number and / or activity in tissues and / or organs of the respiratory system, e.g., the lungs.

[0099] In some embodiments, therapeutic or prophylactic intervention according to the present disclosure is for the treatment / prevention of fibrosis. In some embodiments, the disease / condition in which myofibroblasts are pathologically involved is fibrosis or a disease / condition characterized by fibrosis. Fibrosis can be induced by pathological conditions, such as pathological conditions, infections, or disease states, that lead to the production of profibrotic factors (such as TGFβ1). Fibrosis can be caused by physical injury / stimuli, chemical injury / stimuli, or environmental injury / stimuli. Physical injury / stimuli can occur during surgery, e.g., iatrogenic causes. Chemical injury / stimuli can include drug-induced fibrosis, e.g., after chronic administration of drugs such as bleomycin, cyclophosphamide, amiodarone, procainamide, penicillamine, gold, and nitrofurantoin (Daba et al., Saudi Med J. (2004) 25(6): 700-706). Environmental injury / stimuli can include exposure to asbestos fibers or silica.

[0100] Fibrosis can be in any tissue / organ of the body. In some embodiments, fibrosis is in the lung (e.g., bronchioles, alveoli), airways (e.g., nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi), heart, kidney, liver, skeletal muscle, blood vessels, eyes, skin, pancreas, intestine, small intestine, large intestine, colon, joints, brain, or bone marrow. Fibrosis can also occur in multiple tissues / organs at once. In some embodiments, the fibrosis may be in an organ or tissue of the respiratory system, such as the lungs (e.g., bronchioles, alveoli) or airways (e.g., nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi). In some embodiments, the fibrosis may be in an organ or tissue of the cardiovascular system, such as the heart or blood vessels. In some embodiments, the fibrosis may be in an organ or tissue of the gastrointestinal system, such as the liver, intestine, small intestine, large intestine, colon, or pancreas. In some embodiments, the fibrosis may be in the eye. In some embodiments, the fibrosis may be in the skin. In some embodiments, the fibrosis may be in an organ or tissue of the nervous system, such as the brain. In some embodiments, the fibrosis may be in the bone marrow. In some embodiments, the fibrosis may be in the joints. In some embodiments, the fibrosis may be in an organ or tissue of the genitourinary system, such as the kidneys, ovaries, or fallopian tubes. In some embodiments, the fibrosis may be in an organ or tissue of the musculoskeletal system, such as muscle tissue. In some embodiments, the fibrosis may be in an organ or tissue of one or more organ systems. The present disclosure contemplates therapeutic and prophylactic interventions for diseases and conditions characterized by fibrosis. As used herein, a disease / condition "characterized by fibrosis" is a disease in which fibrosis is a symptom of the disease / condition.

[0101] Diseases and conditions characterized by fibrosis include: Diseases / conditions affecting the respiratory system, such as pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, interstitial tumors in lung diseases, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma; Diseases / conditions affecting the liver, such as chronic liver disease, liver fibrosis, cirrhosis, non-alcoholic fatty liver disease (NAFLD), steatohepatitis, non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), alcoholic fatty liver (AFL), alcoholic hepatitis, alcoholic steatohepatitis (ASH), primary biliary cirrhosis (PBC), schistosomal liver disease, and hepatocellular carcinoma (HCC); Diseases / conditions affecting the cardiovascular system, such as hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), atrial fibrosis, atrial fibrillation, ventricular fibrosis, ventricular fibrillation, myocardial fibrosis, Brugada syndrome, myocarditis, endomyocardial fibrosis, myocardial infarction, fibrotic vascular disease, hypertension, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC), atherosclerosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, varicose veins, and cerebral infarction; Diseases / conditions affecting the kidneys, such as tubulointerstitial fibrosis, glomerular fibrosis, renal fibrosis, nephritic syndrome, Alport syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry disease, diabetic nephropathy, chronic glomerulonephritis, and nephritis associated with systemic lupus erythematosus; diseases / conditions affecting the pancreas, such as pancreatic fibrosis, cystic fibrosis, and chronic pancreatitis; diseases / conditions that affect the genitourinary system, such as endometriosis; Diseases / conditions affecting the nervous system, such as gliosis, Alzheimer's disease, and multiple sclerosis; Diseases / conditions affecting the musculoskeletal system, such as muscular dystrophies, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), and fibrous myopathies; Diseases / conditions affecting the gastrointestinal system, such as inflammatory bowel disease (IBD), Crohn's disease, microscopic colitis, and primary sclerosing cholangitis (PSC); Diseases / conditions affecting the skin, such as scleroderma, nephrogenic systemic fibrosis, Dupuytren's contracture, and dermal keloids; Diseases / conditions affecting the eyes, such as Graves' ophthalmopathy, epiretinal fibrosis, retinal fibrosis, subretinal fibrosis, subretinal fibrosis associated with macular degeneration (e.g., wet age-related macular degeneration (AMD)), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis (e.g., of the posterior capsule after cataract surgery or of the bleb after trabeculectomy for glaucoma), conjunctival fibrosis, and subconjunctival fibrosis; Diseases / conditions affecting the joints, such as arthrofibrosis, arthritis, and adhesive capsulitis; Diseases / conditions affecting multiple tissue / organ systems, including progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD); fibrotic pre-neoplastic and fibrotic neoplastic diseases, and fibrosis induced by chemical or environmental insults (e.g., cancer chemotherapy, pesticides, radiation / cancer radiotherapy), Cancers such as hepatocellular carcinoma, gastric cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, and vulvar cancer; Mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease Including, but not limited to,

[0102] It will be appreciated that many of the diseases / conditions listed above are interrelated, for example, ventricular fibrosis can occur following myocardial infarction and is associated with DCM, HCM, and myocarditis. In aspects and embodiments of the present disclosure, ADAMTS14 inhibitors are provided for use in the treatment or prevention of diseases / conditions characterized by fibrosis as described herein.Also provided is the use of an ADAMTS14 inhibitor in the manufacture of a medicament for use in treating or preventing a disease / condition characterized by fibrosis as described herein.Also provided is a method of treating or preventing a disease / condition characterized by fibrosis as described herein, comprising administering a therapeutically or prophylactically effective amount of an ADAMTS14 inhibitor to a subject.

[0103] Therapeutic or prophylactic intervention according to the present disclosure may be effective to reduce the development or progression of a disease / condition, to alleviate symptoms of a disease / condition, or to reduce the pathology of a disease / condition. The intervention may be effective to prevent the progression of a disease / condition, e.g., to prevent the worsening of a disease / condition or to slow its progression. In some embodiments, the intervention may lead to an improvement of a disease / condition, e.g., a reduction in symptoms of a disease / condition, or a reduction in some other correlate of the severity / activity of a disease / condition. In some embodiments, the intervention may prevent the progression of a disease / condition to a later stage (e.g., a more severe stage, or a chronic stage). In some embodiments, the intervention may aim to slow, stop, and / or reverse the dysfunction of tissue / organ function associated with a disease / condition.

[0104] Therapeutic or prophylactic intervention according to the present disclosure may be effective in reducing the development or progression of fibrosis, alleviating fibrosis, or reducing fibrosis. Intervention may be effective in preventing the progression of fibrosis, e.g., preventing the worsening of fibrosis or slowing its rate of progression. In some embodiments, intervention may lead to an improvement, e.g., a reduction in fibrosis. In some embodiments, intervention may prevent the progression of fibrosis to a later stage (e.g., a more severe stage). In some embodiments, intervention may aim to slow, stop, and / or reverse the impairment of tissue / organ function associated with fibrosis. In aspects and embodiments of the present disclosure, ADAMTS14 inhibitors are provided for use in the treatment or prevention of fibrosis in diseases / conditions characterized by fibrosis as described herein. Use of ADAMTS14 inhibitors in the manufacture of medicaments for use in treating or preventing fibrosis in diseases / conditions characterized by fibrosis as described herein is also provided. Methods of treating or preventing fibrosis in diseases / conditions characterized by fibrosis as described herein are also provided, comprising administering a therapeutically or prophylactically effective amount of an ADAMTS14 inhibitor to a subject. Thus, in aspects and embodiments of the present disclosure, ADAMTS14 inhibitors are provided for use in the treatment or prevention of the fibrotic component of associated diseases / conditions.

[0105] Fibrosis may directly or indirectly lead to and / or increase susceptibility to certain diseases and conditions. For example, more than 80% of hepatocellular carcinoma (HCC) cases develop in fibrotic or cirrhotic livers (Affo et al. 2016, Annu Rev Pathol.), suggesting an important role for liver fibrosis in the pre-malignant environment (PME) of the liver. Therefore, the present disclosure also provides an ADAMTS14 inhibitor for use in treating and preventing diseases associated with fibrosis and / or for which fibrosis is a risk factor. In some embodiments, the disease associated with fibrosis or for which fibrosis is a risk factor is cancer, for example liver cancer (e.g., hepatocellular carcinoma). In some embodiments, the disease / condition to be treated according to the present disclosure may be characterized by an increased level of expression (i.e., gene and / or protein expression) of ADAMTS14 and / or an increased level of ADAMTS14-mediated function (e.g., compared to the number / activity in the absence of the disease / condition). The disease / condition may be a disease / condition in which an increased level of expression of ADAMTS14 and / or an increased level of ADAMTS14-mediated function is positively associated with the onset, development, or progression of the disease / condition. The disease / condition may be a disease / condition in which an increased level of expression of ADAMTS14 and / or an increased level of ADAMTS14-mediated function is positively associated with the severity of one or more symptoms of the disease / condition. The disease / condition may be a disease / condition in which an increased level of expression of ADAMTS14 and / or an increased level of ADAMTS14 function is a risk factor for the onset, development, or progression of the disease / condition.

