Col12a1 as a therapeutic target in uterine leiomyomas

Abstract

Novel methods of treatment of uterine leiomyomas is presented. FACIT collagens, such as COL12A1, COL22A1, and COL9A2 were found to be overexpressed in uterine leiomyomas. COL12A1 was found to be a dual function structural and signaling mediator. Targeting COL12A1 offers a new target for non-surgical treatment of uterine leiomyomas.

Description

C0L12A1 AS A THERAPEUTIC TARGET IN UTERINE LEIOMYOMAS CROSS REFERENCE TO RELATED APPLICATIONSThis application is a nonprovisional of and claims priority to U S Provisional Patent Application Serial No. 63 / 763,888, filed February 26, 2025, the contents of which are hereby incorporated by reference into this disclosure.SEQUENCE LISTINGThe sequence listing entitled “COL12A1 as a Therapeutic Target in Uterine Leiomyomas” in XML format, created on February 26, 2026, and being 2000 bytes in size, is hereby incorporated by reference into this disclosure.FIELD OF INVENTIONThis invention relates to methods of treatment and / or prevention of leiomyomas Specifically, the invention provides a novel methods of treating and preventing growth of uterine leiomyomas by inhibiting expression of COL12A1BACKGROUND OF THE INVENTIONUterine leiomyomas are neoplastic proliferations found in as many as 70% of reproductive-age women. A frequent cause of abnormal uterine bleeding, pelvic pain, and infertility, the economic burden attributed to leiomyomas in the U.S. is estimated to range between $6 and $34 billion annually.Current clinical management of leiomyomas relies on surgical and hormonal interventions used to control symptoms and / or restore reproductive tract function. Hormonal antagonists and selective progesterone receptor modulators, such as ulipristal, are often used to control vaginal bleeding. However, these strategies are often less effective at alleviating pelvic pain and symptoms arising from local mass effect Due to their ability to disrupt ovulation, they also typically impair fertility Conservative organ-preserving procedures, such as myomectomy, can also be used to selectively excise or ablate tumor. However, recurrences following these procedures are common, and as many as 57% women experiencing a recurrence within 5 years. Alternative, non-surgical procedures, such as radiofrequency ablation, are also an option, but are characterized by substantial failure rates that frequentlynecessitate hysterectomy. As a result, leiomyomas remain the most common indication for hysterectomy in the United States (U.S.).Histologically, the accumulation of extracellular matrix (ECM) is a well-established hallmark of uterine leiomyomas. Abundant expression of fibrillar collagens, including Types I, III, and V, is driven by steroid hormones, the overexpression of growth factors, such as TGF-p and PDGF and imbalances in the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). This results in a disorganized extracellular matrix in leiomyomas that is histologically distinct from that found in adjacent healthy myometrium. By multiple measures, leiomyomas are stiffer than adjacent healthy myometrium. Robust data indicate that mechanical stress resulting from this stiffness plays an active role in promoting leiomyoma growth. However, the mechanisms by which the extracellular microenvironment promotes the growth of these tumors remains poorly understood. Furthermore, the role of other types of collagens with important roles in extracellular signaling, such as fibril-associated collagens with interrupted triple helices (FACITs) remain unexplored.In contrast to fibrillar collagens, FACIT collagens are characterized by structural interruptions in their helical structure. In other cells and tissues, FACIT collagens play an important role in cross-linking and organizing collagen strands in the ECM High levels of FACIT collagens, including Types IX (COL9A1), XI (COL11A1) and XII (COL12A1), are found in tissues exposed to mechanical stress, such as cartilage. There, they decorate the surface of fibrillar collagens and act as molecular bridges to cells and other components of the extracellular matrix, such as proteoglycans. Pathogenic mutations in COL12A1 have been described in a clinical subset of Ehlers-Danlos Syndrome known as myopathic EDS and clinically result in joint laxity. Functional studies have also reported that collagen XII plays an active role in wound healing and scar formation. More recently, collagen XII has been proposed as a non-invasive marker of activated fibroblasts in solid tumors, further supporting its relevance to tumorigenic processes.In view of the lack of non-surgical efficacious treatments for uterine leiomyomas, what is needed is an effective, non-surgical option for treating leiomyomas that preserves fertilitySUMMARY OF INVENTIONThe inventors have characterized patterns of FACIT collagens in human uterine leiomyomas and found that overexpression of COL12A1 and multiple other collagens (COL9A2 and COL22A1) with interrupted triple helices (FACITs) are a robust feature of uterine leiomyomas with robust Type XII collagen (COL12A1) overexpression being a consistent feature of leiomyomas. Interrogation of transcripts generated by single cell profiling indicates that COL12A1 is robustly expressed in diverse cell types in leiomyomas and mediates communication between uterine myocytes, myofibroblasts, macrophages, myeloid cells, and endothelium In primary cultures derived from human leiomyoma, functional silencing of COL12A1 reduces proliferation, promotes G1-S cell cycle arrest, and inhibits AKT1-mediated signaling. Targeting COL12A1 expression disrupts the morphological compaction and survival of three-dimensional leiomyoma organoids promoted by mechanical stress and dramatically alters the abundance and histologic features of organoid cell populations. Collectively, these findings establish COL12A1 as a novel, dual function structural and signaling mediator that integrates ECM remodeling and mechanical stress to drive leiomyoma growth and identify a new target for treatmentIn an aspect, a method of treating uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least gene or gene expression product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs) In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is premenopausal. The therapeutically effective amount of the composition acts to downregulate expression of at least one gene or gene expression product encoding at least one FACIT in the uterine leiomyoma in the patient to treat the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In a preferable aspect, the at least one FACIT is collagen XII. In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof. In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1. In certain aspects targeting COL12A1 downregulation, the therapeutic agent may comprise an RNA interference agent targeting the gene or gene expression product, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and / or combinationsthereof. The therapeutically effective amount of the composition downregulates expression of the at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to treat the uterine leiomyoma. In an aspect, a method of treating uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production / synthesis of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT) In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof The therapeutically effective amount of the composition reduces the amount of the at least one FACIT in the uterine leiomyoma in the patient to treat the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII In certain aspects, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In a further aspect, a method of inhibiting growth of uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition is sufficient to downregulate expression of at least one gene or gene expression product encoding at least one FACIT in the uterine leiomyoma in the patient to inhibit growth of the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In a preferable aspect, the at least one FACIT is collagen XII. In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1chain (COL22A1), and combinations thereof. In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1. In certain aspects targeting COL12A1 downregulation, the therapeutic agent may comprise an RNA interference agent targeting the gene or gene expression product, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and / or combinations thereof.In an aspect, a method of inhibiting growth of uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production / synthesis of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. The therapeutically effective amount of the composition reduces the amount of the at least one FACIT in the uterine leiomyoma in the patient to inhibit growth of the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII In certain aspects, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In a further aspect, a method of inhibiting tumor cell proliferation in uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition downregulates expression of at least one gene or gene expression product encoding at least one FACIT in the uterine leiomyoma in the patient to inhibit tumor cell proliferation in the uterine leiomyoma. In certainaspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In a preferable aspect, the at least one FACIT is collagen XII. In certain aspects, the at least one gene or gene expression product being downregulated comprises collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1. In certain aspects targeting COL12A1 downregulation, the therapeutic agent may comprise an RNA interference agent targeting the gene or gene expression product, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and / or combinations thereofIn an aspect, a method of inhibiting tumor cell proliferation in uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. The therapeutically effective amount ofthe composition reduces the amount of the at least one FACIT in the uterine leiomyoma in the patient to inhibit tumor cell proliferation in the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII In certain aspects, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In further aspects, a method of decreasing (reducing) accumulation of extracellular matrix (ECM) in uterine leiomyomas in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least gene encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, thecomposition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition downregulates expression of at least one gene or gene expression product encoding at least one FACIT in the uterine leiomyoma in the patient to decrease (reduce) ECM accumulation in the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In a preferable aspect, the at least one FACIT is collagen XII. In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof. In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1 In certain aspects targeting COL12A1 downregulation, the therapeutic agent may comprise an RNA interference agent targeting the gene or gene expression product, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and / or combinations thereofIn certain aspects, a method of decreasing (reducing) accumulation of extracellular matrix (ECM) in uterine leiomyomas in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production / synthesis of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). The therapeutically effective amount of the composition reduces the production / synthesis or amount of the at least one FACIT in the uterine leiomyoma in the patient to decrease accumulation of ECM in the uterine leiomyoma. In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain preferred aspects, the FACIT is COL12A1. In certain aspects, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In further aspects, a method of preventing fibrosis in uterine leiomyomas in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least gene encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition downregulates expression of at least one gene or gene expression product encoding at least one FACIT in the uterine leiomyoma in the patient to prevent fibrosis in the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In a preferable aspect, the at least one FACIT is collagen XII. In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1 In certain aspects targeting COL12A1 downregulation, the therapeutic agent may comprise an RNA interference agent targeting the gene or gene expression product, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and / or combinations thereofIn certain aspects, a method of preventing fibrosis in uterine leiomyomas in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production / synthesis of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). The therapeutically effective amount of the composition reduces the production / synthesis or amount of the at least one FACIT in the uterine leiomyoma in the patient to prevent fibrosis in the uterine leiomyoma. In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain preferred aspects, the FACIT is COL12A1. In certain aspects, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, ornilotinib. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In further aspects, a method of reducing structural integrity and / or reducing stiffness in uterine leiomyomas in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least gene encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition downregulates expression of at least one gene or gene expression product encoding at least one FACIT in the uterine leiomyoma in the patient to reduce structural integrity and / or reduce stiffness in the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof In a preferable aspect, the at least one FACIT is collagen XII. In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof. In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1 In certain aspects targeting COL12A1 downregulation, the therapeutic agent may comprise an RNA interference agent targeting the gene or gene expression product, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and / or combinations thereof.In certain aspects, a method of reducing structural integrity and / or reducing stiffness in uterine leiomyomas in a patient in need thereof comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). The therapeutically effective amount of the composition reduces the prod uction / sy nth esis or amount of the at least one FACIT in the uterine leiomyoma in the patient to reduce structural integrity and / or reduce stiffness the uterine leiomyoma. In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain(C0L12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain preferred aspects, the FACIT is COL12A1. In certain aspects, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the patient is pre-menopausal In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.BRIEF DESCRIPTION OF THE DRAWINGSFor a fuller understanding of the invention, reference should be made to the following detailed description, taken in connection with the accompanying drawings, in which:FIG. 1 A is a series of images depicting FACIT expression in healthy myometrium and uterine leiomyoma. (A) Expression of specific FACIT collagens were evaluated in matched specimens of healthy myometrium (Myo) and leiomyoma (Leio) by GeneChip. Elevated COL12A1 expression was observed in specimens of leiomyoma regardless of whether specimens were collected in either the proliferative (Pro; n=12; p= 00004) and secretory (Sec; n=9; p=0.007) phase of the menstrual cycle. Results are reported as mean expression + SD.FIG. 1 B is a series of images depicting FACIT expression in healthy myometrium and uterine leiomyoma. (B) Immunohistochemical evaluation of COL12A1 expression in formalin-fixed specimen of human leiomyoma. Bar represents 100 microns. COL12A1 expression was evaluated in cell pellets and supernatants of primary cultures derived from matched specimens of myometrium and leiomyoma (n=6 each).FIG. 1C-E is a series of graphs depicting FACIT expression in healthy myometrium and uterine leiomyoma. (C) Expression of COL12A1 transcript evaluated in primary cultures derived from specimens of healthy myometrium and leiomyoma by RT-qPCR; (D) COL12A1 expression in primary cultures evaluated by ELISA. (E) Evaluation of COL12A1 expression supernatant of primary cultures by ELISA. FIG.1F-G are images depicting FACIT expression in healthy myometrium and uterine leiomyoma. (F) Western blot evaluating COL12A1 expression in cell pellets generated from primary cultures (n=3) (G) Densitometric evaluation of Western blotimages confirms COL12A1 expression. Results (C-G) are reported as mean intensity + standard deviation. *p<0.05; **p<0.01; ***p<0.001.FIG. 2A is an image depicting biomarker defined cell populations expressing COL12A1 in healthy myometrium and leiomyoma. (A) Relative expression of COL12A1 in biomarker-defined populations of cells identified by single-cell RNA-seq performed on matched specimens of healthy myometrium shown in (A) and leiomyoma shown in (B).FIG. 2B is an image depicting biomarker defined cell populations expressing COL12A1 in healthy myometrium and leiomyoma. (B) Relative expression of COL12A1 in biomarker-defined populations of cells identified by single-cell RNA-seq performed on leiomyoma specimens.FIG. 2C is an image depicting biomarker defined cell populations expressing COL12A1 in healthy myometrium and leiomyoma. (C) Delineation of signaling dyads between these cell populations was evaluated by CellChat. These analyses reveal that communication between these populations of cell differ significantly between healthy myometrium shown in (C) and leiomyoma shown in (D). Arrowheads indicated directionality of potential paracrine signaling while thickness of stem is proportional to number of distinct signaling pathways identified.FIG. 2D is an image depicting biomarker defined cell populations expressing COL12A1 in healthy myometrium and leiomyoma. (D) Delineation of signaling dyads between these cell populations was evaluated by CellChat. These analyses reveal that communication between these populations of cell differ significantly between healthy myometrium shown in (C) and leiomyoma shown in (D). Arrowheads indicated directionality of potential paracrine signaling while thickness of stem is proportional to number of distinct signaling pathways identified.FIG.3A-C are a series of images depicting targeting COL12A1 expression in primary cultures of leiomyoma suppresses proliferation and induces G1 cell cycle arrest in leiomyoma cells. (A, B, C) Proliferation was evaluated in primary cultures derived from leiomyoma transfected with either an siRNA targeting COL12A1 expression (red) or a scrambled nontargeting (blue) in in three independent leiomyoma lines (Leio 58, 94, 102) These results clearly demonstrate that siRNA-mediated knockdown of COL12A1 significantly reduced proliferation over the course of 72 hours.FIG. 3D is a series of images depicting targeting C0L12A1 expression in primary cultures of leiomyoma suppresses proliferation and induces G1 cell cycle arrest in leiomyoma cells. (D) Targeting COL12A1 expression resulted in a reduced proportion of cells in S-phase and G-2M phases of the cell cycle also when these cell lines. FIG. 3E is a series of images depicting targeting COL12A1 expression in primary cultures of leiomyoma suppresses proliferation and induces G1 cell cycle arrest in leiomyoma cells. (E) Targeting COL12A1 expression resulted in a reduced proportion of cells in S-phase