Compositions comprising EP300 modulators and methods of using the same

Abstract

The disclosure relates to methods of stimulating wound healing comprising administering an effective amount of a modulator of acetyltransferase to the subject. The disclosure also relates to methods of treating or preventing fibrosis, reducing severity of scarring or preventing scarring, inhibiting acetyltransferase, reducing a population of profibrotic fibroblasts, an enhancing SOX10 and / or SOX9 expression in a wound in need thereof comprising administering an EP300 modulator to a subject in need thereof. The disclosure also relates to compositions and pharmaceutical compositions comprising an effective amount of an EP300 modulator.

Description

COMPOSITIONS COMPRISING EP300 MODULATORS AND METHODS OF USING THE SAMECROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 668,162, which was filed July 6. 2024, is titled “Compositions Comprising EP300 Modulators and Methods of Using the Same,” and is incorporated herein by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under grant numbers R01- GM136659, U24DE029463, R01-DE032677, R01-AR081343, RM1-HG007735, awarded by the National Institutes of Health. The government has certain rights in this invention.SEQUENCE LISTING

[0003] The electronic sequences filed herewith, titled “STFD-009-PCT_SL.xml,” created June 23, 2025, and having a file size of 24,411 bytes is incorporated herein by reference in its entirety.FIELD

[0004] The disclosure relates to methods of stimulating wound healing comprising administering an effective amount of a modulator of acetyltransferase to the subject. The disclosure also relates to methods of treating or preventing fibrosis, reducing severity of scarring or preventing scarring, inhibiting acetyltransferase, reducing a population of profibrotic fibroblasts, an enhancing SOX10 and / or SOX9 expression in a wound in need thereof comprising administering an EP300 modulator to a subject in need thereof.BACKGROUND

[0005] Fibrosis is a major cause of morbidity and mortality worldwide, accounting for up to 45% of all deaths in the United States.1In the skin, scarring can result in significant growth restriction, psychosocial sequelae, and functional loss. Despite its widespread prevalence and negative impacts, no current clinical treatments fully prevent or reverse this fibrotic process.

[0006] The ability to heal with a regenerative or fibrotic outcome varies across organisms, developmental stages, and anatomical locations.2'4Elucidating the mechanisms underlyingthese differential wound healing responses can shed light into broadly effective anti-fibrotic therapies. It is well known in clinical practice that facial wounds heal more quickly and with less scaring than cutaneous wounds on the trunk and extremities. Sutures placed to close facial wounds, for example, can be removed earlier than sutures placed on the trunk or extremities,5and there is a higher risk of hypertrophic scarring on the trunk or limbs than on the face.6Various factors can contribute to differential wound healing across anatomical locations, including dermal thickness, mechanical tension, vascular supply, closure technique, and postoperative wound care.3'5’7’9Although these can all impact the degree of scar formation, the molecular mechanisms underlying differential healing outcomes remain unknown.

[0007] Increasing evidence for fibroblast heterogeneity suggests there may be fibroblast- intrinsic properties underlying differential scar formation.10,11Fibroblasts from disparate human anatomical sites contain unique positional identities specified by distinct gene expression patterns, including HOX genes important to axial patterning in embry ogenesis.12,13Furthermore, dermal fibroblasts have different embryonic origins depending on their location: facial, scalp, ventral, and dorsal fibroblasts derive primarily from the cranial neural crest, cephalic mesoderm, lateral plate mesoderm, and paraxial mesoderm, respectively.14These fibroblast origins may in turn influence fibrotic behavior in postnatal life. Fibrogenic lineages of mouse fibroblasts from dorsal and ventral skin, for example, are defined by different transcriptional programs specified by Engrailed- 1 and Paired-related homeobox 1 expression, respectively.10,11,15Thus, it is possible that facial fibroblasts, in turn, derive their regenerative potential from their neural crest origin.