[0106] Increased levels of ADAMTS14 expression and / or ADAMTS14-mediated function may be in cells (e.g., fibroblasts) of tissues and / or organs in which one or more symptoms of a disease / condition are manifested. In some embodiments, increased numbers and / or activity of myofibroblasts may be in cells (e.g., fibroblasts) of tissues and / or organs of the respiratory system, such as the lungs. Therapeutic or prophylactic intervention according to the present disclosure may achieve a reduction in the level of expression of ADAMTS14 and / or the level of ADAMTS14-mediated function (i.e., in the treated subject). In some embodiments, therapeutic / prophylactic intervention may achieve a reduction in the level of expression of ADAMTS14 and / or the level of ADAMTS14-mediated function in cells (e.g., fibroblasts) of tissues and / or organs in which one or more symptoms of the disease / pathology are manifested. In some embodiments, therapeutic / prophylactic intervention may achieve a reduction in the level of expression of ADAMTS14 and / or the level of ADAMTS14-mediated function in cells (e.g., fibroblasts) of tissues and / or organs of the respiratory system, such as the lungs.

[0107] In some embodiments, therapeutic or prophylactic intervention according to the present disclosure is for the treatment / prevention of fibrosis in organs / tissues of the respiratory system, e.g., selected from the lungs, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, and bronchi. In some embodiments, the therapeutic or prophylactic intervention according to the present disclosure is for the treatment / prevention of a disease / condition affecting an organ / tissue of the respiratory system, e.g., selected from pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in pulmonary disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma. In some embodiments, the therapeutic or prophylactic intervention according to the present disclosure is for the treatment / prevention of fibrosis in such disease / condition.

[0108] In some embodiments, the present disclosure provides an ADAMTS14 inhibitor for the treatment or prevention of pulmonary fibrosis (i.e., pulmonary fibrosis). In some embodiments, the present disclosure provides an ADAMTS14 inhibitor for the treatment or prevention of interstitial lung disease (ILD). In some embodiments, the present disclosure provides an ADAMTS14 inhibitor for the treatment or prevention of idiopathic interstitial pneumonia (IIP). In some embodiments, the present disclosure provides an ADAMTS14 inhibitor for the treatment or prevention of idiopathic pulmonary fibrosis (IPF). Idiopathic pulmonary fibrosis (IPF) is reviewed, for example, in Barratt et al., J Clin Med. (2018) Aug; 7(8): 201, which is hereby incorporated by reference in its entirety.

[0109] According to various aspects of the disclosure, the therapeutic / prophylactic interventions described herein include, for example, reducing expression (e.g., gene and / or protein expression) of ADAMTS14; reducing levels of RNA encoding ADAMTS14; reducing transcription of nucleic acid encoding ADAMTS14; increasing degradation of RNA encoding ADAMTS14; reducing levels of ADAMTS14 protein; increasing post-transcriptional processing (e.g., supra) of RNA encoding ADAMTS14; decreasing ADAMTS14 expression (isolating, translation, post-translational processing); increasing the degradation of the ADAMTS14 protein; decreasing the levels of correlates of ADAMTS14 activity; decreasing the cleavage of type I aminoprocollagen; decreasing one or more genes involved in fibrosis and / or metaplastic differentiation (e.g., one or more genes selected from ADAMTS14, ACTA2, COL1A1, FN1, FOXJ1, KRT5, MUC5AC, MUC5B, SCGB1A1, CTGF, CYR61, MMP7, and SOX2; e.g., reducing the expression of YAP (e.g., in fibroblasts or epithelial cells); reducing the level of YAP (e.g., in fibroblasts); reducing the nuclear localization of YAP (e.g., in fibroblasts); increasing the degradation of YAP (e.g., in fibroblasts); increasing the level of phosphorylated YAP (e.g., YAP phosphorylated at S397; e.g., in fibroblasts); reducing the level of a protein encoded by a gene whose expression is upregulated by YAP (e.g., in fibroblasts); reducing the level of TAZ (e.g., in fibroblasts); reducing the nuclear localization of TAZ (e.g., in fibroblasts); increasing the level of phosphorylated TAZ (e.g., TAZ phosphorylated at S89; e.g., in fibroblasts); reducing the level of a protein encoded by a gene whose expression is upregulated by TAZ (e.g., in fibroblasts); reducing the level of CTGF (e.g., in fibroblasts); reducing the level of CYR61 (e.g., in fibroblasts);Methods are provided for or including one or more of: reducing TGFβ1 mediated signaling (e.g., in fibroblasts); reducing levels of phosphorylated SMAD2 (e.g., in fibroblasts); reducing nuclear localization of SMAD2 (e.g., in fibroblasts); reducing levels of αSMA (e.g., in fibroblasts); reducing production of components of the extracellular matrix (e.g., type 1 collagen I; e.g., in / by fibroblasts); reducing extracellular matrix stiffening (e.g., of extracellular matrix produced by fibroblasts or epithelial cells in culture in vitro); and / or reducing stiffening of a tissue / organ (e.g., lung, liver, skin, kidney), e.g., tissue / organ involving fibrosis;

[0110] Also provided are agents according to the present disclosure for use in such methods, and the use of agents according to the present disclosure in the manufacture of compositions (e.g., medicaments) for use in such methods. It will be understood that the method includes administering an ADAMTS14 inhibitor to the subject.

[0111] Similarly, the following may be induced: decreased expression (e.g., gene and / or protein expression) of ADAMTS14; decreased levels of RNA encoding ADAMTS14; decreased transcription of nucleic acid encoding ADAMTS14; increased degradation of RNA encoding ADAMTS14; decreased levels of ADAMTS14 protein; decreased post-transcriptional processing (e.g., splicing, translation, post-translational processing) of RNA encoding ADAMTS14; increased degradation of ADAMTS14 protein; decreased levels of correlates of ADAMTS14 activity; type I amino acid protease inhibitors (type I amino acid protease inhibitors); reduced cleavage of ribonuclease; reduced expression of one or more genes involved in fibrosis and / or metaplastic differentiation (e.g., one or more genes selected from ADAMTS14, ACTA2, COL1A1, FN1, FOXJ1, KRT5, MUC5AC, MUC5B, SCGB1A1, CTGF, CYR61, MMP7, and SOX2; e.g., in fibroblasts or epithelial cells); reduced levels of YAP (e.g., in fibroblasts); reduced nuclear localization of YAP (e.g., in fibroblasts); increased degradation of YAP (e.g., in fibroblasts); increased levels of phosphorylated YAP (e.g., YAP phosphorylated at S397; e.g., in fibroblasts); decreased levels of proteins encoded by genes whose expression is upregulated by YAP (e.g., in fibroblasts); decreased levels of TAZ (e.g., in fibroblasts); decreased nuclear localization of TAZ (e.g., in fibroblasts); increased levels of phosphorylated TAZ (e.g., TAZ phosphorylated at S89; e.g., in fibroblasts); decreased levels of proteins encoded by genes whose expression is upregulated by TAZ (e.g., in fibroblasts); decreased levels of CTGF (e.g., in fibroblasts); decreased levels of CYR61 (e.g., in fibroblasts); decreased TGFβ1-mediated signaling (e.g., in fibroblasts); decreased levels of phosphorylated SMAD2 (e.g., in fibroblasts); decreased localization of SMAD2 to the nucleus (e.g., in fibroblasts); decreased levels of αSMA (e.g., in fibroblasts); decreased production of components of the extracellular matrix (e.g., type 1 collagen I; e.g., in / by fibroblasts);One or more of the following may be observed in a subject (or a tissue / organ thereof) following therapeutic or prophylactic intervention according to the present disclosure (e.g., compared to pre-intervention levels): reduced extracellular matrix stiffening (e.g., of extracellular matrix produced by fibroblasts or epithelial cells in culture in vitro); and / or reduced stiffening of a tissue / organ (e.g., lung, liver, skin, kidney), e.g., a tissue / organ containing fibrosis;

[0112] In some embodiments, a therapeutic / prophylactic intervention according to the present disclosure may be described as being "associated with" one or more of the effects described in the preceding paragraphs. Those of skill in the art can readily assess such characteristics using techniques routinely practiced in the art.