and G-2M phases of the cell cycle also when these cell lines. FIG. 3F is a series of images depicting targeting COL12A1 expression in primary cultures of leiomyoma suppresses proliferation and induces G1 cell cycle arrest in leiomyoma cells. (F) Targeting COL12A1 expression resulted in a reduced proportion of cells in S-phase and G-2M phases of the cell cycle also when these cell lines. FIG. 3G-L is a series of images depicting targeting COL12A1 expression in primary cultures of leiomyoma suppresses proliferation and induces G1 cell cycle arrest in leiomyoma cells. Successful targeting of COL12A1 expression was confirmed both by RT-qPCR (G, H, I) and ELISA (H, J, L) in each cell line evaluated. For ELISA, results are reported as ng COL12A1 / ml. *p<0.05; **p<0.01.FIG. 4A-C is a series of images depicting targeting COL12A1 progressively impacts the compaction and integrity of leiomyoma organoids in vitro. (A) Brightfield microscopy was used to evaluate spheroids generated from cell suspensions of leiomyoma cultures following transfection with either shRNA targeting COL12A1 (shCOL12A1) or a scrambled, non-targeting control (shCon) (n=3). These analyses revealed that targeting COL12A1 expression resulted in organoids with increased cross section area (B, pm2) but contained fewer cells (C) as early as 24 hours after transfer to organoid media.FIG. 4D-F is a series of images depicting targeting COL12A1 progressively impacts the compaction and integrity of leiomyoma organoids in vitro. (D) Over time, organoids generated from leiomyomas cells in which COL12A1 had been targeted progressively fail to organize and undergo compaction typical observed in control cultures. Targeting COL12A1 expression also resulted in the loss of elongated epithelo id-l I ke cells that typically line the surface of these organoids (E), as well as a dramatic accumulation of numerous lipid droplets in myofibroblast-like cells found within organoid interior (F).FIG. 5A-D is a series of images depicting C0L12A1 plays a key role in mediating the impact of mechanical stress on organoid integrity and compaction. (A) Global application of mechanical stress to primary cultures of healthy myometrium results in more compact organoids within 24 hours of plating. This is evidenced by significantly smaller cross-sectional area (B, pm2) that contain greater numbers of cells as evidenced by DAPI staining (C). (D) Application of mechanical stress to 2-dimensional cultures of healthy myometrium (Myo#82, Myo#95, Myo#106) results in significantly increased COL12A1 expression when evaluated by RT-qPCR.FIG. 5 E-G is a series of images depicting COL12A1 plays a key role in mediating the impact of mechanical stress on organoid integrity and compaction. (E) Application of mechanical stress results in increased luciferase activity in primary cultures of healthy myometrium (Myo#4, Myo #106, Myo#8) transfected with transfected with reporter vector in which luciferase activity is regulated by the consensus promoter for COL12A1. In contrast, mechanical stress had no impact on luciferase activity in cultures transected with either empty vector (Vector) lacking the coding sequence for luciferase or a second control in which luciferase transcription is regulated by minimal promoter element (MinP). (F and G) Mechanical stress had no impact on the compaction of organoids generated from primary leiomyoma cultures transfected with shCOL121 *p<0.05; “p<0.01.FIG. 6A-B is a series of images depicting biological impact of targeting COL12A1 expression on primary cultures derived from human leiomyomas. (A) Western blot was used to compare levels of expression for AKT1, phospho-AKT1 and Cyclin D in primary leiomyoma cultures (Leio#85, Leio#94 and Leio#102) transfected with either siRNA targeting COL12A1 expression (siCOL12A1) or a scrambled, non-targeting control (siCON). (B) Densitometry of Western blots confirms that successful knockdown of COL12A1 expression results in significantly lower levels of phoepho-AKT1 and cyclin D expression. In contrast, application of mechanical stress (MG) results in significantly increased expression of p-AKT1 and cyclin D1.FIG. 6C-D is a series of images depicting biological impact of targeting COL12A1 expression on primary cultures derived from human leiomyomas. (C) Western blot evaluating expression for AKT1, phospho-AKT1 and Cyclin D in primary cultures of healthy myometrium exposed to mechanical stress (MG) and matched controls. (D) Densitometry of Western blots confirms that exposure to mechanical stress results in significantly greater levels of phospho-AKT 1 and cyclin D expression when compared to sham-treated controls (n=3). After normalization to the corresponding GAPDHcontrol, results were quantified and reported as mean intensity ± SD *p<0.05 vs. control, **p<0.01 vs. controlDETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTIn the following detailed description of the preferred embodiments, reference is made to the accompanying drawings, which form a part hereof, and within which are shown by way of illustration specific embodiments by which the invention may be practiced. It is to be understood that other embodiments may be utilized, and structural changes may be made without departing from the scope of the invention.AbbreviationsCOL9A1 - Type IX collagen alpha chainCOL11 A1 - Type XI collagen alpha chainCOL12A1 - Type XII collagen alpha chainECM - extra cellular matrixFACITs - fibril-associated collagens with interrupted triple helicesMMPs - matrix metalloproteinasesTIMPs - tissue inhibitors of metalloproteinasesDefinitionsUnless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, some potential and preferred methods and materials are described herein. All publications mentioned herein are incorporated herein by reference in their entirety to disclose and describe the methods and / or materials in connection with which the publications are cited It is understood that the present disclosure supersedes any disclosure of an incorporated publication to the extent there is a contradiction. As used herein, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise.As used in this specification and the appended claims, the term “or” is generally employed in its sense including “and / or” unless the context clearly dictates otherwise. All numerical designations, such as pH, temperature, time, concentration, and molecular weight, including ranges, are approximations which are varied up ordown by increments of 1.0, 0.1, 0.01 or 0.001 as appropriate. It is to be understood, even if it is not always explicitly stated that all numerical designations are preceded by the term “about.” It is also to be understood, even if it is not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art and can be substituted for the reagents explicitly stated herein. Concentrations, amounts, solubilities, and other numerical data may be expressed or presented herein in a range format. It is to be understood that such a range format is used merely for convenience and brevity and thus should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. As an illustration, a numerical range of “about 1 to about 5” should be interpreted to include not only the explicitly recited values of about 1 to about 5 but also include the individual values and sub-ranges within the indicated range. Thus, included in this numerical range are individual values such as 2, 3, and 4 and sub-ranges such as from 1-3, from 2-4 and from 3-5, etc. This same principle applies to ranges reciting only one numerical value. Furthermore, such an interpretation should apply regardless of the range or the characteristics being describedAs used herein, the term “comprising” is intended to mean that the products, compositions, and methods include the referenced components or steps, but not excluding others. “Consisting essentially of when used to define products, compositions, and methods, shall mean excluding other components or steps of any essential significance that affect the novel characteristics of the invention as described herein. Thus, a composition consisting essentially of the recited components would not exclude trace contaminants and pharmaceutically acceptable carriers. “Consisting of” shall mean excluding more than trace elements of other components or steps.As used herein, “about” means approximately or nearly and in the context of a numerical value or range set forth means ±10% of the numerical.As used herein “patient” is used to describe a mammal, preferably a human, to whom treatment is administered, including prophylactic treatment with the compositions of the present invention. Non-limiting examples of mammals include humans, rodents, aquatic mammals, domestic animals such as dogs and cats, farm animals such as sheep, pigs, cows, and horses. “Patient” and “subject” are used interchangeably herein.“Administering” or “administration” as used herein refers to the process by which the compositions of the present invention are delivered to the patient. The compositions may be administered in various ways, including but not limited to, orally, mucosally, percutaneously, subcutaneously, and parenterally, although other enteral and parenteral routes are contemplated. Any of the compounds may also be delivered through encapsulation in vesicles such as liposomes, niosomes, micelles, etc. “Parenteral administration” as used herein refers to modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intra-articular, subcapsular, intrathecal, intraventricular, intracisternal, intranigral, subarachnoid, intraspinal, and intrasternal injection and infusion. Dosing can be by any suitable route, e.g., by injections, such as intravenous or percutaneous injections, depending in part on whether the administration is brief or chronic. Various dosing schedules including but not limited to, single or multiple administrations over various time-points, bolus administration, and pulse infusion are contemplated herein. A “therapeutic agent” as used herein refers to a substance, composition, compound, chemical, component, or extract that has measurable specified or selective physiological activity when administered to an individual in a therapeutically effective amount. In some embodiments, the therapeutic agent may be a small molecule or RNA interference agent such as a shRNA, siRNA, or anti-sense oligonucleotide, targeting the at least one gene or gene expression product selected from COL12A1, COL22A1 , COL9A2 or combinations thereof. Examples of therapeutic agents as used in the present invention include, but are not limited to, therapeutic agents which reduce expression of genes or gene expression products encoding production of FACITs such as COL9A2, COL12A1, and / or COL22A1 as well as therapeutic agents which reduce the amount of or production of at least one FACIT such as COL9A2, COL12A1, and / or COL22A1. In certain aspects, the therapeutic agent reduces expression of COL12A1. In certain aspects, the therapeutic agent reduces production of COL12A1. At least one therapeutic agent is used in the compositions of the presentinvention, however in some embodiments, multiple therapeutic agents are used. In some embodiments, the therapeutic agent targeting COL12A1 expression, production or amount described herein may be combined with another therapeutic agent that targets a different disease target. In some embodiments, one or more therapeutic agents may be encapsulated within a carrier vesicle such as a nanoparticle or micelle or other lipid carrier. In some embodiments, the therapeutic agent is used to treat leiomyomas.The terms “reduce or inhibit” as used herein refers to the ability to cause an overall decrease of 10%, 20%. 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or greater, including all intervening amounts. Reduce or inhibit can refer, for example, to the occurrence and / or frequency and / or severity of symptoms of the disease being treated, to the amount of damage or injury incurred by the patient from the disorder being treated, to the amount of leiomyoma growth, to the amount of tumor cell proliferation, to the amount of stiffness of the uterine leiomyoma, to the accumulation of ECM in uterine leiomyomas, to the production of FACITs, to the amount of FACITs, etc.A “therapeutically effective amount” as used herein is defined as concentrations or amounts of components which are sufficient to effect beneficial or desired clinical results, including, but not limited to, any one or more of treating symptoms of uterine leiomyomas, and preventing uterine leiomyomas. Compositions of the present invention can be used to effect a favorable change in the condition whether that change is an improvement, such as stopping, reversing, or a complete elimination of symptoms due to the disorder. In accordance with the present invention, a suitable single dose size is a dose that is capable of preventing or alleviating (reducing or eliminating) a symptom in a patient when administered one or more times over a suitable time period. One of skill in the art can readily determine appropriate single dose sizes for systemic administration based on the size of the animal and the route of administration. The dose may be adjusted according to response.The dosing of compounds and compositions to obtain a therapeutic or prophylactic effect is determined by the circumstances of the patient, as is known in the art. The dosing of a patient herein may be accomplished through individual or unit doses of the compounds or compositions herein or by a combined or prepackaged or preformulated dose of a compounds or compositions.The amount of the compound in the drug composition will depend on absorption, distribution, metabolism, and excretion rates of the drug as well as other factorsknown to those of skill in the art. Dosage values may also vary with the severity of the condition to be alleviated. The compounds may be administered once or may be divided and administered over intervals of time. It is to be understood that administration may be adjusted according to individual need and professional judgment of a person administrating or supervising the administration of the compounds used in the present invention.The dose of the compounds administered to a subject may vary with the particular composition, the method of administration, and the particular disorder being treated. The dose should be sufficient to affect a desirable response, such as a therapeutic or prophylactic response against a particular disorder or condition. It is contemplated that one of ordinary skill in the art can determine and administer the appropriate dosage of compounds disclosed in the current invention according to the foregoing considerations.Dosing frequency for the composition includes, but is not limited to, at least about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week, or daily. In some embodiments, the interval between each administration is less than about a week, such as less than about any of 6, 5, 4, 3, 2, or 1 day. In some embodiments, the interval between each administration is constant. For example, the administration can be carried out daily, every two days, every three days, every four days, every five days, or weekly. In some embodiments, the administration can be carried out twice daily, three times daily, or more frequently. Administration can also be continuous and adjusted to maintaining a level of the compound within any desired and specified range.The administration of the composition can be extended over an extended period of time, such as from about a week or shorter up to about a year or longer. For example, the dosing regimen can be extended over a period of any of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 months. In some embodiments, there is no break in the dosing schedule. In some embodiments, the interval between each administration is no more than about a week.The therapeutic agents used in the present invention may be administered individually, or in combination with or concurrently with one or more other therapeutic agents used against uterine leiomyomas. Additionally, therapeutic agents used in the present invention may be administered in combination with or concurrently with other therapeutics for uterine leiomyomas.“Prevention” or “preventing” or “prophylactic treatment” as used herein refers to any of: halting the effects of uterine leiomyomas, reducing the effects of uterine leiomyomas, reducing the incidence of uterine leiomyomas, reducing the development of uterine leiomyomas, delaying the onset of symptoms of uterine leiomyomas, increasing the time to onset of symptoms of uterine leiomyomas, and reducing the risk of development of uterine leiomyomas.“Treatment” or “treating” as used herein refers to any of the alleviation, amelioration, elimination, and / or stabilization of a symptom, as well as delay in progression of a symptom of a particular disease or disorder, particularly uterine leiomyomas. For example, “treatment” of a uterine leiomyoma may include any one or more of the following: amelioration and / or elimination of one or more symptoms associated with uterine leiomyomas, reduction of one or more symptoms of uterine leiomyomas, stabilization of symptoms of uterine leiomyomas, and delay in progression of one or more symptoms of uterine leiomyomas.The pharmaceutical compositions of the instant invention may comprise sufficient genetic material to produce a therapeutically effective amount of the compound of interest, i. e. , an amount sufficient to reduce or ameliorate symptoms of the uterine leiomyoma, or an amount sufficient to confer the desired benefit. The pharmaceutical compositions of the subject invention can be formulated according to known methods for preparing pharmaceutically useful compositions. When the therapeutic agents of the invention are prepared for administration, they are preferably combined with a pharmaceutically acceptable carrier, diluent, or excipient to form a pharmaceutical formulation, or unit dosage form. The total active ingredients in such formulations include from 0.1 to 99.9% by weight of the formulation. Furthermore, as used herein, the phrase “pharmaceutically acceptable carrier” means any of the standard pharmaceutically acceptable carriers. The pharmaceutically acceptable carrier can include excipients, diluents, adjuvants, and vehicles, as well as implant carriers, and inert, non-toxic solid or liquid fillers, diluents, or encapsulating material that does not react with the active ingredients of the invention and do not themselves induce the production of antibodies harmful to the individual receiving the composition, and which may be administered without undue toxicity. Examples include, but are not limited to, phosphate buffered saline, physiological saline, water, and emulsions, such as oil / water emulsions. The carrier can be a solvent or dispersing medium containing, for example, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Example of suitable excipients include, but are not limited to, sorbitol, Tween80, andliquids such as water, saline, glycerol, and ethanol. Pharmaceutically acceptable salts can be included therein, for example, mineral acid salts such as hydrochlorides, hydrobromides, phosphates, sulfates, and the like; and the salts of organic acids such as acetates, propionates, malonates, benzoates, and the like. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, and the like, may be present in such vehicles. Formulations are described in a number of sources that are well known and readily available to those skilled in the art. For example, Remington’s Pharmaceutical Sciences (Martin EW