[0008] The neural crest is a multipotent stem cell-like population that emerges from the dorsal neural plate and forms the craniofacial skeleton and connective tissue.16A cascade of molecules directs the migration of neural crest cells from the neural plate, and both neural crest cells and their derivatives show remarkable plasticity.17’21For example, after injury to the jaw bone in the face, skeletal stem cells revert to a more progenitor 'neural crest-like state' to promote regeneration of new bone,22indicating the reparative potential of adult neural-crest progenitors. However, the regenerative potential of neural-crest-derived fibroblasts postnatally has been so far unexplored.SUMMARY OF EMBODIMENTS

[0009] The disclosure relates to a method of stimulating wound healing in a subject in need thereof. In some embodiments, the method comprises administering an effective amount of amodulator of acetyltransferase to the subject. In some embodiments, the EP300 modulator is chosen from: (i) SLIT2 or a functional variant thereof; (ii) EID1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv) an EP300 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the EP300 modulator is an EP300 inhibitor comprising one or a combination of EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS- 9300, or NEO3734, or a pharmaceutically acceptable salt thereof. In some embodiments, the modulator of acetyltransferase is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0010] The disclosure also relates to methods of treating or preventing fibrosis in a subject in need thereof. In some embodiments, the method comprises administering an effective amount of an EP300 modulator to the subject. In some embodiments, the EP300 modulator is chosen from: (i) SLIT2 or a functional variant thereof; (ii) EID1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv) an EP300 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the EP300 modulator is an EP300 inhibitor comprises at least one of EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS-9300, or NEO3734, or a pharmaceutically acceptable salt thereof. In some embodiments, the modulator of acetyltransferase is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof. . In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0011] In some embodiments the disclosure relates to a method of reducing severity' of scarring or preventing scarring in a subject in need thereof, The method comprises administering an effective amount of an EP300 modulator to the subject. . In some embodiments, the EP300 modulator is chosen from: (i) SLIT2 or a functional variant thereof; (ii) EID1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv)an EP300 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the EP300 modulator is an EP300 inhibitor comprising one or a combination of: EP300i. Y08197, CCS 1477, CPI-1612, DS 17701585, DS-9300, orNEO3734, and a pharmaceutically acceptable salt thereof.

[0012] In some embodiments the disclosure relates to a method of inhibiting acetyltransferase in a subject in need thereof. The method comprises administering to the subject an EP300 modulator. In some embodiments, the EP300 modulator is chosen from: (i) SLIT2 or a functional variant thereof; (ii) EID1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv) an EP300 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the EP300 inhibitor comprises at least one of EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS-9300. or NEO3734, or a pharmaceutically acceptable salt thereof. In some embodiments, the EP300 modulator is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0013] In some embodiments the disclosure relates to a method of differentiating a fibroblast comprising contacting the fibroblast with an effective amount of an EP300 modulator. In some embodiments, the EP300 modulator is chosen from: (i) SLIT2 or a functional variant thereof; (ii) EID1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv) an EP300 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the EP300 modulator is an EP300 inhibitor comprising one or a combination of: EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS-9300, orNEO3734, and a pharmaceutically acceptable salt thereof. In some embodiments, the method is performed in vitro. In some embodiments, the method is performed in vivo within a mammal and further comprises administering to the mammal an effective amount of the EP300 modulator.

[0014] The disclosure relates to a method of reducing a population of profibrotic fibroblasts in a subject in need thereof. The method comprises administering to the subject an EP300 modulator. In some embodiments, the EP300 modulator is chosen from: (i) SLIT2 or a functional variant thereof; (ii) EID 1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv) an EP300 inhibitor or a pharmaceutically acceptable salt thereof. Insome embodiments, the EP200 modulator is an EP300 inhibitor chosen from one or a combination of: EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS-9300, orNEO3734, and a pharmaceutically acceptable salts thereof. In some embodiments, the EP300 modulator is administered by oral adminstrati on, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0015] The disclosure also relates to methods of enhancing SOXIO and / or SOX9 expression in a wound of a subject in need thereof. The method comprises administering to the wound of the subject an EP300 modulator. In some embodiments, the EP300 modulator is chosen from: (i) SLIT2 or a functional variant thereof; (ii) EID1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv) an EP300 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the EP200 modulator is an EP300 inhibitor comprising one or a combination of: EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS- 9300, orNEO3734, and a pharmaceutically acceptable salts thereof. In some embodiments, the EP300 modulator is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0016] In some embodiments the disclosure relates to a pharmaceutical composition comprising: (i) an effective amount of an EP300 modulator or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier. In some embodiments, the effective amount is from about 10 grams to about 1000 grams of the EP300 modulator. In some embodiments, the EP300 modulator is chosen from one or a combination of: (i) SLIT2 or a functional variant thereof; (ii) EID1 or a functional variant thereof; (iii) ROBO2 or a functional variant thereof; and (iv) an EP300 inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from: EP300i. Y08197, CCS1477, CPI-1612, DS17701585, DS-9300, NEO3734, and pharmaceutically acceptablesalts thereof. In some embodiments, the EP300 modulator is an agent that increases expression of SLIT2, ROBO2 and / or EID1.