[0113] Administration of the items of the present disclosure is preferably in a "therapeutically effective" or "prophylactically effective" amount, which is sufficient to show a therapeutic or prophylactic benefit to the subject. The actual amount administered, as well as the rate and time course of administration, will depend on the nature and severity of the disease / condition and the particular item administered. Prescription of treatment, e.g., dosage, etc., is within the responsibility of general practitioners and other physicians, and typically takes into account the disease / disorder to be treated, the condition of the individual subject, the site of delivery, the method of administration, and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in Remington's 'The Science and Practice of Pharmacy' (ed. A. Adejare), 23rd Edition (2020), Academic Press. Administration of the items of the present disclosure may be parenteral, systemic, intracavitary, intravenous, intraarterial, intramuscular, intrathecal, topical, intraocular, intravitreal, intraconjunctival, subretinal, suprachoroidal, subcutaneous, intradermal, intrathecal, oral, nasal, or transdermal. Administration may be by injection, infusion, inhalation, or spray.

[0114] In some aspects and embodiments, the items of the present disclosure may be administered to the lungs. In some cases, the items of the present disclosure may be administered into the blood (i.e., intravenous / intra-arterial administration), subcutaneously, or orally. In some aspects and embodiments according to the present disclosure, there may be targeted delivery of the disclosed items, i.e., the concentration of the relevant agent in the subject is increased in a given tissue / organ compared to other parts of the body. In some embodiments, the method includes administration intravenously, intraarterially, intramuscularly, or subcutaneously, or by inhalation of aerosol, and the relevant item is formulated in a targeted agent delivery system. Suitable targeted delivery systems include, for example, nanoparticles, liposomes, micelles, beads, polymers, metal particles, dendrimers, antibodies, aptamers, nanotubes, or micro-sized silica rods. Such systems may include magnetic elements that direct the agent to the desired organ or tissue. Suitable nanocarriers and delivery systems will be apparent to those skilled in the art. In some embodiments, administration may be by inhalation of a vector encoding an ADAMTS14 inhibitor according to the present disclosure, such as an adeno-associated virus (AAV) vector or a lentivirus vector encoding an ADAMTS14 inhibitor.

[0115] In some cases, the related agent is formulated for targeted delivery to specific cells (e.g., fibroblasts), tissues and / or organs (e.g., respiratory tissues / organs, e.g., lungs). In some cases, the related agent is formulated for targeted delivery to fibroblasts. In some cases, the related agent is formulated for targeted delivery to cells of respiratory tissues / organs (e.g., fibroblasts). In some cases, the related agent is formulated for targeted delivery to lung cells (e.g., fibroblasts). The particular mode and / or site of administration may be selected according to where inhibition of ADAMTS14 is required, for example cells (eg, fibroblasts) in tissues / organs of the respiratory system (eg, lungs). In some embodiments, therapeutic or prophylactic intervention according to the present disclosure may further comprise the administration of another agent for the treatment / prevention of an associated disease / condition. The administration of the items of the present disclosure can be alone or in combination with other treatments, either simultaneously or sequentially, depending on the condition to be treated. Simultaneous administration refers to administration together with another therapeutic agent, for example as a pharmaceutical composition (combined preparation) containing both agents, or administration immediately after each other, and optionally via the same route of administration (e.g., into the same tissue, artery, vein, or other blood vessel). Sequential administration refers to administration of one agent, followed by separate administration of the other agent after a given time interval. Although this is the case in some embodiments, it is not required that the two agents be administered by the same route. The time interval can be any time interval.

[0116] Multiple dosages of the items of the present disclosure may be provided. One or more, or each, dosage may be accompanied by simultaneous or sequential administration of another therapeutic agent. The multiple doses may be separated by a predetermined time interval, which may be selected to be one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 days, or 1, 2, 3, 4, 5, or 6 months. By way of example, doses may be given once every 7, 14, 21, or 28 days (± 3, 2, or 1 day). The disclosed articles may be formulated in sustained release delivery systems to release the inhibitory nucleic acid, nucleic acid, expression vector, or composition at a predetermined rate. The sustained release delivery system may maintain a constant drug / therapeutic / prophylactic concentration for a specified period of time. In some embodiments, the disclosed articles are formulated into liposomes, gels, implants, devices, or drug-polymer conjugates, such as hydrogels.

[0117] subject The subject according to various aspects of the present disclosure may be any animal or human. Therapeutic and prophylactic applications may be in humans or in animals (i.e., in the case of veterinary use). The subject to which the article of the present disclosure is to be administered may be a subject in need of such an intervention. The subject is preferably a mammal, more preferably a human. In some embodiments, the subject may be a non-human mammal. The subject may be male or female. The subject may be a patient. The subject may have been diagnosed with a disease or condition described herein, may be suspected of having such a disease / condition, or may be at risk for developing / obtaining such a disease / condition. In embodiments according to the present disclosure, the subject may be selected for treatment according to a method based on the characterization of such disease / condition for certain markers.

[0118] Sequence identity As used herein, "sequence identity" refers to the percentage of nucleotides / amino acid residues in a subject sequence that are identical to nucleotides / amino acid residues in a reference sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage of sequence identity between the sequences. Pairwise and multiple sequence alignments for the purpose of determining percent sequence identity between two or more amino acid or nucleic acid sequences can be accomplished in a variety of ways known to those skilled in the art, for example using publicly available computer software such as ClustalOmega (Soding, J. 2005, Bioinformatics 21, 951-960), T-coffee (Notredame et al. 2000, J. Mol. Biol. (2000) 302, 205-217), Kalign (Lassmann and Sonnhammer 2005, BMC Bioinformatics, 6(298)), and MAFFT (Katoh and Standley 2013, Molecular Biology and Evolution, 30(4) 772-780) software. When using such software, default parameters, for example with respect to gap penalties and extension penalties, are preferably used.

[0119] array [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0120] Numbered paragraphs The following numbered paragraphs provide further descriptions of attributes and combinations of attributes that are contemplated in connection with the present invention. 1. ADAMTS14 inhibitors for use in the treatment or prevention of fibrosis. 2. Use of an ADAMTS14 inhibitor in the manufacture of a medicament for treating or preventing fibrosis. 3. A method of treating or preventing fibrosis in a subject, comprising administering to the subject a therapeutically or prophylactically effective amount of an ADAMTS14 inhibitor. 4. The ADAMTS14 inhibitor for use according to paragraph 1, the use according to paragraph 2, or the method according to paragraph 3, wherein the fibrosis is fibrosis of an organ or tissue of the respiratory system, lung, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, an organ or tissue of the cardiovascular system, heart, blood vessels, an organ or tissue of the gastrointestinal system, liver, intestine, small intestine, large intestine, colon, pancreas, skin, eye, an organ or tissue of the nervous system, brain, an organ or tissue of the genitourinary system, kidney, ovary, fallopian tube, an organ or tissue of the musculoskeletal system, muscle tissue, or bone marrow.

[0121] 5. Fibrosis includes pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, interstitial tumors in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, asthma, chronic liver disease, liver fibrosis, cirrhosis, nonalcoholic fatty liver disease (NAFLD), steatohepatitis, nonalcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), alcoholic fatty liver (AFL), alcohol hepatitis, alcoholic steatohepatitis (ASH), primary biliary cirrhosis (PBC), schistosomiasis liver disease, hepatocellular carcinoma (HCC), hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), atrial fibrosis, atrial fibrillation, ventricular fibrosis, ventricular fibrillation, myocardial fibrosis, Brugada syndrome, myocarditis, endomyocardial fibrosis, myocardial infarction, fibrotic vascular disease, hypertension, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC), atherosclerosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, varicose veins, cerebral infarction, tubulointerstitial fibrosis, glomerular fibrosis, renal fibrosis, nephritic syndrome, Alport syndrome, HIV Associated nephropathy, polycystic kidney disease, Fabry disease, diabetic nephropathy, chronic glomerulonephritis, nephritis associated with systemic lupus erythematosus, pancreatic fibrosis, chronic pancreatitis, endometriosis, gliosis, Alzheimer's disease, multiple sclerosis, muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), fibrous myopathy, inflammatory bowel disease (IBD), Crohn's disease, microscopic colitis, primary sclerosing cholangitis (PSC), scleroderma, nephrogenic systemic fibrosis, Dupuytren's contracture, dermal keloids, Graves' ophthalmopathy, epiretinal fibrosis, retinal fibrosis, subretinal fibrosis, jaundice Subretinal fibrosis associated with macular degeneration, wet age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis, postoperative fibrosis of the posterior capsule after cataract surgery, postoperative fibrosis of the bleb after trabeculectomy for glaucoma, conjunctival fibrosis, subconjunctival fibrosis, arthrofibrosis, arthritis, adhesive capsulitis, progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD), fibrotic preneoplastic diseases, fibrotic neoplastic diseases, fibrosis induced by chemical or environmental insult, fibrosis induced by cancer chemotherapy, fibrosis induced by pesticides,The ADAMTS14 inhibitor for use according to paragraph 1 or 4, the use according to paragraph 2 or 4, or the method according to paragraph 3 or 4, wherein the fibrosis is a disease or condition selected from radiation-induced fibrosis, cancer radiotherapy-induced fibrosis, cancer, hepatocellular carcinoma, gastric cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, vulvar cancer, mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease.