[1995] Easton Pennsylvania, Mack Publishing Company, 19thed.) describes formulations which can be used in connection with the subject invention. As appropriate compositions there may be cited all compositions usually employed for systemically or topically administering drugs. These pharmaceutical compositions are desirably in unitary dosage form suitable, preferably, for administration mucosally, nasally, orally, percutaneously, subcutaneously, or by parenteral injection. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs and solutions; or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets Because of their ease in administration, tablets and capsules often represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, though other ingredients, for example, to aid solubility, may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution.The term “expression level” as used herein refers to detecting the amount or level of expression of a biomarker of the present invention. The act of actually detecting the expression level of a biomarker refers to the act of actively determining whether a biomarker is expressed in a sample or not. This act can include determining whether the biomarker expression is upregulated, downregulated, or substantially unchanged as compared to a control level expressed in a sample. The expression level in some cases may refer to detecting transcription of the gene encoding a biomarker protein and / or to detecting translation of the biomarker protein.Expression of genes / transcripts and / or polypeptides encoded by the genes represented by the biomarkers of the present invention can be measured by any of avariety of methods known in the art. In general, expression of a nucleic acid molecule (e.g., RNA or DNA) can be detected by any suitable method or technique of measuring or detecting gene or polynucleotide sequence or expression. Such methods include, but are not limited to, polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR), in situ PCR, quantitative PCR (q-PCR), in situ hybridization, Southern blot, Northern blot, sequence analysis, microarray analysis, detection of a reporter gene, or any other DNA / RNA hybridization platforms.The term “quantifying” or “quantitating” when used in the context of quantifying transcription levels of a gene can refer to absolute or relative quantification. Absolute quantification can be achieved by including known concentration(s) of one or more target nucleic acids and referencing the hybridization intensity of unknowns with the known target nucleic acids (e.g., through the generation of a standard curve). Alternatively, relative quantification can be achieved by comparison of hybridization signals between two or more genes, or between two or more treatments to quantify the changes in hybridization intensity and, by implication transcription level.The term “gene expression product” or “expression product” as used herein refers to an RNA transcribed from a gene (either pre- or post-processing) or an amino acid (e.g. a polypeptide, protein, or peptide regardless of any secondary modifications, such as glycosylation, lipidation or phosphorylation) encoded by the gene and generated by the gene when the gene is transcribed (either pre- or post-modification) and translated. An agent is said to increase gene expression if the application of a therapeutically effective amount of the agent to a cell or subject results in an increase in either an RNA or polypeptide expression product or both. An agent is said to decrease gene expression if the application of a therapeutically effective amount of the agent to a cell or subject results in a decrease in either an RNA or polypeptide expression product or both.The term “polynucleotide” as used herein refers to a polymeric molecule that has a backbone that supports bases capable of hydrogen bonding to typical polynucleotides. The polymer backbone presents the bases in a manner that is effective to allow such hydrogen bonding in a sequence specific fashion between the polymeric molecule and a typical polynucleotide, such as single-stranded DNA. Polymeric molecules include both single and double stranded DNA or RNA and can include polymers having backbone modifications. It includes the recited sequences as well as their complementary sequences, which can be easily ascertained by those of ordinary skill in the art.The term “nucleic acid” as used herein may be double-stranded, single-stranded, or contain portions of both double and single stranded sequence. If the nucleic acid is single-stranded, the sequence of the other strand is also identifiable and thus the definition includes the complement of the sequence disclosed.The term “peptide” as used herein refers to short polymers formed from the linking, in a defined order, of a-amino acids. The link between one amino acid residue and the next is known as an amide bond or a peptide bond. Proteins are polypeptide molecules (or consist of multiple polypeptide subunits). The distinction is that peptides are short and polypeptides / proteins are long. There are several different conventions to determine these. Peptide chains that are short enough to be made synthetically from the constituent amino acids are called peptides, rather than proteins, with one commonly understood dividing line at about 50 amino acids in length.The term “polypeptide” as used herein refers to a compound made up of a singlechain of amino acid residues that are linked by peptide bonds. The term “protein” may be synonymous with the term “polypeptide” or may refer, in addition, to a complex of two or more polypeptides. Generally, polypeptides and proteins are formed predominantly of naturally occurring amino acids.An “isolated polynucleotide” as used herein refers to a polynucleotide which is separated from other nucleic acid molecules which are present in the natural source of the polynucleotide. Preferably, an “isolated polynucleotide” is free of sequences which naturally flank the polynucleotide in the genomic DNA of the organism from which the nucleic acid is derived. An “isolated polynucleotide” is substantially free of other cellular material, gel materials, and culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. The polynucleotides of the present invention may be isolated from a variety of sources, such as PCR amplification from genomic DNA, mRNA, or cDNA libraries derived from the mRNA using standard techniques The term “differential expression” as used herein refers to qualitative or quantitative differences in the temporal and / or spatial gene expression patterns within and among cells and tissues. A differentially expressed gene may qualitatively have its expression altered, including an activation or inactivation, such as in normal versus diseased tissue. Genes may be turned off or on in a given state relative to another state thus allowing comparison of two or more states. A qualitatively regulated gene may exhibit an expression pattern within a state or cell type that can be detectable by standard techniques. Alternatively, the difference in expression may be quantitativesuch that expression of the gene is modulated, up-regulated (resulting in an increased amount of transcript), or down-regulated (resulting in a decreased amount of transcript). The degree to which expression varies needs to be large enough to quantify via standard characterization techniques such as expression arrays, quantitative reverse transcriptase PCR, Northern blot analysis, real-time PCR, in situ hybridization, and RNase protection.The terms “overexpression” and “underexpression” as used herein refers to the expression of a gene of a patient at a greater or lesser level, respectively, than the normal or control expression of the gene, as measured by gene expression product expression such as mRNA or protein expression, in a sample that is greater than the standard of error of the assay used to assess the expression.Methods to measure protein / polypeptide expression levels of selected biomarkers in the present invention include, but are not limited to: Western blot, immunoblot, enzyme-linked immunosorbant assay (ELISA), radioimmunoassay (RIA), immunoprecipitation, surface plasmon resonance, chemiluminescence, fluorescent polarization, phosphorescence, immunohistochemical analysis, liquid chromatography mass spectrometry (LC-MS), matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF), mass spectrometry, microcytometry, microarray, microscopy, fluorescence activated cell sorting (FACS), flow cytometry, and assays based on a property of the protein including but not limited to DNA binding, ligand binding, or interaction with other protein partners.“Sample,” as used herein, refers to a composition that is obtained or derived from a subject and / or individual of interest that contains a cellular and / or other molecular entity that is to be characterized and / or identified, for example, based on physical, biochemical, chemical, and / or physiological characteristics. For example, the phrase “disease sample” and variations thereof refers to any sample obtained from a subject of interest that would be expected or is known to contain the cellular and / or molecular entity that is to be characterized. Samples include, but are not limited to, tissue samples, primary or cultured cells or cell lines, cell supernatants, cell lysates, platelets, serum, plasma, vitreous fluid, lymph fluid, synovial fluid, follicular fluid, seminal fluid, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, perspiration, mucus, tumor lysates, and tissue culture medium, tissue extracts such as homogenized tissue, tumor tissue, cellular extracts, and combinations thereof“Tissue sample” or “cell sample” is meant a collection of similar cells obtained from a tissue of a subject or individual. The source of the tissue or cell sample may be solid tissue as from a fresh, frozen, and / or preserved organ, tissue sample, biopsy, and / or aspirate; blood or any blood constituents such as plasma; bodily fluids such as cerebral spinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid; cells from any time in gestation or development of the subject. The tissue sample may also be primary or cultured cells or cell lines. Optionally, the tissue or cell sample is obtained from a disease tissue / organ. For instance, a “tumor sample” is a tissue sample obtained from a tumor or other cancerous tissue. The tissue sample may contain a mixed population of cell types (e.g. , tumor cells and non-tumor cells, cancerous cells, and non-cancerous cells). The tissue sample may contain compounds which are not naturally intermixed with the tissue in nature such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, or the likeA “tumor cell" as used herein, refers to any tumor cell present in a tumor or a sample thereof. Tumor cells may be distinguished from other cells that may be present in a tumor sample, for example, stromal cells and tumor-infiltrating immune cells, using methods known in the art and / or described herein.A “reference sample,” “reference cell,” “reference tissue,” “control sample,” “control cell,” or “control tissue,” as used herein, refers to a sample, cell, tissue, standard, or level that is used for comparison purposes In one embodiment, a reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and / or non-diseased part of the body (e.g , tissue or cells) of the same subject or individual. For example, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue may be healthy and / or non-diseased cells or tissue adjacent to the diseased cells or tissue (e.g., cells or tissue adjacent to a tumor). In another embodiment, a reference sample is obtained from an untreated tissue and / or cell of the body of the same subject or individual. In yet another embodiment, a reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and / or non-diseased part of the body (e.g., tissues or cells) of an individual who is not the subject or individual. In even another embodiment, a reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from an untreated tissue and / or cell of the body of an individual who is not the subject or individual. The term “biomarker” is used herein to refer to a molecule whose level of nucleic acid or protein product has a quantitatively differential concentration or level with respect to an aspect of a biological state of a subject. “Biomarker” is used interchangeablywith “marker” herein. The level of the biomarker can be measured at both the nucleic acid level as well as the polypeptide level. At the nucleic acid level, a nucleic acid gene or a transcript which is transcribed from any part of the subject’s chromosomal and extrachromosomal genome, including for example the mitochondrial genome, may be measured. Preferably an RNA transcript, more preferably an RNA transcript includes a primary transcript, a spliced transcript, an alternatively spliced transcript, or an mRNA of the biomarker is measured. At the polypeptide level, a pre-propeptide, a propeptide, a mature peptide or a secreted peptide of the biomarker may be measured. A biomarker can be used either solely or in conjunction with one or more other identified biomarkers so as to allow correlation to the biological state of interest as defined herein. Specific examples of biomarkers covered by the present invention include genes and gene expression products involved in collagen synthesis, more specifically generation of FACITs. More specifically, genes COL12A1, COL22A1, COL9A2, their associated gene products, and combinations thereof.The term “cell” or “cells” is used synonymously herein and refers to in vitro cultures of mammalian cells grown and maintained as known in the art, as well as biological samples obtained from tumor specimens or normal specimens in vivo.MethodsIn an aspect, a method of treating uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs) In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is premenopausal. The therapeutically effective amount of the composition acts to downregulate expression of a gene or gene product encoding the at least one FACIT in the uterine leiomyoma in the patient to treat the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In certain aspects, the at least one FACIT is collagen IX. In a preferable aspect, the at least one FACIT is collagen XII. In certain other aspects, the at least one FACIT is collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX, collagen XII, and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XII. In certain other aspects, the at least one FACIT is a combination of collagen XII and collagen XXII. In certain other aspects, the at least one FACIT is a combination ofcollagen IX and collagen XXII. In certain aspects, the at least one gene or gene product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof. In certain preferred aspects, the at least one gene or gene product that is downregulated is COL12A1. In certain other aspects, the at least one gene or gene product that is downregulated is COL9A2. In certain other aspects, the at least one gene or gene product that is downregulated is COL22A1. In certain other aspects, the combination of genes COL12A1, COL9A2, and COL22A1 are downregulated. In certain other aspects, the combination of COL12A1 and COL9A2 are downregulated. In certain other aspects, the combination of COL12A1 and COL22A1 are downregulated. In certain other aspects, the combination of COL9A2 and COL22A1 are downregulated.In aspects targeting COL12A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and combinations thereof. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the therapeutic agent is OMDI. In certain aspects, the therapeutic agent is OMD. In certain aspects, the therapeutic agent is caffeine. In certain aspects, the therapeutic agent is decitabine. In certain aspects, the therapeutic agent is U0126. In certain aspects, the therapeutic agent is palbociclib. In certain aspects, the therapeutic agent is geldanamycin. In certain aspects, the therapeutic agent is rifocin. In certain aspects, the therapeutic agent is eupachlorin. In certain aspects, the therapeutic agent is nanaomycin. In certain aspects, the therapeutic agent is nilotinib.In aspects targeting COL22A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL22A1, kaempferol, EMD-1204831, a CDK4 / 6 inhibitor such as palbociclib, a matrix metalloproteinase (MMP) inhibitor, an engrailed homeobox 1 (EN1) inhibitor, and combinations thereof. In certain aspects, the therapeutic agent is kaempferol. In certain aspects, the therapeutic agent is EMD-1204831. In certain aspects, the therapeutic agent is a combination of kaempferol and EMD-1204831. In certain aspects, the therapeutic agent is palbociclib.In aspects targeting COL9A2 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL9A2, SOX9 inhibitors, and combinations thereof.In an aspect, a method of treating uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. The therapeutically effective amount of the composition reduces the production / synthesis and / or amount of the at least one FACIT in the uterine leiomyoma in the patient to treat the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain aspects in which the FACIT is collagen type XII, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In an aspect, a method of inhibiting growth of uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition acts to downregulate expression of at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to inhibit growth of the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In certain aspects, the at least one FACIT is collagen IX. In a preferable aspect, the at least one FACIT is collagen XII. In certain other aspects, the at least one FACIT is collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX, collagen XII, and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XII. In certain other aspects, the at least one FACIT is a combination ofcollagen XII and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XXII In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1. In certain other aspects, the at least one gene or gene expression product that is downregulated is COL9A2. In certain other aspects, the at least one gene or gene expression product that is downregulated is COL22A1. In certain other aspects, the combination of genes COL12A1, COL9A2, and COL22A1 are downregulated. In certain other aspects, the combination of COL12A1 and COL9A2 are downregulated. In certain other aspects, the combination of COL12A1 and COL22A1 are downregulated. In certain other aspects, the combination of COL9A2 and COL22A1 are downregulated.In aspects targeting COL12A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and combinations thereof. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the therapeutic agent is OMDI. In certain aspects, the therapeutic agent is OMD. In certain aspects, the therapeutic agent is caffeine. In certain aspects, the therapeutic agent is decitabine. In certain aspects, the therapeutic agent is U0126. In certain aspects, the therapeutic agent is palbociclib. In certain aspects, the therapeutic agent is geldanamycin. In certain aspects, the therapeutic agent is rifocin. In certain aspects, the therapeutic agent is eupachlorin. In certain aspects, the therapeutic agent is nanaomycin. In certain aspects, the therapeutic agent is nilotinib.In aspects targeting COL22A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL22A1, kaempferol, EMD-1204831, a CDK4 / 6 inhibitor such as palbociclib, a matrix metalloproteinase (MMP) inhibitor, an engrailed homeobox 1 (EN1) inhibitor, and combinations thereof. In certain aspects, the therapeutic agent is kaempferol. In certain aspects, the therapeutic agent is EMD-1204831. In certain aspects, the therapeutic agent is a combination of kaempferol and EMD-1204831. In certain aspects, the therapeutic agent is palbociclib.In aspects targeting COL9A2 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL9A2, SOX9 inhibitors, and combinations thereof.In an aspect, a method of inhibiting growth of uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof The therapeutically effective amount of the composition reduces the production of or amount of the at least one FACIT in the uterine leiomyoma in the patient to inhibit growth of the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain aspects in which the FACIT is collagen type XII, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the patient is pre-menopausal. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In an aspect, a method of inhibiting tumor cell proliferation in uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition acts to downregulate expression of at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to inhibit tumor cell proliferation in the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In certain aspects, the at least one FACIT is collagen IX. In a preferable aspect, the atleast one FACIT is collagen XII. In certain other aspects, the at least one FACIT is collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX, collagen XII, and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XII. In certain other aspects, the at least one FACIT is a combination of collagen XII and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XXII. In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1. In certain other aspects, the at least one gene or gene expression product that is downregulated is COL9A2. In certain other aspects, the at least one gene or gene expression product that is downregulated is COL22A1. In certain other aspects, the combination of genes or gene expression products COL12A1, COL9A2, and COL22A1 are downregulated. In certain other aspects, the combination of COL12A1 and COL9A2 are downregulated. In certain other aspects, the combination of COL12A1 and COL22A1 are downregulated. In certain other aspects, the combination of COL9A2 and COL22A1 are downregulated.In aspects targeting COL12A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and combinations thereof. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the therapeutic agent is OMDI. In certain aspects, the therapeutic agent is OMD. In certain aspects, the therapeutic agent is caffeine. In certain aspects, the therapeutic agent is decitabine. In certain aspects, the therapeutic agent is U0126. In certain aspects, the therapeutic agent is palbociclib. In certain aspects, the therapeutic agent is geldanamycin. In certain aspects, the therapeutic agent is rifocin. In certain aspects, the therapeutic agent is eupachlorin. In certain aspects, the therapeutic agent is nanaomycin. In certain aspects, the therapeutic agent is nilotinib.In aspects targeting COL22A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL22A1, kaempferol, EMD-1204831, a CDK4 / 6 inhibitor such as palbociclib, a matrix metalloproteinase (MMP) inhibitor, an engrailed homeobox 1 (EN1) inhibitor, and combinations thereof. In certain aspects, the therapeutic agent is kaempferol. In certain aspects, thetherapeutic agent is EMD-1204831. In certain aspects, the therapeutic agent is a combination of kaempferol and EMD-1204831. In certain aspects, the therapeutic agent is palbociclib. In aspects targeting COL9A2 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL9A2, SOX9 inhibitors, and combinations thereof.In an aspect, a method of inhibiting tumor cell proliferation in uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production / synthesis of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. The therapeutically effective amount of the composition reduces the amount of the at least one FACIT in the uterine leiomyoma in the patient to inhibit tumor cell proliferation in the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain aspects in which the FACIT is collagen type XII, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the patient is pre-menopausal. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In an aspect, a method of reducing structural integrity and / or reducing stiffness of a uterine leiomyoma in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least gene or gene expression product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition acts to downregulate expression of the at least one gene or gene expression product encoding at least one FACIT in the uterine leiomyoma in the patient to reduce the structural integrity and / or reduce the stiffness of the uterineleiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In certain aspects, the at least one FACIT is collagen IX. In a preferable aspect, the at least one FACIT is collagen XII. In certain other aspects, the at least one FACIT is collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX, collagen XII, and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XII. In certain other aspects, the at least one FACIT is a combination of collagen XII and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XXII In certain aspects, the at least one gene or gene product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof. In certain preferred aspects, the at least one gene or gene product that is downregulated is COL12A1. In certain other aspects, the at least one gene or gene product that is downregulated is COL9A2. In certain other aspects, the at least one gene or gene product that is downregulated is COL22A1. In certain other aspects, the combination of COL12A1, COL9A2, and COL22A1 are downregulated. In certain other aspects, the combination of COL12A1 and COL9A2 are downregulated. In certain other aspects, the combination of COL12A1 and COL22A1 are downregulated. In certain other aspects, the combination of COL9A2 and COL22A1 are downregulated.In aspects targeting COL12A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and combinations thereof. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the therapeutic agent is OMDI. In certain aspects, the therapeutic agent is OMD. In certain aspects, the therapeutic agent is caffeine. In certain aspects, the therapeutic agent is decitabine. In certain aspects, the therapeutic agent is U0126. In certain aspects, the therapeutic agent is palbociclib. In certain aspects, the therapeutic agent is geldanamycin. In certain aspects, the therapeutic agent is rifocin. In certain aspects, the therapeutic agent is eupachlorin. In certain aspects, the therapeutic agent is nanaomycin. In certain aspects, the therapeutic agent is nilotinib.In aspects targeting COL22A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL22A1, kaempferol, EMD-1204831, a CDK4 / 6 inhibitor such as palbociclib, a matrix metalloproteinase (MMP) inhibitor, an engrailed homeobox 1 (EN1) inhibitor, and combinations thereof. Incertain aspects, the therapeutic agent is kaempferol. In certain aspects, the therapeutic agent is EMD-1204831. In certain aspects, the therapeutic agent is a combination of kaempferol and EMD-1204831. In certain aspects, the therapeutic agent is palbociclib. In aspects targeting COL9A2 downregulation, the therapeutic agent may be selected from RNA interference agent targeting COL9A2, SOX9 inhibitors, and combinations thereof.In an aspect, a method of reducing structural integrity and / or reducing stiffness in uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. The therapeutically effective amount of the composition reduces the structural integrity and / or stiffness of the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain aspects in which the FACIT is collagen type XII, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.In an aspect, a method of preventing fibrosis in uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a composition, the composition comprising a therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least gene or gene expression product encoding at least one fibril-associated collagen with interrupted triple helices (FACITs). In certain aspects, the composition further comprises a pharmaceutically acceptable carrier. In certain aspects, the patient is pre-menopausal. The therapeutically effective amount of the composition acts to downregulate expression of at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to preventfibrosis in the uterine leiomyoma. In certain aspects, the at least one FACIT is collagen IX, collagen XII, collagen XXII, or combinations thereof. In certain aspects, the at least one FACIT is collagen IX. In a preferable aspect, the at least one FACIT is collagen XII. In certain other aspects, the at least one FACIT is collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX, collagen XII, and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XII. In certain other aspects, the at least one FACIT is a combination of collagen XII and collagen XXII. In certain other aspects, the at least one FACIT is a combination of collagen IX and collagen XXII. In certain aspects, the at least one gene or gene expression product being downregulated is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof. In certain preferred aspects, the at least one gene or gene expression product that is downregulated is COL12A1. In certain other aspects, the at least one gene or gene expression product that is downregulated is COL9A2 In certain other aspects, the at least one gene or gene expression product that is downregulated is COL22A1. In certain other aspects, the combination of COL12A1, COL9A2, and COL22A1 are downregulated. In certain other aspects, the combination of COL12A1 and COL9A2 are downregulated. In certain other aspects, the combination of COL12A1 and COL22A1 are downregulated. In certain other aspects, the combination of COL9A2 and COL22A1 are downregulated.In aspects targeting COL12A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, nilotinib, and combinations thereof. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the therapeutic agent is OMDI. In certain aspects, the therapeutic agent is OMD. In certain aspects, the therapeutic agent is caffeine. In certain aspects, the therapeutic agent is decitabine. In certain aspects, the therapeutic agent is U0126. In certain aspects, the therapeutic agent is palbociclib. In certain aspects, the therapeutic agent is geldanamycin. In certain aspects, the therapeutic agent is rifocin. In certain aspects, the therapeutic agent is eupachlorin. In certain aspects, the therapeutic agent is nanaomycin. In certain aspects, the therapeutic agent is nilotinib.In aspects targeting COL22A1 downregulation, the therapeutic agent may be selected from an RNA interference agent targeting COL22A1, kaempferol, EMD-1204831, a CDK4 / 6 inhibitor such as palbociclib, a matrix metalloproteinase (MMP)inhibitor, an engrailed homeobox 1 (EN1) inhibitor, and combinations thereof. In certain aspects, the therapeutic agent is kaempferol. In certain aspects, the therapeutic agent is EMD-1204831. In certain aspects, the therapeutic agent is a combination of kaempferol and EMD-1204831. In certain aspects, the therapeutic agent is palbociclib. In aspects targeting COL.9A2 downregulation, the therapeutic agent may be selected from RNA interference agent targeting COL9A2, SOX9 inhibitors, and combinations thereof.In an aspect, a method of preventing fibrosis in uterine leiomyomas in a patient in need thereof is disclosed comprising administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing the production of or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT). In certain aspects, the at least one FACIT comprises collagen type XII, collagen type IX, collagen type XXII, and combinations thereof. The therapeutically effective amount of the composition prevents fibrosis in the uterine leiomyoma. In certain aspects, the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof. In certain preferred aspects, the FACIT is collagen type XII. In certain aspects in which the FACIT is collagen type XII, the therapeutic agent is an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib. In certain aspects, the therapeutic agent is the shRNA of SEQ ID NO: 1. In certain aspects, the patient is pre-menopausal. In certain aspects, a pharmaceutical composition comprised of the therapeutic agent, and a pharmaceutically acceptable carrier is administered to the patient.ExamplesUterine leiomyomas are benign tumors founds in as many as 77% of reproductive-aged women. Despite the fact that leiomyomas are a frequent cause of pelvic pain, vaginal bleeding and infertility, medical options for managing these common tumors remain limited. Abundant deposition of extracellular matrix, most notably Types I and III collagen, is a well-established hallmark of uterine leiomyomas. In contrast to healthy myometrium, organization of the extracellular matrix (ECM) in leiomyomas is characterized by an irregular alignment and loose packing that increases tissue stiffness and intratumoral shear stress and plays a key role in driving the growth of these neoplasms. Although fibrosis is a well-established hallmark of leiomyomas, themechanisms by which the unique ECM found in leiomyoma promotes growth of these tumors remain poorly understood.The inventors found that overexpression of COL12A1 and multiple other collagens (COL9A2 and COL22A1) with interrupted triple helices (FACITs) are a robust feature of uterine leiomyomas. Interrogation of transcripts generated by single cell profiling indicates that COL12A1 is robustly expressed in diverse cell types in leiomyomas and mediates communication between uterine myocytes, myofibroblasts, macrophages, myeloid cells, and endothelium. In primary cultures derived from human leiomyoma, functional silencing of COL12A1 reduces proliferation, promotes G1-S cell cycle arrest, and inhibits AKT1 -mediated signaling. Targeting COL12A1 expression disrupts the morphological compaction and survival of three-dimensional leiomyoma organoids promoted by mechanical stress and dramatically alters the abundance and histologic features of organoid cell populations Collectively, these findings establish COL12A1 as a novel, dual function structural and signaling mediator that integrates ECM remodeling and mechanical stress to drive leiomyoma growth and identify a promising new target for treatment.COL12A1 gene expression patterns were evaluated using a previously validated data set (Delaney, et al., 2017). In addition, patterns of COL12A1 expression in tissues and primary cultured cells were validated using semi-quantitative real-time PCR (RT-qPCR) and Western blot. Transcript-specific siRNAs were used to target COL12A1 expression in vitro. CellTiter 96® AQueous One Solution Cell Proliferation Assay (Promega) was used to evaluate proliferation; flow cytometry was used to evaluate cell cycle distribution Western blot was used for detecting the P-Akt and CylinDI signaling pathway. Statistical significance was assessed using paired Student’s t-tests assuming normal distribution of data. The inventors found that COL12A1 was robustly overexpressed in specimens of leiomyomas collected during the proliferative (2.5- fold, n=9, p=0.00026) and secretory (2-fold, n=10, p=0.012) menstrual phase. Using a separate set of specimens, COL12A1 was also overexpressed in proliferative (n=6, p=0.033) and secretory (n=6, p=0.015) leiomyomas, but not in postmenopausal tissues (n=6, p=0.18). COL12A1 was also significantly increased in leiomyoma primary cultured cells compared to matched myometrium primary cells (n=9, p=0.04). Knockdown of COL12A1 in primary cultures derived from leiomyomas slowed proliferation at 48 and 72 hours (n=3), increased the proportion of myocytes in G1 (p=0004) and decreased in proportion of S (p=0.006) and G2 (p=0.03) when compared to cultures transfected with a non-targeting scrambled control (n=4).C0L12A1 knockdown was accompanied by decreased expression of phospho-Akt and CyclinDlExample 1 - Dysregulated FACIT expression is a robust feature of uterine leiomyomasThe inventors evaluated the expression of known FACIT collagens (Types IX, XII, XIV, XVI, XIX, XX, XXI, and XXII) in matched specimens of leiomyoma and healthy myometrium. As shown in FIG. 1A, robust expression of multiple FACIT collagens, specifically COL12A1, COL14A1, and COL16A1, was found in both myometrium and leiomyoma while only minimal expression of transcripts for COL9A1, COL9A2, COL9A3, COL20A1 or COL21A1 was observed.T ranscripts for multiple FACIT collagens were expressed at significantly higher levels in leiomyomas independent of menstrual phase. These included COL12A1 (Proliferative phase myo vs leio: 1843 ± 170 vs 3990 ± 444, n=12, p= 0.0004; Secretory phase myo vs leio: 2241 ± 331 vs 4916 ± 846, n=9, p=0.007) as well as COL22A1 and COL9A2. (FIG. 1A) Although levels of FACIT expression did not appear to be associated with menstrual phase, lower levels of COL9A1 transcript were observed in specimens of myometrium and leiomyomas collected during the secretory menstrual phase, suggesting that its expression may be selectively regulated by steroid hormone exposure.To confirm these observations, the inventors interrogated total RNA prepared from additional tissue specimens by semi-quantitative RT-qPCR. These data confirmed that COL12A1 was expressed at significantly higher in leiomyomas, regardless of whether COL12A1 transcript was measured by RT-qPCR (Proliferative myo: 113.2 ± 7.96 ng / ml vs proliferative leio:1334 ± 4.64 ng / ml ; Secretory myo: 81.95 ± 7.69 ng / ml vs secretory leio: 110.6 ± 469 ng / ml; n=6 each, p= 0.01) or ELISA, independent of menstrual phase. (FIG. 1C). No difference in levels of COL12A1 expression were observed when transcript levels were evaluated by RT-qPCR in matched specimens of leiomyoma and myometrium collected from menopausal subjects (FIG. 1D). Using immunohistochemistry, the inventors found that myometrium exhibited weaker and more sporadic COL12A1 immunoreactivity, whereas leiomyomas were characterized by much more intense staining diffusely throughout each tumor. (FIG.1B) Robust COL12A1 expression was also noted at the interface between the leiomyoma and surrounding myometrial tissue (FIG. 1B). It was semi-quantitatively confirmed that COL12A1 immunoreactivity was significantly greater in leiomyomasthan adjacent healthy myometrium in all subjects evaluated (H-SCORE: 136.2 ± 2504 vs 75.67 ± 21.40, p<0.0001, n=15 sections from 5 subjects). (FIG. 1 C) To determine whether COL12A1 expression was a durable, feature of leiomyomas that persists in vitro, the inventors generated primary cultures from matched specimens of myometrium and leiomyomas. COL12A1 was expressed at significantly greater levels in primary cultures derived from leiomyomas than those derived from myometrium (fold change myo vs leio; 2.7 ± 2.9 vs 5.5 ± 60, n=6, p=0.04). (FIG. 1D).Greater COL12A1 expression was also observed when protein homogenates prepared from cell pellets were tested by Western blot (n=4, p>0.05). (FIG. 1D), and when ELISA was used to assess media bathing each culture (Myo: 16.0 ± 3 1 ng / ml vs 20.6 ± 2.1 ng / ml, n=6, p=0.01). (FIG. 1E).As noted herein, the inventors found that COL12A1, COL22A1 and COL9A2 are highly overexpressed in uterine leiomyomas while COL9A1 is downregulated. Furthermore, only minimal expression of COL21A1 or COL22A1 is observed in healthy myometrium (FIG. 1). In part, this pattern of expression may reflect the fact that all fibrillar collagens are extracellular matrix of leiomyomas is not uniformly most abundantly expressed in leiomyomas and the organization of these collagens required to support tumor growth. Unlike classical fibrillar collagens, FACIT collagens do not form fibrils, but instead interact fibrillar collagens to stabilize ECM architecture and resilience. In other cells and tissues, such as cartilage, COL12A1 directly bind and decorate the surface of Type I collagen, where it plays a key role in their organization and interaction with other components of the extracellular matrix. Notably abundant in tissues subjected to high tensile stress, such as tendons and bones, COL12A1 contributes to mechanical strength of joints and other tissues subjected to mechanical stress. The data indicate that the high levels of COL12A1 that characterize leiomyomas play a key role in promoting the activity of AKT1-mediated signaling pathways to promote cell proliferation.Example 2 - Distinct cell subtypes drive COL12A1 expression and collagen signaling in leiomyomasTo gain deeper insight into COL12A1-linked cell subtypes and signaling networks within leiomyomas, the inventors analyzed profiles generated from matched specimens of healthy myometrium and leiomyoma profiled by single-cell sequencing (GSE162122). The goal was to decipher the COL12A1 expression at the single-cell level and understand potential patterns of communication between distinct subsets of cells in the leiomyoma complex. After stringent quality control and datanormalization, the inventors gained transcriptomes for a total of 11,354 cells from leiomyomas and 11,354 cells from their normal counterparts. PCA (Principal Component Analysis) and UMAP (Uniform Manifold Approximation Projection) determined 27 different clusters for both leiomyomas and myometrium. 