[0017] In some embodiments, the disclosure relates to an isolated fibroblast comprising or cell line comprising a fibroblast that comprises Robo2+ or a functional variant thereof. In some embodiments, the fibroblast or cell line further comprises one or a combination of: Cdh4+, Ncaml+, and Gpc3 or a functional fragment thereof. In some embodiments, the fibroblast comprises SOX9 and / or SOXIO. In some embodiments, the fibroblast or cell line is frozen at about -20 degrees Celsius or about -212 degrees Celsius.

[0018] The disclosure also relates to pharmaceutical compositions comprising an effective amount of an isolated fibroblast comprising or cell line comprising a fibroblast that comprises Robo2 or a functional variant thereof and a pharmaceutically acceptable carrier. In some embodiments, the fibroblast or cell line further comprises one or a combination of: Cdh4+. Ncaml+, and Gpc3 or a functional fragment thereof. In some embodiments, the fibroblast comprises SOX9 and / or SOXIO. In some embodiments, the fibroblast or cell line is frozen at about -20 degrees Celsius or about -212 degrees Celsius.

[0019] In some embodiments the disclosure relates to a method of preventing or treating fibrosis in a subject in need thereof. In some embodiments, the method comprises administering to the subject an effective amount of a pharmaceutical composition comprising an effective amount of an isolated fibroblast comprising or cell line comprising a fibroblast that comprises R bo2 or a functional variant thereof and a pharmaceutically acceptable carrier. In some embodiments, the fibroblast or cell line further comprises one or a combination of: Cdh4+, Ncaml+. and Gpc3 or a functional fragment thereof. In some embodiments, the fibroblast comprises SOX9 and / or SOXIO. In some embodiments, the fibroblast or cell line is frozen at about -20 degrees Celsius or about -212 degrees Celsius. In some embodiments, the method comprises administering to the subject an effective amount of an isolated fibroblast comprising or cell line comprising a fibroblast that comprises Robo2+ or a functional variant thereof. In some embodiments, the fibroblast or cell line further comprises one or a combination of: Cdh4+. Ncaml+, and Gpc3 or a functional fragment thereof. In some embodiments, the fibroblast comprises SOX9 and / or SOXIO. In some embodiments, the fibroblast or cell line is frozen at about -20 degrees Celsius or about -212 degrees Celsius. In some embodiments, the pharmaceutical composition or the fibroblasts is administered by oral adminstrati on, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration,intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0020] The disclosure relates to a method of promoting wound healing in a subj ect in need thereof. In some embodiments, the method comprises administering to the subject an effective amount of a pharmaceutical composition comprising an effective amount of an isolated fibroblast comprising or cell line comprising a fibroblast that comprises Robo2+ or a functional variant thereof and a pharmaceutically acceptable carrier. In some embodiments, the fibroblast or cell line further comprises one or a combination of: Cdh4+. Ncaml+, and Gpc3 or a functional fragment thereof. In some embodiments, the fibroblast comprises SOX9 and / or SOX 10. In some embodiments, the fibroblast or cell line is frozen at about -20 degrees Celsius or about -212 degrees Celsius. In some embodiments, the method comprises administering to the subject an effective amount of an isolated fibroblast comprising or cell line comprising a fibroblast that comprises Robo2+ or a functional variant thereof. In some embodiments, the fibroblast or cell line further comprises one or a combination of: Cdh4+, Ncam.l+. and Gpc3 or a functional fragment thereof. In some embodiments, the fibroblast comprises SOX9 and / or SOX10. In some embodiments, the fibroblast or cell line is frozen at about -20 degrees Celsius or about -212 degrees Celsius. In some embodiments, the pharmaceutical composition or the fibroblast is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the pharmaceutical composition is administered directly into the wound of the subject. In some embodiments, the method comprises administering from about 10,000 to about 10.000.000 cells.BRIEF DESCRIPTION OF DRAWINGS