[0122] 6. The ADAMTS14 inhibitor for use according to any one of paragraphs 1, 4 or 5, the use according to any one of paragraphs 2, 4 or 5, or the method according to any one of paragraphs 3 to 5, wherein the fibrosis is fibrosis of an organ or tissue of the respiratory system, the lungs, the bronchioles, the alveoli, the airways, the nasal cavity, the oral cavity, the pharynx, the larynx, the trachea, or the bronchi. 7. The ADAMTS14 inhibitor for use according to any one of paragraphs 1 or 4 to 6, the use according to any one of paragraphs 2 or 4 to 6, or the method according to any one of paragraphs 3 to 6, wherein the fibrosis is a disease or condition fibrosis selected from pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, obliterative bronchiolitis, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma. 8. ADAMTS14 inhibitors for use in the treatment or prevention of a disease or condition characterised by fibrosis.

[0123] 9. Use of an ADAMTS14 inhibitor in the manufacture of a medicament for treating or preventing a disease or condition characterized by fibrosis. 10. A method of treating or preventing a disease or condition characterized by fibrosis in a subject, comprising administering to the subject a therapeutically or prophylactically effective amount of an ADAMTS14 inhibitor. 11. The ADAMTS14 inhibitor for use according to paragraph 8, the use according to paragraph 9, or the method according to paragraph 10, wherein the disease or condition characterized by fibrosis comprises fibrosis of an organ or tissue of the respiratory system, lung, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, an organ or tissue of the cardiovascular system, heart, blood vessels, an organ or tissue of the gastrointestinal system, liver, intestine, small intestine, large intestine, colon, pancreas, skin, eye, an organ or tissue of the nervous system, brain, an organ or tissue of the genitourinary system, kidney, ovary, fallopian tube, an organ or tissue of the musculoskeletal system, muscle tissue, or bone marrow.

[0124] 12. Diseases or conditions characterized by fibrosis include pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, interstitial tumors in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, asthma, chronic liver disease, liver fibrosis, cirrhosis, nonalcoholic fatty liver disease (NAFLD), steatohepatitis, nonalcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), and / or pulmonary fibrosis. D), alcoholic fatty liver (AFL), alcoholic hepatitis, alcoholic steatohepatitis (ASH), primary biliary cirrhosis (PBC), schistosomiasis liver disease, hepatocellular carcinoma (HCC), hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), atrial fibrosis, atrial fibrillation, ventricular fibrosis, ventricular fibrillation, myocardial fibrosis, Brugada syndrome, myocarditis, endomyocardial fibrosis, myocardial infarction, fibrotic vascular disease, hypertension, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC), atherosclerosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, varicose veins, cerebral infarction, tubulointerstitial fibrosis, glomerular Fibrosis, Renal fibrosis, Nephritic syndrome, Alport syndrome, HIV-associated nephropathy, Polycystic kidney disease, Fabry disease, Diabetic nephropathy, Chronic glomerulonephritis, Nephritis associated with systemic lupus erythematosus, Pancreatic fibrosis, Chronic pancreatitis, Endometriosis, Gliosis, Alzheimer's disease, Multiple sclerosis, Muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), Fibrous myopathy, Inflammatory bowel disease (IBD), Crohn's disease, Microscopic colitis, Primary sclerosing cholangitis (PSC), Scleroderma, Nephrogenic systemic fibrosis, Dupuytren's contracture, Dermal keryo retinal fibrosis, Graves' ophthalmopathy, epiretinal fibrosis, retinal fibrosis, subretinal fibrosis, subretinal fibrosis associated with macular degeneration, wet age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis, postoperative fibrosis of the posterior capsule after cataract surgery, postoperative fibrosis of the bleb after trabeculectomy for glaucoma, conjunctival fibrosis, subconjunctival fibrosis, arthrofibrosis, arthritis, adhesive capsulitis, progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD), fibrotic preneoplastic diseases, fibrotic neoplastic diseases, fibrosis induced by chemical insult or fibrosis induced by environmental insult,The ADAMTS14 inhibitor for use according to paragraph 8 or 11, the use according to paragraph 9 or 11, or the method according to paragraph 10 or 11, selected from cancer chemotherapy-induced fibrosis, pesticide-induced fibrosis, radiation-induced fibrosis, cancer radiotherapy-induced fibrosis, cancer, hepatocellular carcinoma, gastric cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, vulvar cancer, mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease.

[0125] 13. The ADAMTS14 inhibitor for use according to any one of paragraphs 8, 11 or 12, the use according to any one of paragraphs 9, 11 or 12, or the method according to any one of paragraphs 10 to 12, wherein the disease or condition characterized by fibrosis comprises fibrosis of an organ or tissue of the respiratory system, the lungs, the bronchioles, the alveoli, the airways, the nasal cavity, the oral cavity, the pharynx, the larynx, the trachea, or the bronchi. 14. The ADAMTS14 inhibitor for use according to any one of paragraphs 8 or 11 to 13, the use according to any one of paragraphs 9 or 11 to 13, or the method according to any one of paragraphs 10 to 13, wherein the disease or condition characterized by fibrosis is selected from pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, obliterative bronchiolitis, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in lung diseases, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma. 15. A method for inhibiting TGFβ1-mediated signaling in a cell, comprising contacting the cell with an ADAMTS14 inhibitor. 16. A method for inhibiting YAP-mediated signaling in a cell, comprising contacting the cell with an ADAMTS14 inhibitor.

[0126] 17. A method for inhibiting the production of pro-fibrotic fibroblasts, comprising contacting pro-fibrotic fibroblast precursor cells with an ADAMTS14 inhibitor. 18. A method of inhibiting a process mediated by profibrotic fibroblasts, comprising the step of contacting a profibrotic fibroblast or a profibrotic fibroblast precursor cell with an ADAMTS14 inhibitor. 19. An ADAMTS14 inhibitor for the use according to any one of paragraphs 1, 4 to 7, 8 or 11 to 14, the use according to any one of paragraphs 2, 4 to 7, 9 or 11 to 14, or the method according to any one of paragraphs 3 to 7 or 10 to 17, wherein the ADAMTS14 inhibitor reduces gene and / or protein expression of ADAMTS14. 20. An ADAMTS14 inhibitor for the use according to any one of paragraphs 1, 4 to 7, 8, 11 to 14, or 19, the use according to any one of paragraphs 2, 4 to 7, 9, 11 to 14, or 19, or the method according to any one of paragraphs 3 to 7 or 10 to 19, wherein the ADAMTS14 inhibitor is an inhibitory nucleic acid selected from siRNA, dsiRNA, miRNA, shRNA, pri-miRNA, pre-miRNA, saRNA, snoRNA, and an antisense oligonucleotide.

[0127] This disclosure includes combinations of the described aspects and preferred features except where such combinations are clearly impermissible or explicitly avoided. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Aspects and embodiments of the present disclosure will now be illustrated by way of example with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference. Throughout this specification, including the claims which follow, unless the context otherwise requires, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integers or steps or group of integers or steps. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from "about" one particular value and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment.

[0128] When a nucleic acid sequence is disclosed or referenced herein, the reverse complement thereof is also expressly contemplated. The methods described herein may preferably be carried out in vitro. The term "in vitro" is intended to include procedures that are not carried out with, in, or on a multicellular organism (e.g., procedures that are not carried out with, in, or on a human or animal body) and includes method procedures carried out with cells in culture. The term "in vivo" is intended to include procedures that are not carried out with, in, or on an intact multicellular organism (e.g., procedures that are not carried out with, in, or on a human or animal body). Embodiments and experiments illustrating the principles of the present disclosure will now be discussed with reference to the accompanying figures. [Brief description of the drawings]