12 major cell types were determined by using the cell markers shown in Table 1 with semisupervised category identification and assignment (SCINA) which was used to perform cell annotation, including smooth muscle cells, smooth muscle growing, tumor muscle, fibroblasts, myofibroblasts, T cells, B cells, NK cells, epithelial cells, myeloid cells, macrophages, and endothelial cells. Using Seurat RidgePlot, the inventors found that only a small proportion of myofibroblasts, fibroblasts or smooth muscle expressed COL12A1 in healthy myometrium (FIG. 2A). However, in leiomyomas, robust COL12A1 expression was not only observed in myofibroblasts and different subtypes of smooth muscle cells, but also multiple types of inflammatory, epithelial, and endothelial cells (FIG.2B)T o further characterize the collagen network, the inventors performed CellChat on the transcriptomics data by integrated CellChatDB with CellPhoneDB. Both were used because the CellChat (v2) can refer to cell-state specific ligand-receptor interactions with a given comprehensive signal molecule interaction reference database (DB), while the CellPhone (v5) contains more comprehensive information on collagen networks. When CellChat was used to evaluate the number and directionality of signals between cell subpopulations, the inventors found that fibroblasts serve as the primary output for collagen-mediated signaling in healthy myometrium, communicating with myofibroblasts and multiple subtypes of smooth muscle cells (FIG. 2C). In contrast, collagen-mediated signals in leiomyomas were much more numerous and involved a greater number of different cell types. As shown in FIG.2D, myofibroblasts rather than fibroblasts serve as primary source of outgoing collagen-mediated signals in leiomyomas, largely directed at endothelial cells and multiple subtypes of smooth muscle. Outgoing collagen-mediated signals directed at other cells, including macrophages, myeloid and endothelial cells, were also identified. These latter patterns readily differentiated leiomyomas from health myometrium, where collagen-mediated signaling that involved inflammatory cells were infrequent. Notably, NK cells in leiomyomas were found to direct outgoing signals towards endothelial cells and multiple types of smooth muscle; similar signals were not detected in healthy myometrium (FIG. 2C, D). Activity of NK cells in leiomyomas contrasted with the limited number of collagen-mediated signals found to arise from B- or T-lymphocytes, despite robust COL12A1 expression in both cell types. Nocollagen-mediated signals involving B-cells, T-cells or macrophages were identified in healthy myometrium.Overexpression of COL12A1 has been recently identified as a biomarker for biologically active fibroblasts. However, the analysis of single-cell transcriptomic data indicates that high levels of COL12A1 expression in fibroblasts to include multiple subtypes of uterine smooth muscles and myofibroblast populations. Furthermore, an analysis of receptor-donor dyads using CellChat further indicates that collagen mediated signaling plays a key role in mediating patterns of communication between myofibroblasts, fibroblasts, endothelial cells, and multiple types of inflammatory cells (FIG. 2B, D).Example 3 - COL12A1 regulates the proliferation of uterine myocytes Targeting the high levels of COL12A1 expression observed in primary cultures of leiomyomas consistently decreased proliferation when compared to controls transfected with a scrambled, non-targeting siRNA (n=3, p<0.05) (FIG. 3A-C) Decreased rates of proliferation induced by COL12A1 knockdown were accompanied by a greater proportion of cells accumulating at the G1 / S cell cycle checkpoint (FIG.3D-F; p<0.05). Evaluation of these cultures by RT-qPCR (Fig. 3G, I, K) and ELISA (Fig. 3H, J, L), confirmed a significant decrease in both COL12A1 mRNA transcripts and protein levels in response to transfection with COL12A1 -targeting siRNAs. Example 4 - COL12A1 is essential for the structural integrity and survival of leiomyoma organoidsThe inventors next targeted COL12A1 in a three-dimensional organoid model of leiomyomas to assess its role in the assembly and three-dimensional organization of these tumors Organoids generated from primary leiomyoma cultures transfected with shCOL12A1 (SEQ ID NO: 1) demonstrated less compact and more disperse morphology as evidenced by increased surface area when compared to controls generated from aliquots transfected with a scrambled non-targeting shRNA (n=3).(FIG. 4A-B) Impressively, this difference was readily observable within 24 hours of seeding media with dispersed cells. With continued incubation, shCOL12A1-transfected organoids became increasingly disorganized and fragile, demonstrating irregular, frayed borders that were easily disrupted even by gentle motion of the bathing media. In contrast, control cultures became progressively more compact and remained resilient to motion over the 2-week timeframe over which they were evaluated. Interestingly, disorganization of shCOL12A1-transfected organoidspersisted and worsened despite a return of COL12A1 expression towards baseline that began as early as 5 days after shCOL12A1 transfection (FIG.4C-F).Collectively, these data suggest that COL12A1 plays an essential role in orchestrating the composition, organization and compaction of leiomyoma organoids and supports a crucial role for COL12A1 in leiomyoma pathogenesis.Example 5 - COL12A1 mediates the impact of mechanical stress on myometrial organoidsGiven its key role in stabilizing cartilage in response to shear stress, the inventors believe that COL12A1 mediates the impact of mechanical stress on leiomyoma growth. While numerous distinct technical platforms have been used previously to study the impact of shear stress on uterine myocytes in vitro, the majority of these approaches cannot be readily adapted for use in studying a three-dimensional model system. The inventors used microgravity as a useful tool for understanding the role of mechanical stress in the growth and organization of uterine leiomyomas as simulated microgravity reduces cytoskeletal tension and triggers cellular remodeling. As such, simulated microgravity was used as a tool to apply mechanical stress globally and evenly to leiomyoma organoids.In tissues such as cartilage, COLA121 and other FACIT collagens tether components of extracellular matrix, playing an important role in their organization and stability. Given these properties, the inventors believed that COL12A1 expression in leiomyomas plays an important role in generating a unique fibrotic microenvironment important for their organization and growth. Simulated microgravity was used as a tool to apply mechanical stress globally and evenly to leiomyoma organoids. Cell suspensions derived from leiomyomas self-assembled into organoids much more rapidly when exposed to microgravity (FIG. 5A). Impressively, this impact was readily detectable as early as 24 hours following microgravity exposure.Prior work has demonstrated that the acute application of shear stress to dissociated fibroblasts rapidly induced COL12A1 expression which resolves following stimulus withdrawal. In light of this observation, the inventors show that the mechanical unloading resulting from microgravity also regulates COL12A1 expression in uterine myocytes. COL12A1 expression in primary myometrial cultures exposed to microgravity was evaluated. When compared to untreated, matched controls, transcripts for COL12A1 were found to be expressed at significantly higher levels in response to microgravity exposure, (fold change MG vs control: 1.75 ± 0.25 vs. 1.0 ±0.03 vs, n=4, p<0.0001). (FIG. 5D). To determine whether the overexpression of COL12A1 induced by mechanical stress was transcriptionally mediated, a reporter vector was used in which luciferase expression is regulated by COL12A1 promoter. As shown in FIG. 5B, microgravity exposure increased luciferase activity of primary myometrial cell cultures transfected with this vector but had little or no impact on control cultures transfected with empty vector or luciferase regulated by a minimal promoter (minP) element. These results confirmed that the observed increase in luciferase activity was specific to the consensus COL12A1 promoter (FIG. 5B) Application of mechanical stress results in increased luciferase activity in primary cultures of healthy myometrium (Myo#4, Myo #106, Myo#8) transfected with transfected with reporter vector in which luciferase activity is regulated by the consensus promoter for COL12A1. In contrast, mechanical stress had no impact on luciferase activity in cultures transected with either empty vector (Vector) lacking the coding sequence for luciferase or a second control in which luciferase transcription is regulated by minimal promoter element (MinP) (FIG. 5E) Mechanical stress had no impact on the compaction of organoids generated from primary leiomyoma cultures transfected with shCOL121. *p<0.05; **p<0.01. (FIG. 5F, G)These results establish COL12A1 as a mechanosensitive extracellular matrix component in the myometrium and provide further evidence of its role as a dynamic regulator of matrix remodeling under mechanical stress.Example 6 - Targeting COL12A1 and mechanical stress regulate transcription and reciprocally impact AKT1-mediated signalingTo decipher signaling pathways by which COL12A1 promotes proliferation in leiomyomas, primary leiomyoma cultures were transfected with either an siRNA targeting COL12A1 or a scrambled, non-targeting control. PI3K-Akt signaling pathway was examined via phosphorylated Akt (P-Akt) and Cyclin D1 protein levels in primary cell cultures. In leiomyoma cell cultures, COL12A1 knockdown (siCOL12A1) decreased Akt and Cyclin D1 protein levels compared to control (n=3).(FIG. 6A, B). Primary cultures derived from myometrium were exposed to microgravity for 24 hours, after levels of AKT 1 phosphorylation and cyclin D expression were evaluated. Results of these experiments indicated that expression of both phospho-AKT1 and Cyclin D1 protein were more abundant than matched controls (FIG. 6D, n=3). Quantitative analysis showed significant increase in mean band intensity in both P-Akt and Cyclin D1 after microgravity exposure (FIG. 6C, p<0.05, and p<0.01, respectively). These findings indicate that COL12A1 mediatesits effect via PI3K-Akt signaling in leiomyoma and mechanical stress activates this signaling pathway in myometrium. Thus, providing a mechanistic rationale for cellular response to biomechanical stimuliExample 7 - Treatment of leiomyoma (prophetic)A 25 year old female presents with heavy and pl bleeding and sever pelvic pain. Imaging is performed and the patient is diagnosed with leiomyomas. The patient is administered a therapeutically effective amount of a gene therapy comprising an RNA interference agent targeting COL12A1 for a therapeutically effective amount of time. The patient’s symptoms are reduced.Materials and Methods for all ExamplesHuman Tissue SpecimensAfter obtaining permission from the local Institutional Review Board, matched, flash frozen specimens of uterine leiomyoma and adjacent myometrium were obtained from the Tampa General Hospital / University of South Florida Health Precision Medicine Biorepository (PMBioR; RRID:SCR_027501). All specimens were collected from pre-menopausal subjects undergoing hysterectomy for benign indications; menstrual phase was established using self-reported dates provided by each subject for her most recent menses. Menstrual dating was histologically confirmed using established criteria as previously described in Guo, X. et al. (Spatially restricted ecto-5’-nucleotidase expression promotes the growth of uterine leiomyomas by modulating Akt activity, FASEB J., October 15, 2024; 38( 19): e70084), herein incorporated in its entirety. Myometrial specimens were collected at least 2 cm from the nearest leiomyoma. Only specimens from mid-proliferative (cycle day 10-14) or mid-secretory (cycle day 22-26) subjects were utilized. Any specimens collected from subjects actively treated with oral contraceptives or steroid hormones within 30 days of surgery were excluded. Written consent providing permission to share specimens and generate primary cell lines was available for each subject.RNA Isolation and Real Time Quantitative PCR (RT-qPCR)Total RNA was prepared from flash frozen specimens using the mirVana™ miRNA Isolation Kit (Invitrogen™, Waltham, MA). qScript cDNA SuperMix (Quantabio, Beverly, MA). After reverse transcription, transcript levels were quantified by semi-quantitative real-time PCR (RT-qPCR) using the following TaqMan gene expression assays (Applied Biosystems, Foster City, CA): Col12A1: Hs00189184_m1; Col9A2:Hs00895570_m1; Col22A1: Hs01377218_m1; and s18: Hs99999901_s1 All CT values obtained by were normalized to relative expression of 18S mRNA, as previously described in Guo, X. et al. (Spatially restricted ecto-5'-nucleotidase expression promotes the growth of uterine leiomyomas by modulating Akt activity, FASEB J., October 15, 2024; 38(19):e70084), herein incorporated by reference in its entirety. The AACT method was used to calculate relative fold-change in mean levels of gene expression as described in Livak, K.J and Schmitten, T. D. (Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method, Methods, December 2001; 25(4):402-8), incorporated herein in its entirety. All assays were run in triplicate.Western BlotProtein extracts were prepared using a standard lysis buffer supplemented with protease inhibitor (ThermoFisher, Cat #J64963.LQ) as previously described in Rosado, R. et al. ( Overexpression of high affinity Type I adenosine