[0021] FIG 1 A illustrates a schematic of the SLIT2-ROBO2-EID1-EP300 signaling axis in the context of dermal fibrosis.

[0022] FIGS. 2A-2C illustrate a four-site murine dermal excisional wounding model. FIG. 2A illustrates a schematic of the "Tour-site murine wounding model” to evaluate healing outcomes involving fibroblasts from four different embryonic origins. FIG. 2B illustrates a hematoxylin and eosin (H&E) and Masson’s Trichrome staining of unwounded skin and skin at POD 14 on the face, scalp, ventrum, and dorsum, with quantification of dermal thickness (by H&E) and degree of collagen deposition (by Trichrome) (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001). Scars are contoured by dotted lines. Vertical black dotted lines on H&E denote dermal thickness. FIG. 2C illustrates IHC staining for vimentin (VIM), alpha smooth muscle actin (aSMA), collagen 1 (COL1), and collagen 3 (COL3) of unwounded and POD 14 skin on the face, scalp, ventrum, and dorsum. Bar graphs show quantification of staining intensity for ?SMA, COL1, and COL3 (***p < 0.001. ****p < 0.0001).

[0023] FIG. 3 A illustrates a schematic of transplantation of skin grafts from the face, scalp, ventrum, and dorsum of RFP donor mice into the dorsum of wildtype (B6) recipient mice.

[0024] FIGS. 3B and 3C illustrate IHC staining and quantification of co-localization of COL1 with RFP (donor cells) in the skin from the four sites engrafted onto the dorsum of recipient mice 14 days after (POD 14) incisional wounding (*p < 0.05. **p < 0.01, ***p < 0.001).

[0025] FIG. 4A illustrates a schematic of isolation and transplantation of fibroblasts from the face, scalp, ventrum, and dorsum of GFP donor mice into the dorsum of wildtype (B6) recipient mice.

[0026] FIGS. 4B and 4C illustrate IHC staining and quantification of co-localization of COL1 with GFP (injected fibroblasts) in the dorsal dermis of recipient mice 10 days after (POD 10) injection with fibroblasts from the four sites of donor mice (*p < 0.05, **p < 0.01, ***p < 0.001).

[0027] FIGS. 5A-5G illustrate an excisional wound model and scRNA-seq and analyses of fibroblasts from four-site murine dermal. 5A illustrates a schematic of fibroblast isolation and scRNA-seq workflow. FIG. 5B illustrates UMAP of fibroblast clusters from unwounded (UW) skin, and at POD 7 and POD 14 after wounding across four sites; transcriptionally distinct clusters at POD 14 are labeled and used for subsequent analyses. FIG. 5C illustrates distribution of fibroblasts from different sites across scRNA-seq clusters on the left, and CytoTRACE analysis of cellular differentiation states of fibroblasts at POD 14, with clusters numbered, on the right. FIG. 5D illustrates Gene Ontology pathway analysis clusters 0, 2, and 4 revealed processes implicated in neural crest biology and regeneration. FIG. 5E illustrates highly expressed genes in facial fibroblasts and clusters 0, 2, 4 encoding cell surface proteinsinvolved neural crest cell signaling. FIG. 5F illustrates pseudotime analysis of Cdh4, Gpc3, and Ncaml gene expression patterns. Robo2 expression was greatest in cluster 2. FIG. 5G illustrates FACS of fibroblasts from different wound sites at POD 14 sorted by their cell surface protein levels. Percentage of fibroblasts with R0B02 was the highest in facial wounds compared to dorsal, ventral, and scalp wounds (*p<0.05).