[0129] [Figure 1A] 1 shows that siRNA-mediated depletion of ADAMTS14 in NHLFs significantly reduces YAP nuclear localization and also significantly reduces the expression of profibrotic genes induced by TGFβ1. Fluorescence microscopy images of cells with nuclear-localized YAP staining in NHLFs treated with control, non-targeting siRNA (siNT2), or siADAMTS14 in the presence or absence of stimulation with TGFβ1. [Figure 1B]1 is a bar graph showing that siRNA-mediated depletion of ADAMTS14 in NHLFs significantly reduced YAP nuclear localization and also significantly reduced the expression of profibrotic genes induced by TGFβ1. Quantification of the percentage of cells with nuclear-localized YAP staining in NHLFs treated with control, non-targeting siRNA (siNT2), or siADAMTS14 in the presence or absence of stimulation with TGFβ1. [Figure 1C] 1 is a bar graph showing that siRNA-mediated depletion of ADAMTS14 in NHLFs significantly reduced the nuclear localization of YAP and also significantly reduced the expression of profibrotic genes induced by TGFβ1. qPCR analysis of ADAMTS14 expression in NHLFs treated with control, non-targeting siRNA (siNT2), or siADAMTS14 in the presence or absence of stimulation with TGFβ1. [Figure 1D] 1 is a bar graph showing that siRNA-mediated depletion of ADAMTS14 in NHLFs significantly reduces the nuclear localization of YAP and also significantly reduces the expression of profibrotic genes induced by TGFβ1. Expression of profibrotic genes CTGF, CYR61, COL1A1 by NHLFs treated with control, non-targeting siRNA (siNT2), or siADAMTS14 in the presence or absence of stimulation with TGFβ1 as determined by qPCR. [Figure 2A] Figure 1 shows that treatment of NHLFs with siADAMTS14 in vitro significantly reduced YAP / TAZ protein levels and increased phosphorylation at YAPS397 and TAZS89 sites. Western blot of total YAP / TAZ protein levels after treatment of NHLFs with control siRNA (siNT2), siADAMTS14, or siADAMTS2 in the presence or absence of stimulation with TGFβ1. [Figure 2B]1 is a bar graph showing that treatment of NHLFs with siADAMTS14 in vitro significantly reduced YAP / TAZ protein levels and increased phosphorylation at YAPS397 and TAZS89 sites. Quantification of total YAP / TAZ protein levels following treatment of NHLFs with control siRNA (siNT2), siADAMTS14, or siADAMTS2 in the presence or absence of stimulation with TGFβ1. [Figure 2C] 1 is a bar graph showing that treatment of NHLFs with siADAMTS14 in vitro significantly reduced YAP / TAZ protein levels and increased phosphorylation at YAPS397 and TAZS89 sites. Quantification of phosphorylated YAP / TAZ protein levels (as a percentage of total YAP / TAZ) following treatment of NHLFs with control siRNA (siNT2), siADAMTS14, or siADAMTS2 in the presence or absence of stimulation with TGFβ1. [Figure 3A] Figure 1 shows that treatment of NHLFs with siADAMTS14 in vitro significantly reduced p-SMAD2 protein levels and inhibited TGFβ1-induced localization of SMAD2 to the nucleus. Western blot showing p-SMAD2 and SMAD2 protein expression after treatment of NHLFs with control siRNA (siNT2), siADAMTS14, or siADAMTS2 in the presence or absence of stimulation with TGFβ1. [Figure 3B] 1 is a bar graph showing that treatment of NHLFs with siADAMTS14 in vitro significantly reduced p-SMAD2 protein levels and inhibited TGFβ1-induced localization of SMAD2 to the nucleus. Quantification of p-SMAD2 (as a percentage of total SMAD2) following treatment of NHLFs with control siRNA (siNT2), siADAMTS14, or siADAMTS2 in the presence or absence of stimulation with TGFβ1. [Figure 3C]Fluorescence microscopy images of nuclear-localized SMAD2 following treatment of NHLFs with control siRNA (siNT2), siADAMTS14, or siADAMTS2 in the presence or absence of stimulation with TGFβ1. [Figure 3D] 1 is a bar graph showing that treatment of NHLFs with siADAMTS14 in vitro significantly reduced p-SMAD2 protein levels and inhibited TGFβ1-induced localization of SMAD2 to the nucleus. Quantification of nuclear-localized SMAD2 following treatment of NHLFs with control siRNA (siNT2), siADAMTS14, or siADAMTS2 in the presence or absence of stimulation with TGFβ1. [Figure 4A] 1 is an image showing that ADAMTS14 mRNA expression is significantly higher in lung biopsy samples from patients with IPF compared to lung biopsy samples from normal lungs. RNAscope® staining for ADAMTS14 mRNA. [Figure 4B] 1 is a bar graph showing that ADAMTS14 mRNA expression was significantly higher in lung biopsy samples from patients with IPF compared to lung biopsy samples from normal lungs. Quantification of the number of ADAMTS14 mRNA signals per field. [Figure 5A] 1 is a bar graph showing that treatment of human lung fibroblasts with siADAMTS14 in vitro significantly reduces TGFβ1-stimulated α-SMA fiber formation compared to TGFβ1-stimulated, non-siRNA-treated (TGFβ1) and non-stimulated, non-siRNA-treated control (CTR) cells. Data are presented ± SD from n=3 donors. [Figure 5B]1 is a bar graph showing that treatment of human lung fibroblasts with siADAMTS14 in vitro significantly reduces TGFβ1-stimulated collagen I deposition compared to TGFβ1-stimulated, non-siRNA-treated (TGFβ1) and non-stimulated, non-siRNA-treated control (CTR) cells. Data are presented ± SD from n=3 donors. [Figure 6A] Bar graph showing that treatment of lung fibroblasts with siADAMTS14 in co-culture with SAEC in vitro significantly reduces the expression of profibrotic genes. Expression of profibrotic genes ADAMTS14, ACTA2, COL1A1, and FN1 by NHLFs treated with control, non-targeting siRNA (siNT2), or siADAMTS14 in the presence or absence of stimulation with IPF-RCs as determined by qPCR. N=4 NHLF donors, *p<0.05, **p<0.01, ***p<0.001, Kruskal-Wallis / Dunn. [Figure 6B] 1 is a bar graph showing that treatment of lung fibroblasts with siADAMTS14 in co-culture with SAEC in vitro significantly reduces the expression of profibrotic genes. Expression of metaplastic differentiation / profibrotic genes FOXJ1, KRT5, MUC5AC, MUC5B, SCGB1A1, CTGF, MMP7, and SOX2 by SAEC treated with control, non-targeting siRNA (siNT2), or siADAMTS14 in the presence or absence of stimulation with IPF-RC as determined by qPCR. N=4 SAEC donors, *p<0.05, **p<0.01, ***p<0.001, Kruskal-Wallis / Dunn. [Figure 7] (A) Knockout of Adamts14 reduces lung fibrosis in mice in response to TGFβ challenge. Wild-type C57Bl / 6 mice (WT) or Adamts14− / − mice (KO) were administered AAV-TGFβ and lung collagen content was assessed 21 days later by analysis of total hydroxyproline content. *p<0.05. EXAMPLES

[0130] In this example, we demonstrate that siRNA-mediated knockdown of ADAMTS14 expression in lung fibroblasts reduces the level of YAP, inhibits YAP nuclear localization, and promotes YAP phosphorylation and degradation.Knockdown of ADAMTS14 expression in lung fibroblasts also reduces the level of TAZ, inhibits its nuclear localization and thereby its activity.It is also shown that siRNA-mediated knockdown of ADAMTS14 expression in lung fibroblasts inhibits TGFβ1-mediated signaling by reducing SMAD2 phosphorylation and SMAD2 nuclear localization.ADAMTS14 expression is found to be elevated in IPF lung tissue compared to healthy control lung tissue. It is demonstrated that inhibition of ADAMTS14 expression in lung fibroblasts inhibits the expression of profibrotic and metaplastic differentiation genes in response to stimulation with proinflammatory cytokines, as well as inhibits the formation of α-SMA fibers in and collagen I deposition by human lung fibroblasts in response to stimulation with IPF-RC.

[0131] Example 1 Materials and Methods Primary human cell lines Primary human lung fibroblasts (HLFs) from IPF or healthy control donors were harvested from de-identified discarded excess tissue from clinically indicated surgical lung resections or lung transplant explants through the MGH Fibrosis Translational Research Program. Cells were routinely grown at 37°C in a humidified incubator with 5% CO2 in DMEM (Lonza) supplemented with 10% FBS (Lonza), 2 mM L-glutamine (Lonza), 100 U / ml penicillin, and 100 μg / ml streptomycin (Lonza). Experiments were performed using fibroblasts from healthy control donor 699 unless otherwise stated. For Scar in a Jar (SiaJ) and co-culture assays, fibroblasts from Lonza (NHLF CC-2512 and DHLF-IPF CC-7231) were used. Fibroblasts were cultured in FGM-2 Fibroblast Growth Medium-2 kit (Lonza CC-3132) at 37°C in a humidified incubator with 5% CO2.

[0132] Primary siRNA screening siRNA screening and subsequent high-throughput validation experiments were performed at the ICCB-Longwood Screening Facility at Harvard Medical School. A total of 7,638 siRNA pools (Dharmacon siGENOME siRNAs) from the druggable genomic libraries Human6 and Human7 were screened in triplicate. Non-targeting siRNA controls and transfection controls were included in each plate. Primary human lung fibroblasts (HLF) from healthy donors were reverse transfected at passage 4 with 20 μM siRNA and Lipofectamine 2000 (Thermo Fisher Scientific, 11668019). Briefly, 0.05 μL Lipofectamine was diluted in 10 μL Opti-MEM (Thermo Fisher Scientific, 31985070), allowed to complex for 5 min at room temperature, and added to each well of a 384-well clear-bottom black microplate (Corning, 3764). Then, 1 μL siRNA (1 mM) was added to each well and allowed to complex for 20-30 min at room temperature. Finally, 300 cells were added to each well in 40 μL DMEM (Lonza) supplemented with 12.5% ​​FBS (Lonza) and 2 mM L-glutamine (Lonza). The plates were placed in a humidified incubator with 5% CO2 at 37°C for 72 hours and then processed for YAP imaging and localization analysis

[20] .