receptors promotes the growth of uterine leiomyomas, Mol Hum Reprod., July 3, 2025; 31(3):gaaf025) and Lee, J.H., et al. (Long-term outcomes of catheter-directed sclerotherapy for ovarian endometria, Diagn Interv Radiol., April 28, 2025; 31(3):249-252), incorporated in their entirety herein by reference. Validated primary antibodies for Col12A1 (1:1000 dilution; Abeam, Waltham, MA; RRID:AB_11140541), Phospho-Akt (1:1,000 dilution; Cell Signaling Technology Cat#4060, RRID:AB_2315049), Akt (1:1000 dilution; Cell Signaling Technology Cat# 4691, RRID:AB_915783), Cyclin D1 (1:1000 dilution; Cell Signaling Technology Cat# 55506, RRID:AB_2827374); GAPDH (1:2,000 dilution; Sigma-Millipore #G9545; RRID:AB_796208) were diluted in Tris-buffered saline supplemented with 0.1% Tween-20 (TBS-T) and 2% non-fat dried milk. After incubating blots overnight at 4°C, horseradish peroxidase (HRP)-conjugated anti-rabbit IgG antibody (1:3,000; Cell Signaling #7074S; RRID:AB_2099233) and HRP-conjugated anti-mouse IgG antibody (ThermoFisher# 31430, RRID: AB_228307) were used to visualize immunoreactivity by chemiluminescence using an Odyssey imaging system (LI-COR, Lincoln, NE). Band intensities were quantified by densitometry and normalized to GAPDH expression using Image STUDIO Lite Ver 5.2 (LI-COR; RRID:SCR_013715).Enzyme-linked immunosorbent assay (ELISA)COL12A1 concentration in tissues and cell culture supernatants was measured using MyBioSource Elisa kit (Cat# MBS076336, MyBioSource, Inc. San Diego, CA) according to the manufacturer’s instructions. Specimens were collected 48 hoursafter siRNA transfection, after which, equal masses of myometrium and leiomyoma cells were sonicated in 500 pl PBS, centrifuged at 10,000g for 10 minutes at 4°C and assayed immediately. Fluorescence at 450 nM was measured using a Cytation 5 Cell Imaging Multimode Reader (BioTek, Inc, Zeeland, Ml; RRID:SCR_019732).ImmunohistochemistryCOL12A1 expression was visualized in cross-sections from formalin-fixed, paraffin-embedded specimens of human leiomyomas as described (PMID: 39354726). In brief, COL12A1-specific immunoreactivity was visualized using a rabbit anti-COL12A1 (ab121304, Abeam, Inc; RRID:AB_11140541) diluted 1:75 in TBS-T supplemented 2% goat serum and visualized using the Vectastain Elite ABC-Peroxidase kit (Cat# PK-6100; Vector Labs, Inc ; RRID:AB_2336819) according to the manufacturer’s instructions. Slides were counterstained with 1:4 dilution of hematoxylin for 1 minute followed by bluing reagent for 1 minute, after which, they were dehydrated, cleared, and mounted with Entellan™ (Sigma-Aldrich). For negative controls, non-specific rabbit IgG (Cell Signaling Technology, #2729; RRID: AB_1031062) was used at a 1:75 dilution. COL12A1 expression was sequentially scored for both intensity and frequency in three randomly selected regions of interest at 20* magnification by two independent investigators (BU, SG) blinded to tissue type. Results are reported as a semi-quantitative histologic score (H-SCORE) as described in Delaney, M.A. et al. (A Role for Progesterone-Regulated sFRP4 Expression in Uterine Leiomyomas, J Clin Endocrinol Metab. 2017 Sep 1 ;102(9):3316-3326), incorporated herein in its entirety by reference. An average score from both investigators is reported for all specimens.In Vitro AssaysPrimary cultures were derived from matched specimens of heathy myometrium and leiomyoma and transfected with either 20 nM of human COL12A1 siRNA (Dharmacon, Inc., Lafayette, CO; Dharmacon Cat. J-011834-06-005) or ON-TARGETplus non-targeting control pool as previously described in Guo, X. et al. (Spatially restricted ecto-5’-nucleotidase expression promotes the growth of uterine leiomyomas by modulating Akt activity, FASEB J., October 15, 2024; 38(19):e70084), herein incorporated by reference in its entirety. In brief, fresh tissue specimens were minced and enzymatically digested at 37C overnight in a rotating shaker. The resulting cell slurries were filtered and, after removing debris, plated in culture media. After 24 hours, 2500 cells / well were seeded into 96 well plates in media supplemented with 25% FBS. CellTiter 96® AQueous One assay (RRID: G3582;Promega, Inc., Madison, Wl) was used to assess proliferation at 24, 48 and 72 hours. To confirm targeting efficacy, cell pellets were collected and used to prepare total RNA 24 hours following transfection. Protein extraction was performed 72 hours after transfection.To assess cell cycle distribution, 3 x 105cells were seeded into 60 mm dishes and incubated for 36 hours in media supplemented with 0.4% BSA. Cells were transfected with either 20 nM of human COL12A1 siRNA or ON-TARGETplus non-targeting control pool using Lipofectamine RNAiMax reagent. After 48 hours, cells were collected and washed twice with phosphate-buffered saline (PBS), after which, pelleted cells were fixed in 70% ethanol overnight, centrifuged at 1500g for 8 min, washed once with PBS, resuspended in 500pl FxCycle™ PI / RNAse Solution (Invitrogen), and incubated at room temperature in the dark for 30 minutes. BD FACS Canto II flow cytometer (Becton Dickinson, San Jose, CA) was used to sort labeled cells FlowJo software (v10.80; RRID: SCR_008520) was utilized to analyze DNA content.Organoid CultureLentivirus shCOL12A1 (SEQ ID NO: 1) was generated and used to infect the primary cultures as described in Sun, M. et al. (Collagen XII Regulates Corneal Stromal Structure by Modulating Transforming Growth Factor-p Activity. Am J Pathol. 2022 Feb;192(2):308-319), incorporated herein by reference in its entirety. In brief, HEK293T producer cells were transfected with the designated transfer plasmid containing sequences targeting COL12A1 and helper plasmids (encoding packaging, envelope, and Rev proteins). Supernatant was collected 72 hours after transfection, and directly incubated with target cultures, After incubating cultures with lentivirus-containing supernatants at 37°C for 24 hours, treated cells were seeded into 96-well ultra-low attachment plates (Corning) containing mesenchymal stem cell growth medium (MSCGM; Lonza, Walkersville, MD). All cultures were maintained in a 5% CO atmosphere at 37°C (PMID: 29381777). Organoids were evaluated at specified timepoints (24 hours and then 5, 10, 15 and 20 days by light microscopy (Keyence) at 4x magnification. Imaged (RRID:SCR_003070) was used for calculating spheroid area For 4',6-diamidino-2-phenylindole (DAPI) staining, organoids were incubated for 30 minutes at room temperature (RT) after adding 100 uL of 4% buffered formalin (ThermoFisher, J19943-K2) to each well. After washing twice with phosphate-buffered saline (PBS), organoids permeabilized with 02% Triton X-100 / PBS for 15 minutes at RT and stained with DAPI (1 pg / ml) for 1 hour in the dark. Spheroids were imaged at 4X magnification.Simulated MicrogravityAfter mounting culture plates at the center of the clinostat, specimens were rotated along axis of a commercially available clinostat system for 24 hours at speeds, according to the manufacturer’s instructions (Yuri GmbH, Meckebeuren, Germany), designed to average the gravitational vector and simulate microgravity at 0.01g. Throughout these experiments, cultures were maintained at 37°C with 5% CO2. For all experiments, matched controls were cultured under identical environmental conditions without being mounted within the clinostat.Reporter AssaysTo assay the impact of mechanical stress on transcriptional activity regulated by the COL12A1 promoter, the consensus promoter for COL12A1 was cloned into a commercially available expression vector, pGL4.23[luc2 / COL12A1] driving luciferase expression (Dual-Gio® Luciferase Reporter Assay System (Promega, Inc.) as described in Chiquet, M. et al. (The chick and human collagen alphal(XII) gene promoter-activity of highly conserved regions around the first exon and in the first intron. Eur J Biochem. 1998 Oct 15;257(2):362-71), herein incorporated in its entirety. After seeding 2.5 x 105cells into 6-well plates, cultures were transfected with either 2.5 pg of pGL4.23[luc2 / COL12A1], 25 pg of a control vector in which luciferase expression was regulated by a minimal functional promotor (pGL4.23[luc2 / minP]; Cat#E8411, Promega, Inc) or a negative control containing the COL12A1 promoter but lacking the sequences encoding Firefly luciferase. To assess transfection efficiency, all mixes were spiked with 2.5 pg of pGL4.75[hRluc / CMV] (Promega, Cat#E693, Promega, Inc; RRID:ADDGENE_24348). After 24 hours, cells were reseeded in 96-well plates and subjected to simulated microgravity for specific durations. Luciferase activity in cell lysates was measured using the BioT ek Cytation 5 Cell Imaging system.Analysis of Single cell Sequencing Data (scNGS)Publicly available data documenting the results of single cell Next Generation Sequencing (scNGS) of two specimens of human leiomyoma (GSM4942396, GSM492399) along with matched specimens of adjacent healthy myometrium (GSM5023321, GSM6509142) were selected for analysis. [PMID: 36001050] Raw reads (FASTQ files for each specimen were retrieved from the Sequence Read Archive (SRA) using the SRA toolkit version 3.1.1 (RRID:SCR_024350) and processed using Cell Ranger version 8.0.1 (10X Genomics, Pleasanton, CA) usingrefdata-gex-GRCh38-2020-A (GRCh38 / hg38) as the reference human genome) were obtained The resulting unique molecular identifier (UMI) matrices, including three files (barcodes. tsv, features.tsv, and matrix, mtx), were used to generate Seurat objects as previously described in Hafemeister, C. and Satija, R. {Normalization and variance stabilization of single-cell RNA-seq data using regularized negative binomial regression. Genome Biol. 2019 Dec 23;20(1):296) incorporated herein in its entirety. For each sample, cells with >3,000 detected genes, total RNA counts >10,000, fewer than 200 detected genes or >10% mitochondrial RNA (mtRNA) content were filtered and removed. After quality filtering, 16,560 cells from leiomyoma samples and 11,354 cells from their myometrial counterparts were subjected to subsequent Seurat analysis. This included “NormalizeData”, “FindVariableGenes” (to identify highly variable genes), “RunPCA” (to estimate principal components (PCs), and “RunUMAP” (to visualize the distribution of cells across dimensions 1 to 20). Cell types were annotated based on differentially expressed genes (DEGs) and well-established cellular markers from the literature, as shown in Table 1, using Semisupervised Category Identification and Assignment (SCINA). RidgePlots were generated in RStudio and used to display gene expressions, with the average expression values represented by a color scale and the percentage of cells expressing each gene.Table 1 - Cellular MarkersTo infer patterns of intercellular communication, CellChat (RRID:SCR_021946) was used to quantitatively identify signaling pathways based on the average expression values for ligands in putative donor cell and receptor in putative recipient cells, as well as their cofactors. CellChat objects were created with the normalized expression matrix via the create CellChat function. Subsequent processes, including “identify over expressed Genes”, “identify over expressed interaction”, “project data”, “compute communProb”, “filter Communication (min. cells = 10)”, and “compute Commun Prob Pathway” functions, were used to calculate potential ligand-receptor interactions CellChat’s “aggregateNet” function was used to calculate the aggregated cell-cell communication networks as previously described in Jin, S et al. (Inference and analysis of cell-cell communication using CellChat. Nat Commun.2021 Feb 17; 12(1 ): 1088), incorporated herein in its entirety.Statistical AnalysesPaired t-test and ANOVA were used to assess the statistical significance of comparisons made with a threshold for significance set at <0.05. Normality distribution assessed by Shapiro-Wilk test All data are presented as mean ± SEM. Statistical analyses were performed and visualized using GraphPad Prism 10 (GraphPad, Inc, San Diego, CA, USA; RRID:SCR_002798)ConclusionThe inventors demonstrate, for the first time, that COL12A1 as a key, mechanosensitive ECM regulator in leiomyomas with both structural and signaling roles in leiomyomas. It plays a dual role in structural matrix modelling and cell signaling, supports leiomyoma growth and ECM organization The resulting data, demonstrated here, point to a previously unappreciated role for COL12A1 in maintaining the organization and survival of leiomyomas in addition to directly promoting the proliferation of uterine myocytes. The inventors have shown that exposure of myometrial cells to simulated microgravity, a model of mechanical unloading, resulted in significant upregulation of COL12A1 at the promoter level, thus suggesting that COL12A1 increase may be a part of compensatory response to ECM disorganization or biomechanical stress which part of a feed-forward loop that could accelerate leiomyoma growth. Given its consistent overexpression in leiomyomas, mechanosensitive regulation, and impact on proliferation, COL12A1 represents a promising non-hormonal, localized therapeutic target for the treatment of uterine leiomyomas.The disclosures of all publications cited above are expressly incorporated herein by reference, each in its entirety, to the same extent as if each were incorporated by reference individually.It is also to be understood that the following claims are intended to cover all of the generic and specific features of the invention herein described, and all statements of the scope of the invention which, as a matter of language, might be said to fall there between. Now that the invention has been described,