[0028] FIGS. 6A-6I illustrate healing with reduced fibrosis and that Robo2 in fibroblasts is necessary and sufficient to promote wound. FIG. 6A illustrates a schematic of FACS isolation of ROBO2-high (ROBO2+) and ROBO2-low (ROBO2-) fibroblasts from GFP mice and intradermal injection of ROBO2+ or ROBO2- fibroblasts into dorsal wounds of wildtype mice at the time of wounding. Untreated facial wounds were used as control. Wounds were harvested 14-days after wounding and injection for downstream analyses. FIG. 6B illustrates H&E and Masson's Tri chrome staining of untreated facial wounds (FW) and dorsal wounds injected with ROBO2- (DW ROBO2-) or ROBO2+ (DW ROBO2+) fibroblasts at POD 14, with quantification of dermal thickness (by H&E) and degree of collagen deposition (by Trichrome) (p <0.05). FIG. 6C Top: Picrosirius red staining of collagen fibers of POD 14 wounds; Bottom: UMAP of quantified collagen fiber features for each experimental group. FIG. 6D illustrates IHC staining and quantification of staining intensity (bar graph) of ROBO2, COL1, SOXIO of POD 14 wounds (*p< 0.05); Nuclear staining (DAPI) and GFP+ ROBO2- or ROBO2+ fibroblasts. FIG. 6E illustrates a schematic of CRISPR-mediated Robo2 knockout or Robo2 overexpression or control CRISPR (with scrambled gRNA) in cultured mouse facial fibroblasts and injection of these fibroblasts into dorsal wounds. Untreated facial and dorsal wounds were used as additional comparison groups. All wounds were harvested at POD 14 for subsequent analyses. FIG. 6F illustrates H&E and Masson’s Tri chrome staining of POD 14 untreated facial wounds (FW), untreated dorsal wounds (DW), dorsal wounds injected with CRISPR-control fibroblasts (DW Robo2 Scr [scrambled]), dorsal wounds injected with Robo2 -knockout fibroblasts (DW Robo2 KO), and dorsal wounds injected with Robo2- overexpression fibroblasts, with quantification of dermal thickness (by H&E) and degree of collagen deposition (by Tri chrome) (*p <0.05). G) Left: Picrosirius red staining of collagen fibers of POD 14 wounds. Right: UMAP of quantified collagen fiber features for each experimental group. H) IHC staining and quantification of staining intensity (bar graph) of SOX 10 and COL 1 in POD 14 wounds (*p< 0.05); ROBO2 IHC staining and nuclear staining (DAPI). I) Diagram of ROBO2’s role in dermal fibrosis.

[0029] FIGS. 7A-7E illustrate quantification of fibrotic features of wounds with ROBO2+or ROBO2" fibroblasts. FIG. 7A illustrates fluorescence-activated cell sorting (FACS) offibroblasts by lineage-negative gating (left), then sorting of ROBO2 fibroblasts (arrowhead) and ROBO2 fibroblasts (right) from facial skin. FIG. 7B illustrates a bar graph showing RT- qPCR quantification of collagen I (Coll), collagen III (Col3), and a-smooth muscle actin (aSMA) (**p < 0.01) and FIG. 7C illustrates ELISA of collagen levels from fibroblasts of POD 14 untreated facial wounds (FW), dorsal wounds injected with ROBO2" fibroblasts (DW ROBO2 ), and dorsal wounds injected with ROBO2+fibroblasts (DW ROBO2+) (*p < 0.05, **p < 0.01). FIG. 7D illustrates a bar graph showing RT-qPCR quantification of collagen I (Coll), collagen III (Col3), and a-smooth muscle actin (aSMA) (*p < 0.05) and FIG. 7E illustrates ELISA of collagen levels in wound supernatant of POD 14 untreated facial wounds (FW), untreated dorsal wounds (DW), dorsal wounds inj ected with Robo2 -knockout fibroblasts (DW Robo2 KO), dorsal wounds injected with control fibroblasts (DW Robo2 Scr), and dorsal wounds injected with Robo2-overexpressing fibroblasts (DW Robo2 OE) (*p < 0.05).