[0133] Secondary siRNA screening For further validation of hits, four individual siRNA duplexes comprising each siRNA pool were tested in the same assay (total of 1,512 siRNA duplexes). Targets with at least two independent active siRNA duplexes were selected for validation assays. The same four individual siRNA duplexes were retested for their ability to displace YAP and to reduce YAP target gene expression (CTGF and CYR61). Cell lysates were prepared directly on 384-well plates and gene expression analysis was performed using the Cells-to-CT 1-Step TaqMan kit (Thermo Fisher Scientific, A25602), Taqman assay (Thermo Fisher Scientific), and QuantStudio 7 Flex real-time PCR system (Thermo Fisher Scientific). 2 ΔΔCT Gene expression was calculated using the Sigma-Aldrich method and β2-microglobulin (B2M) as a housekeeping control. Results were filtered based on ability to reduce CTGF and CYR61 expression, ability to reduce nuclear / cytoplasmic YAP ratio, gene expression in HLFs (based on public and internal datasets), target novelty, druggability, and safety. The validation assay was repeated with inclusion of three additional siRNA duplexes (Ambion) and analysis of target gene expression knockdown by RT-qPCR. Two siRNA duplexes for each of the remaining hits were selected for testing in the TGFβ1 assay. Briefly, HLFs were reverse transfected as before and after 2 days were treated with or without 5ng / mL TGFβ1 in serum-free medium for 2 days. Plates were prepared for imaging (YAP and actin) and RT-qPCR analysis of profibrotic gene expression (CTGF, CYR61, COL1A1, ACTA2).

[0134] Immunofluorescence staining Cells were fixed and stained as previously described

[20] . Briefly, cells were fixed with 4% paraformaldehyde (PFA) (Electron Microscopy Sciences, 15710S) for 10-15 min and blocked with blocking solution: 10% goat serum, 0.1% Triton-X in PBS for 1 h at room temperature. Primary antibodies were incubated overnight at 4°C and secondary antibodies were incubated for 1 h at room temperature in blocking solution. The following antibodies and dilutions were used: YAP (Santa Cruz Biotechnology, sc-101199), 1:500; SMAD2 (Cell Signaling Technologies, 5339S), 1:1000; rhodamine phalloidin (Invitrogen, R415), 1:200; Alexa Fluor 488 goat anti-mouse (Thermo Fisher Scientific, A11001), 1:400; Alexa Fluor 647 goat anti-rabbit (Thermo Fisher Scientific, A-21244), 1:400. Cells in 384 and 96-well plates were stained with DAPI (Sigma-Aldrich, 100 ng / mL in PBS) and imaged with an ImageXpress confocal system (Molecular Devices).

[0135] RNAscope® Staining Staining was performed according to the manufacturer's instructions for the RNAScope® Multiplex Fluorescent V2 Assay Kit. Dilutions of probes used: RNAscope® custom designed probe ADAMTS14-C3 1:750, Opal™ 690 reagent 1:500. Images were taken using a Zeiss Axio Imager microscope and the following exposure times: DAPI: 20 ms; Cy5: 300 ms.

[0136] Cell culture and siRNA transfection Unless otherwise stated, cells were cultured at 5 × 10 3 individual cells / cm 2Cells were routinely plated at 100 ng / mL with recombinant human TGFβ1 (R&D systems, 240-B) at 5 ng / mL in serum-free medium. For knockdown of gene expression, cells were reverse transfected with Lipofectamine 2000 (Thermo Fisher Scientific, 11668019) and 20 μM siRNA according to the manufacturer's instructions. The following siRNAs from Dharmacon were used: non-targeting siRNA pool #2 (D-001206-14); siGENOME human ADAMTS14 siRNA (D-005765-01, D-005765-03, 10 μM each). After 48 h, cells were washed twice and treated with or without TGFβ1 for an additional 24 h in serum-free medium and then processed for gene expression analysis.

[0137] Western blotting Protein extracts were prepared by lysing cells on ice in RIPA buffer (Cell Signaling Technology) supplemented with Halt protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific, 78440). Protein concentration in each sample was determined using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, 23225) according to the manufacturer's instructions. Proteins were separated by SDS-PAGE (Novex® Bis-Tris 4-12% gels) using the NuPAGE® Novex® system (Thermo Fisher Scientific) under reducing conditions and transferred to nitrocellulose membranes (Thermo Fisher Scientific, LC2001). Membranes were blocked with 5% non-fat milk in TBST and all antibodies were diluted in 5% non-fat milk in TBST. Primary antibodies were incubated overnight at 4°C and secondary antibodies were added at room temperature for 30 minutes. Blots were imaged using an Odyssey® CLx imaging system. The following antibodies from Cell Signaling Technology were used at 1:1000: YAP (4912); phospho-YAP (Ser397) (13619); phospho-YAP (Ser127) (4911); TAZ (4883); phospho-TAZ (Ser89) (59971); GAPDH (2118); The following secondary antibodies were used: IRDye® 800CW goat anti-rabbit IgG secondary antibody (LI-COR, 926-32211), 1:5000; IRDye® 680RD goat anti-mouse IgG secondary antibody (LI-COR, 926-68070), 1:5000. Images were analyzed with Image Studio™ software.

[0138] RNA extraction and qPCR Cell lysates were prepared directly on the plate and gene expression was analyzed using the Cells-to-CT™ 1-Step TaqMan™ kit (Thermo Fisher Scientific, A25602). Quantitative real-time PCR was performed using the CFX Connect Real-Time PCR Detection System (Bio-Rad). The following Taqman assays from Thermo Fisher Scientific were used: CTGF Hs00170014_m1 (FAM-MGB); CYR61 Hs00155479_m1 (FAM-MGB); ACTA2 Hs00426835_g1 (FAM-MGB); COL1A1 Hs00164004_m1 (FAM-MGB); ADAMTS14 Hs01548440_m1 (FAM-MGB); B2M Hs00187842_m1 (FAM-MGB). 2 -ΔΔ Gene expression was calculated using the Ct method and β2-microglobulin (B2M) as a housekeeping control.

[0139] Scar-in-a-jar assay Normal (2 donors) and IPF-derived (1 donor) lung fibroblasts were from Lonza. siADAMTS14 was from Dharmacon (Smart Pool L-00576-00-0005). Fibroblasts were trypsinized (Reagent Pack Subculture, Lonza CC-5034) and seeded at a density of 2000 cells / 55 μl FGM-2 per well in CellCarrier-384 ultra PDL coated plates (Gibco 6057508). After 5 hours, 15 μl of siADAMTS14 / RNAiMAX (ThermoFisher 13778150) mix was added per well and incubated for 24 hours. The final concentration for siRNA was 16 nM. The medium was changed to 60 μl fibroblast basal medium (Lonza CC-3131) and incubated at 37°C, 5% CO2. After a total of 24 hours of starvation, cells were "confluent" by changing the medium to 40 μl FBM supplemented with 100 μM L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate (Sigma A8960), 37.5 mg / mL 70 kDa Ficoll (Sigma F2878), and 25 mg / mL 400 kDa Ficoll (Sigma F2637). TGF-β1 (R&D Systems 240-B) at a final concentration of 5 ng / ml was added with the confluence medium. After addition of TGF-β1, cells were incubated at 5% CO2 and 37°C for 72 hours.

[0140] Cells were washed with cold PBS and fixed with methanol for 30 min on ice. Fixed cells were washed and permeablilized with PBS-T (1% TritonX 100) for 20 min at RT. Cells were then washed and blocked with 3% BSA in PBS for 30 min at RT. After another washing step, cells were incubated with primary antibodies (anti-collagen type I (1:1000 Sigma SAB4200678) and anti-aSMA (1:1000 Sigma A2547)) for 1.5 h at 37° C., followed by 3 washes with PBS. Secondary antibodies (1:1000, goat anti-mouse IgG1 Alexa Fluor 568, Invitrogen A-21124 and goat anti-mouse IgG2a Alexa Fluor 647 Sigma SAB4600355) in PBS+Hoechst33342 (1:2000 ThermoFisher 62249) were incubated in blocking buffer at 37°C for 30 min. Samples were then washed 3 times with PBS. CellMask Green (Invitrogen H32714) was added (20 μl) to the wells at 1:50000 in PBS and incubated at RT for 30 min. For high-content imaging, an Opera Phenix High-Content Screening System (Perkin Elmer) equipped with a spinning disk confocal imaging lens was used. For fluorescence imaging, 405, 488, 568, and 647 nm lasers were used. Images were stored and analyzed in a Columbus 2.7 Image Data Storage and Analysis System (Perkin Elmer).

[0141] SAEC / NHLF coculture assay 30000 small airway epithelial cells (SAECs, Lonza CC-2547S) were seeded to confluence in PneumaCult-Ex Plus medium on 6.5 mm transwells (Corning, 0.4 μm pore size, PET membrane, 3470) coated with collagen type I (Corning). The apical medium was then removed and SAECs were allowed to stratify and differentiate at the air-liquid interface in Pneumacult-ALI-S medium (Stemcell Technologies, 05099) for 28 days. The basolateral medium was changed every other day. 25000 normal human lung fibroblasts (NHLFs, Lonza CC-2512) were seeded per well (24-well plate, Corning, 3526). After reaching 50-70% confluency, cells were starved overnight and the medium was replaced by fresh DMEM before the start of the experiment. After sufficient stratification and differentiation (28 days after ALI), SAECs were combined with NHLFs and treatment was applied basolaterally for 72 hours. Where indicated, IPF-associated cytokine cocktail (IPF-RC as described in Schruf et al. FASEB J. (2020) 34(6):7825-7846; 1:100 dilution) treatment was applied to SAEC cultures in basolateral medium 14 days after ALI, the time point at which cilia were detected.

[0142] statistical analysis Data were plotted and analyzed for statistical significance using GraphPad Prism 9 software. Two groups were compared using unpaired two-tailed t-tests, and more than two groups were compared using one- or two-way ANOVA with post-hoc tests correcting for multiple comparisons as indicated in the figure legends.