Claims

What is claimed is:

1. A method of treating a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a composition, the composition comprisinga therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding for at least one fibril-associated collagen with interrupted triple helices (FACITs) wherein the at least one gene or gene expression product is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof; anda pharmaceutically acceptable carrier;wherein the therapeutically effective amount of the composition downregulates expression of the at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to treat the uterine leiomyoma.

2. A method of preventing growth of a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a composition, the composition comprisinga therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding for at least one fibril-associated collagen with interrupted triple helices (FACITs) wherein the at least one gene or gene expression product encoding the at least one FACIT is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof; anda pharmaceutically acceptable carrier;wherein the therapeutically effective amount of the composition downregulates expression of the at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to prevent growth of the uterine leiomyoma.

3. A method of inhibiting tumor cell proliferation in a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a composition, the composition comprisinga therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding for at least one fibril-associated collagen with interrupted triple helices (FACITs) wherein the at least one gene or gene expression product encoding the at least one FACIT is selected from collagen type XII alpha 1 chain (COL12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A7), and combinations thereof; anda pharmaceutically acceptable carrier;wherein the therapeutically effective amount of the composition downregulates expression of the at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to inhibit tumor cell proliferation in the uterine leiomyoma.

4. A method of decreasing accumulation of extracellular matrix (ECM) in a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a composition, the composition comprisinga therapeutically effective amount of a therapeutic agent capable of downregulating expression of at least one gene or gene expression product encoding for at least one fibril-associated collagen with interrupted triple helices (FACITs) wherein the at least one gene or gene expression product encoding the at least one FACIT is selected from collagen type XIIalpha 1 chain (C0L12A1), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), and combinations thereof; anda pharmaceutically acceptable carrier;wherein the therapeutically effective amount of the composition downregulates expression of the at least one gene or gene expression product encoding the at least one FACIT in the uterine leiomyoma in the patient to decrease the accumulation of ECM in the uterine leiomyoma.

5. The method of any of claims 1 -4, wherein the therapeutic agent is an RNA interference agent targeting the at least one gene or gene expression product.

6. The method of any of claims 1 -5, wherein the at least one gene or gene expression product is COL12A1.

7. The method of any of claims 1-4 or 6, wherein the therapeutic agent is selected from an RNA interference agent targeting COL12A1, omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, and nilotinib.

8. The method of any of claims 1 -7, wherein the patient is pre-menopausal.

9. A method of treating a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing production / synthesis of and / or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT);wherein the at least one FACIT is selected from the group consisting of collagen type XII, collagen type IX, collagen type XXII, and combinations thereof; wherein the therapeutically effective amount of the therapeutic agent reduces the production of and / or amount of the at least one FACIT in the uterine leiomyoma in the patient to treat the uterine leiomyoma.

10. A method of inhibiting tumor cell proliferation in a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing production / synthesis of and / or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT);wherein the at least one FACIT is selected from the group consisting of collagen type XII, collagen type IX, collagen type XXII, and combinations thereof; wherein the therapeutically effective amount of the therapeutic agent inhibits tumor cell proliferation in the uterine leiomyoma.

11. A method of preventing growth of a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing production / synthesis of and / or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT);wherein the at least one FACIT is selected from the group consisting of collagen type XII, collagen type IX, collagen type XXII, and combinations thereof; wherein the therapeutically effective amount of the therapeutic agent prevents growth of the uterine leiomyoma.

12. A method of decreasing accumulation of extracellular matrix (ECM) in a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing production / synthesis of and / or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT);wherein the at least one FACIT is selected from the group consisting of collagen type XII, collagen type IX, collagen type XXII, and combinations thereof; wherein the therapeutically effective amount of the composition decreases accumulation of extracellular matrix (ECM) in the uterine leiomyoma.

13. A method of reducing structural integrity and / or reducing stiffness in a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing production / synthesis of and / or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT);wherein the at least one FACIT is selected from the group consisting of collagen type XII, collagen type IX, collagen type XXII, and combinations thereof; wherein the therapeutically effective amount of the composition reduces the structural integrity and / or the stiffness of the uterine leiomyoma.

14. A method of preventing fibrosis in a uterine leiomyoma in a patient in need thereof comprising:administering to the patient a therapeutically effective amount of a therapeutic agent capable of reducing production / synthesis of and / or amount of at least one fibril-associated collagen with interrupted triple helices (FACIT);wherein the at least one FACIT is selected from the group consisting of collagen type XII, collagen type IX, collagen type XXII, and combinations thereof; wherein the therapeutically effective amount of the composition prevents fibrosis in the uterine leiomyoma.

15. The method of any of claims 9-14, wherein the at least one FACIT is collagen type XII.

16. The method of any of claims 9-15, wherein the therapeutic agent is an RNA interference agent targeting at least one gene or gene expression product selected from collagen type XII alpha 1 chain (COL12AT), collagen type IX alpha 2 chain (COL9A2), collagen type XXII alpha 1 chain (COL22A1), or combinations thereof.

17. The method of any of claims 9-16, wherein the therapeutic agent is omidenepag isopropyl (OMDI), omidenepag (OMD), caffeine, decitabine, U0126, palbociclib, geldanamycin, rifocin, eupachlorin, nanaomycin, or nilotinib.

18. The method of any of claims 9-17, wherein the patient is pre-menopausal.

9. The method of any of claims 9-18, further comprising a pharmaceutically acceptable carrier wherein the therapeutic agent and the pharmaceutically acceptable carrier form a composition.

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