[0030] FIGS. 8A-8D illustrate that SLIT2 suppresses the fibrotic activity’ of dorsal fibroblasts in vitro. FIG. 8A illustrates an immunohistochemical staining (left) and quantification (right) of staining intensity of ROBO2, SOX 10, and aSMA in cultured facial fibroblasts or dorsal fibroblasts receiving SLIT2 treatment (SLIT2+) or vehicle control (SLIT2- ). Nuclear staining (DAPI). FIG. 8B illustrates FACS isolation of cultured facial fibroblasts and dorsal fibroblasts receiving SLIT2 treatment (SLIT2+) or vehicle control (SLIT2-) by SOX10 levels (*p < 0.05). FIG. 8C illustrates a bar graph showing RT-qPCR quantification of expression levels of collagen I (Coll), collagen III (Col3), and a-smooth muscle actin (a-SMA) in cultured facial fibroblasts and dorsal fibroblasts receiving SLIT2 treatment (SLIT2+) or vehicle control (SLIT2-) (*p < 0.05). FIG. 8D illustrates ELISA quantification of collagen 1 and TGF-betal levels in the conditioned media of cultured facial fibroblasts and dorsal fibroblasts receiving SLIT2 treatment (SLIT2+) or vehicle control (SLIT2-).

[0031] FIGS. 9A-9G illustrate that SLIT2 treatment reduces scarring in dorsal wounds. FIG. 9A illustrates a schematic of treating dorsal wounds with SLIT2 or vehicle control at the time of wounding and wound harvesting at POD 14 for downstream analyses. FIG. 9B illustrates H&E and Masson’s Trichrome staining of POD 14 untreated facial wounds (FW), dorsal wounds treated with vehicle control (DW SLIT2-), and dorsal wounds treated with SLIT2 (DW SLIT2+), with quantification of dermal thickness (by H&E) and degree of collagen deposition (by Trichrome) (*p < 0.05). FIG. 9C illustrates Top: Picrosirius red staining of collagen fibers of healed wounds at POD 14; and bottom: UMAP of quantified collagen fiber features for each experimental group. FIG. 9D illustrates an IHC staining and quantification of staining intensity (bar graph) of ROBO2, COLL and SOX10 in healed wounds at POD 14 (*p< 0.05); nuclear staining (DAPI). FIG. 9E illustrates expression levels of fibrosis markers (Coll, Col3, aSMA) in healed wounds at POD 14 (*p < 0.05). FIG. 9A illustrates ELISA quantification of collagen 1 levels in the wound supernatant of experimental groups (***p < 0.001). FIG. 9G illustrates a diagram of SLIT2’s role in dermal fibrosis.

[0032] FIGS. 10A and 10B illustrate that Eidl is enriched in facial wounds. FIG. 10A illustrates violin plots of Eidl, Trspl, Tcf712, and Glil expression by Seurat fibroblast cluster (top) and anatomical location (bottom) of fibroblasts derived from wounds at post-operative day 14 (POD 14). FIG. 10B illustrates IHC staining and quantification of co-localization of ROBO2 and EID1 in healed facial and dorsal wounds at POD 14; nuclear staining (DAPI) (***p < 0.001).

[0033] FIGS. 11A-11C illustrate an epistatic relationship of Robol and Eidl. FIG. HA illustrates a schematic of primary culture of facial fibroblasts, CRISPR-mediated Robol or Eidl knockout, and downstream analyses. FIG. 11B illustrates IHC staining and quantification of staining levels of ROBO2, EID1, EP300, and COL1 in control, Robol knockout (Robol KO), and E / l knockout (Eidl KO) cultured facial fibroblasts (*p < 0.05). FIG. 11C illustrates fluorescence-activated cell sorting (FACS) of control. Robol KO. and Eidl KO cultured facial fibroblasts by ROBO2, EID1, and EP300 levels, showing percentage of fibroblasts positive for each marker (*p < 0.05).