[0143] Example 2 Results and Discussion 2.1 ADAMTS14 depletion significantly reduces nuclear YAP localization and inhibits TGFβ1-induced fibroblast activation We sought to determine whether siRNA-mediated knockdown of ADAMTS14 using siADAMTS14 was associated with antifibrotic effects in normal human lung fibroblasts (NHLFs). Cells were cultured in triplicate in 96-well plates and reverse transfected with 20 μM siNT2 (si non-targeting control #2) or siADAMTS14 (individual #1 and #3 of Dharmacon siGENOME, each at 10 μM). After 48 hours, cells were washed twice and fresh serum-free medium was added with or without 5 ng / ml TGFβ1. After 24 hours, cells were fixed in 4% PFA for staining or prepared for qPCR. The results show that ADAMTS14 expression was efficiently depleted (Figure 1C) and that siADAMTS14 significantly reduced the percentage of cells with nuclear YAP localization (Figures 1A, 1B) and reduced TGFβ1-induced profibrotic gene expression (Figure 1D).

[0144] 2.2 Depletion of ADAMTS14 leads to YAP / TAZ phosphorylation and degradation After confirming that ADAMTS14 depletion leads to YAP / TAZ inactivation, we wished to determine the levels and phosphorylation status of YAP / TAZ proteins. Western blots show that total YAP protein levels were dramatically decreased after siADAMTS14 (Figures 2A, 2B). Consistently, phosphorylation of YAP at serine 397, which marks YAP for degradation in the cytoplasm, was increased after siADAMTS14 (Figure 2C). TAZ protein levels were also dramatically decreased when ADAMTS14 was depleted, but not when TGFβ1 was added, along with increased phosphorylation of TAZ at serine 89 in the absence of TGFβ1 (Figures 2A, 2C). It has been shown that phosphorylation of serine 89 in TAZ leads to sequestration of TAZ in the cytoplasm and inhibition of YAP / TAZ activity (Freeman and Morrison, Semin. Cell Dev. Biol. (2012) 23: 681-687; Irvine et al., Semin. Cell Dev. Biol. (2012) 23: 812-817). These results suggest that ADAMTS14 may have a role in regulating the phosphorylation, sequestration, and degradation of YAP and TAZ.

[0145] 2.3 siADAMTS14 inhibits the TGFβ1 signaling pathway by decreasing SMAD2 phosphorylation Because ADAMTS14 depletion has a dramatic effect in reducing TGFβ1-induced expression of profibrotic genes, we next sought to see whether canonical TGFβ1 signaling was impaired. TGFβ1 binding to its receptor leads to receptor activation, which phosphorylates SMAD2 at serine residues 465 and 467. Phosphorylated SMAD2 can enter the nucleus and promote target gene expression. NHLFs were cultured at 5 × 10 per well. 4Cells were seeded in 6-well plates at 100x and reverse transfected with 20 μM siRNA or siNT2. After 48 h, cells were washed twice and fresh serum-free medium was added with or without 5 ng / ml TGFβ1. After another 24 h, cell lysates were collected directly in the wells by lysis with RIPA buffer and resolved using SDS-PAGE. Western blot results showed that the phosphorylated form of SMAD2 was significantly decreased in HLFs after ADAMTS14 knockdown (Figure 3A, 3B). Fluorescence imaging of SMAD2 confirmed that the percentage of cells with SMAD2 localized to the nucleus was significantly decreased after siADAMTS14 (Figure 3C, 3D). These data suggest that ADAMTS14 has a role in promoting TGFβ1 canonical signaling through SMAD2 phosphorylation.

[0146] Interestingly, ADAMTS2 depletion did not have the same effect as siADAMTS14 on TGFβ1 signaling, as SMAD2 remained highly enriched in the nucleus when TGFβ1 was added to the cells (Figures 3C, 3D). This result indicates that although ADAMTS2 and ADAMTS14 are co-expressed and share a similar pool of substrates, they may have distinct roles in regulating specific signaling pathways. In the Adamts2 and Adamts14 double knockout mouse model, there is a certain skin lesion phenotype that is absent in Adamts2 single knockout mice

[21] , suggesting that ADAMTS14 may have some unknown additional role.

[0147] 2.4 ADAMTS14 mRNA expression is significantly higher in IPF lung biopsies compared with normal lungs IPF lung biopsies containing fibroblast nests were prepared by embedding in paraffin, cutting with a microtome, and mounting on slides.Using a custom-designed ADAMTS14 RNAscope® probe, we were able to detect a significantly higher number of ADAMTS14 signals per field of fibroblast nests in IPF lungs compared to normal lungs (Figure 4A, 4B).This is the first evidence that ADAMTS14 is elevated in IPF fibroblast nests and can potentially be targeted by treatment.

[0148] 2.5 ADAMTS14 depletion significantly inhibits TGFβ1-induced aSMA fibrillogenesis and collagen I deposition To determine the inhibitory efficacy of siRNA knockdown of ADAMST14 on aSMA fibrogenesis and collagen I deposition, two hallmarks of fibrogenic fibroblast-to-myofibroblast transition (FMT), data were collected from two healthy and one IPF-derived lung fibroblast cell lines. siADAMTS14 significantly inhibited TGFβ-induced α-SMA fibrogenesis by 82.2% (Figure 5A). Furthermore, collagen I deposition by lung fibroblasts was reduced by 71.2% compared to cells treated with TGFβ + transfection reagent alone (Figure 5B).

[0149] 2.6 Depletion of ADAMTS14 in fibroblasts attenuates metaplastic differentiation of airway epithelia Since fibroblasts have been shown to promote epithelial remodeling upon injury, we next assessed whether ADAMTS14-depleted fibroblasts, in turn, affect small airway epithelial cells (SAECs) during fibrotic repair. To test this, we employed air-liquid interface (ALI) culture of SAECs in co-culture with either non-targeting siRNA (siNT2) or ADAMTS14 knockdown (siADAMTS14) fibroblasts. We further exposed the co-culture system to a profibrotic environment stimulated by IPF-related cocktail (IPF-RC) for 72 hours

[22] . We first observed the induction of FMT process in NHLFs upon IPF-RC treatment (Figure 6A). As expected, depletion of ADAMTS14 attenuated IPF-RC-induced FMT. These results reinforce our observation that ADAMTS14 plays a key role in fibrotic progression in fibroblasts. Furthermore, analysis of airway epithelium revealed a dramatic increase in the expression of prometaplastic / profibrotic genes upon IPF-RC treatment (Figure 6B). Importantly, ADAMTS14-depleted fibroblasts attenuated the fibrotic effect of IPF-RC on airway epithelial lineage-involved genes. These data suggest that ADAMTS14-depleted fibroblasts in conjunction with profibrotic multifactorial stimuli act as an antifibrotic niche to block aberrant epithelial remodeling upon injury.

[0150] Example 3 Adamts14 knockout mice exhibit less fibrosis in vivo compared with wild-type mice in response to profibrotic stimuli In experiments, the inventors examined the fibrotic response in wild-type mice and in mice engineered to knock out Adamts14 expression following treatment with pro-fibrotic stimuli.

[0151] Codon usage-optimized murine cDNA encoding Tgfb1 was cloned into an AAV2-inverted terminal repeat (ITR)-containing and cytomegalovirus (CMV) promoter-containing pAAV vector. The AAV6.2 cap gene (accession number EU368910.1) was used to replace the AAV2 cap sequence in pAAV-RC. AAV6.2 vectors were produced by calcium phosphate transfection of human embryonic kidney (HEK)-293 cells using a three-plasmid-based production protocol (AAV helper-free system; number 240071; Agilent Technologies, Waldbronn, Germany) and purified. Viral titers: AAV6.2-Tgfb1: 7.26e+13vg / mL. Under light anesthesia (3–4% isoflurane), adult male wild-type C57Bl / 6 mice or Adamts14− / − mice were intubated with 50 μl of 2.7 × 10 11 vg AAV6.2-TGFb1 virus suspension was administered into the trachea. After 21 days, the mice were sacrificed and their lungs were collected for fibrosis assessment.

[0152] For the assessment of pulmonary fibrosis, total lung hydroxyproline content was determined. Briefly, harvested lungs were flash frozen and stored at -80°C until analysis. Lungs were then thawed on ice, homogenized in PBS with Halt™ protease and phosphatase inhibitors (Thermo Fisher Scientific, 78440), and hydrolyzed in 6N HCl at 120°C overnight. A 25μL aliquot was then added to 1ml of 1.4% chloramine T (MilliporeSigma), 10% n-propanol, and 0.5M sodium acetate, pH 6.0, and incubated at room temperature for 20 minutes. 1ml of Erlich's solution (1M p-dimethylaminobezaldehyde (MilliporeSigma) in 70% n-propanol, 20% perchloric acid) was then added, and the samples were incubated at 65°C for 15 minutes. The absorbance was measured at 550 nm and the amount of hydroxyproline determined against a standard curve.

[0153] Data were plotted and analyzed for statistical significance using GraphPad Prism software. Two groups were compared using a two-tailed t-test. The results are shown in Figure 7. Adamts14 knockout mice exhibited statistically significantly less pulmonary fibrosis following treatment with AAV-TGFb compared to wild-type mice, as determined by detection of reduced total hydroxyproline content.

[0154] References TIFF2024537874000006.tif197170 TIFF2024537874000007.tif94170

Claims

1. A pharmaceutical for treating or preventing fibrosis, comprising an ADAMTS14 inhibitor.

2. The pharmaceutical composition of claim 1, wherein the fibrosis is fibrosis of an organ or tissue of the respiratory system, lung, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, organ or tissue of the cardiovascular system, heart, blood vessels, organ or tissue of the gastrointestinal system, liver, intestine, small intestine, large intestine, colon, pancreas, skin, eye, organ or tissue of the nervous system, brain, organ or tissue of the genitourinary system, kidney, ovary, fallopian tube, organ or tissue of the musculoskeletal system, muscle tissue, or bone marrow.

3. Fibrosis includes pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, obliterative bronchiolitis, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, interstitial tumors in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, asthma, chronic liver disease, liver fibrosis, cirrhosis, non-alcoholic fatty liver disease (NAFLD), steatohepatitis, non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), alcoholic fatty liver (AFL), alcoholic Hepatitis, alcoholic steatohepatitis (ASH), primary biliary cirrhosis (PBC), schistosomiasis-induced liver disease, hepatocellular carcinoma (HCC), hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), atrial fibrosis, atrial fibrillation, ventricular fibrosis, ventricular fibrillation, myocardial fibrosis, Brugada syndrome, myocarditis, endomyocardial fibrosis, myocardial infarction, fibrotic vascular disease, hypertension, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC), atherosclerosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, varicose veins, cerebral infarction, tubulointerstitial fibrosis, glomerular fibrosis, renal fibrosis, nephritic syndrome, Alport syndrome, HIV-related Renal nephropathy, polycystic kidney disease, Fabry's disease, diabetic nephropathy, chronic glomerulonephritis, nephritis associated with systemic lupus erythematosus, pancreatic fibrosis, chronic pancreatitis, endometriosis, gliosis, Alzheimer's disease, multiple sclerosis, muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), fibrous myopathy, inflammatory bowel disease (IBD), Crohn's disease, microscopic colitis, primary sclerosing cholangitis (PSC), scleroderma, nephrogenic systemic fibrosis, Dupuytren's contracture, dermal keloid, Graves' ophthalmopathy, epiretinal fibrosis, retinal fibrosis, subretinal fibrosis, jaundice Subretinal fibrosis associated with macular degeneration, wet age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis, postoperative fibrosis of the posterior capsule after cataract surgery, postoperative fibrosis of the bleb after trabeculectomy for glaucoma, conjunctival fibrosis, subconjunctival fibrosis, arthrofibrosis, arthritis, adhesive capsulitis, progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD), fibrotic preneoplastic diseases, fibrotic neoplastic diseases, fibrosis induced by chemical or environmental insults, fibrosis induced by cancer chemotherapy, fibrosis induced by pesticides,2. The medicament according to claim 1, wherein the fibrosis is a disease or condition selected from radiation-induced fibrosis, cancer radiotherapy-induced fibrosis, cancer, hepatocellular carcinoma, gastric cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, vulvar cancer, mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease.

4. The pharmaceutical composition of claim 1, wherein the fibrosis is fibrosis of an organ or tissue of the respiratory system, the lung, the bronchioles, the alveoli, the airways, the nasal cavity, the oral cavity, the pharynx, the larynx, the trachea, or the bronchi.

5. 2. The pharmaceutical composition of claim 1, wherein the fibrosis is fibrosis of a disease or condition selected from pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma.

6. A pharmaceutical for treating or preventing a disease or condition characterized by fibrosis, comprising an ADAMTS14 inhibitor.

7. 7. The pharmaceutical composition of claim 6, wherein the disease or condition characterized by fibrosis comprises fibrosis of an organ or tissue of the respiratory system, lung, bronchioles, alveoli, airways, nasal cavity, oral cavity, pharynx, larynx, trachea, bronchi, organ or tissue of the cardiovascular system, heart, blood vessels, organ or tissue of the gastrointestinal system, liver, intestine, small intestine, large intestine, colon, pancreas, skin, eye, organ or tissue of the nervous system, brain, organ or tissue of the genitourinary system, kidney, ovary, fallopian tube, organ or tissue of the musculoskeletal system, muscle tissue, or bone marrow.

8. The diseases or conditions characterized by fibrosis include pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, obliterative bronchiolitis, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, interstitial tumors in lung diseases, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, asthma, chronic liver disease, liver fibrosis, cirrhosis, non-alcoholic fatty liver disease (NAFLD), steatohepatitis, non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALD), alcohol-related liver disease (ALD), and the like. Alcoholic fatty liver (AFL), alcoholic hepatitis, alcoholic steatohepatitis (ASH), primary biliary cirrhosis (PBC), schistosomiasis-induced liver disease, hepatocellular carcinoma (HCC), hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), atrial fibrosis, atrial fibrillation, ventricular fibrosis, ventricular fibrillation, myocardial fibrosis, Brugada syndrome, myocarditis, endomyocardial fibrosis, myocardial infarction, fibrotic vascular disease, hypertension, hypertensive heart disease, arrhythmogenic right ventricular cardiomyopathy (ARVC), atherosclerosis, chronic pulmonary hypertension, AIDS-related pulmonary hypertension, varicose veins, cerebral infarction, tubulointerstitial fibrosis, glomerular fibrosis, renal fibrosis, nephritis syndrome, Alport syndrome, HIV-associated nephropathy, polycystic kidney disease, Fabry disease, diabetic nephropathy, chronic glomerulonephritis, nephritis associated with systemic lupus erythematosus, pancreatic fibrosis, chronic pancreatitis, endometriosis, gliosis, Alzheimer's disease, multiple sclerosis, muscular dystrophy, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), fibrous myopathy, inflammatory bowel disease (IBD), Crohn's disease, microscopic colitis, primary sclerosing cholangitis (PSC), scleroderma, nephrogenic systemic fibrosis, Dupuytren's contracture, dermal keloids, Graves' ophthalmopathy, epiretinal fibrosis retinal fibrosis, subretinal fibrosis, subretinal fibrosis associated with macular degeneration, wet age-related macular degeneration (AMD), diabetic retinopathy, glaucoma, corneal fibrosis, postoperative fibrosis, postoperative fibrosis of the posterior capsule after cataract surgery, postoperative fibrosis of the bleb after trabeculectomy for glaucoma, conjunctival fibrosis, subconjunctival fibrosis, arthrofibrosis, arthritis, adhesive capsulitis, progressive systemic sclerosis (PSS), chronic graft-versus-host disease (GVHD), fibrotic preneoplastic diseases, fibrotic neoplastic diseases, fibrosis induced by chemical or environmental insults, fibrosis induced by cancer chemotherapy,7. The medicament according to claim 6, wherein the fibrosis is selected from pesticide-induced fibrosis, radiation-induced fibrosis, cancer radiotherapy-induced fibrosis, cancer, hepatocellular carcinoma, gastric cancer, esophageal cancer, head and neck cancer, colorectal cancer, pancreatic cancer, cervical cancer, vulvar cancer, mediastinal fibrosis, retroperitoneal fibrosis, myelofibrosis, and Peyronie's disease.

9. 7. The method of claim 6, wherein the disease or condition characterized by fibrosis comprises fibrosis of an organ or tissue of the respiratory system, the lung, the bronchioles, the alveoli, the airways, the nasal cavity, the oral cavity, the pharynx, the larynx, the trachea, or the bronchi.

10. 7. The pharmaceutical composition of claim 6, wherein the disease or condition characterized by fibrosis is selected from pulmonary fibrosis, interstitial lung disease (ILD), idiopathic interstitial pneumonia (IIP), idiopathic pulmonary fibrosis (IPF), cystic fibrosis, progressive massive fibrosis, scleroderma, bronchiolitis obliterans, Hermansky-Pudlak syndrome, asbestosis, silicosis, sarcoidosis, tumor interstitium in lung disease, chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, and asthma.

11. A method for inhibiting TGFβ1-mediated signaling or YAP-mediated signaling in a cell, comprising contacting the cell with an ADAMTS14 inhibitor.

12. A method of inhibiting the production of pro-fibrotic fibroblasts, comprising contacting pro-fibrotic fibroblast precursor cells with an ADAMTS14 inhibitor.

13. A method of inhibiting a process mediated by profibrotic fibroblasts, comprising contacting profibrotic fibroblasts or profibrotic fibroblast precursor cells with an ADAMTS14 inhibitor.

14. The pharmaceutical agent according to claim 1 or claim 6, or the method according to any one of claims 11 to 13, wherein the ADAMTS14 inhibitor reduces ADAMTS14 gene and / or protein expression.

15. The pharmaceutical composition according to claim 1 or claim 6, or the method according to any one of claims 11 to 13, wherein the ADAMTS14 inhibitor is an inhibitory nucleic acid selected from siRNA, dsiRNA, miRNA, shRNA, pri-miRNA, pre-miRNA, saRNA, snoRNA, and antisense oligonucleotides.

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