[0034] FIGS. 12A-12E illustrate Eidl expression in fibroblasts is sufficient to promote wound healing with reduced fibrosis. FIG. 12A illustrates a schematic of CRISPR-mediated Eid\ knockout or Eidl overexpression or control CRISPR (with scrambled gRNA) in cultured mouse facial fibroblasts and injection of these fibroblasts into dorsal w ounds. Untreated facial and dorsal wounds were used as additional comparison groups. All healed wounds were harvested at POD 14 for subsequent analyses. FIG. 12B illustrates H&E and Masson's Trichrome staining of POD 14 untreated facial wounds (FW), untreated dorsal wounds (DW), dorsal wounds injected with CRISPR-control fibroblasts (DW Eidl Scr [scrambled]), dorsal wounds injected with Eidl -knockout fibroblasts (DW Eidl KO), and dorsal wounds injected with Eidl -overexpression fibroblasts (DW Eidl OE), with quantification of dermal thickness (by H&E) and degree of collagen deposition (by Tri chrome) (ns = not significant). FIG. 12D illustrates: Left: Picrosirius red staining of collagen fibers of healed wounds at POD 14; and Right: UMAP of quantified collagen fiber features for each experimental group. FIG. 12D illustrates IHC staining and quantification of staining intensity (bar graph).

[0035] FIGS. 13A-13D illustrate CRISPR-mediated Eidl knockout and overexpression in fibroblasts and injection into dorsal wounds. FIG. 13A illustrates IHC staining andquantification of EID1, COL1 levels in facial fibroblasts, dorsal fibroblasts, and CRISPR- mediated Eidl -knockout (Eidl KO) dorsal fibroblasts at 48 hours (*p < 0.05). FIG. 13B illustrates fluorescence-activated cell sorting (FACS) analysis of facial fibroblasts, dorsal fibroblasts, Eidl KO fibroblasts by SOXIO levels. FIG. 13C illustrates a bar graph showing RT-qPCR quantification of expression levels of collagen I, collagen III, and aSMA and FIG. 13D illustrates ELISA of collagen levels in wound supernatant of POD 14 untreated facial wounds (FW), untreated dorsal wounds (DW). dorsal wounds injected with CRISPR-mediated Eid\ - knockout fibroblasts (DW Eidl KO), dorsal wounds injected with CRISPR control fibroblasts (DW Eidl Scr [scrambled]), and dorsal wounds injected with CRISPR- mediated Ei dl -overexpressing fibroblasts at the time of wounding.

[0036] FIGS. 14A-14C illustrate lentivirus-mediated Eidl overexpression at early stages of wound healing reduces scarring in dorsal wounds. FIG. 14A illustrates representative H&E images of dorsal wounds at POD 30 (Colal-CreERT; Ai9 mice) injected with Eidl overexpression control lentivirus (DW Eztil-OE control), Eidl overexpression lentivirus (DW Eidl-OE), Eidl -knockdown control lentivirus (DW Eidl -KD control), and Eidl - knockdown lentivirus (DW Ezdl-KD) 3 days prior and 2 days after wounding (POD- 3 / +2); scars bounded by dotted lines. FIG. 14B illustrates representative Picrosirius red images ofEz l-OE control, DW Eidl -OE, DW Eid 1 -KD control and DW Ei dl -KD with UMAP quantification (bottom) at POD -3 / +2. FIG. 14C illustrates IHC staining of SOXIO in healed wounds at POD 14 for each experimental group; nuclear staining (DAPI).

[0037] FIGS. 15A-15C lentivirus-mediated Eidl overexpression at late stages of wound healing reduces scarring in dorsal wounds. FIG. 15 A illustrates representative H&E images of dorsal wounds at POD 30 (Colal-CreERT; Ai9 mice) injected with Eidl overexpression control lentivirus (DW EzJl-OE control), Eidl overexpression lentivirus (DW Ez l-OE), Eidl- knockdown control lentivirus (DW Ezdl-KD control), and Eidl- knockdown lentivirus (DW Ez l-KD) 7 and 10 days after wounding (POD 7 / 10); scars bounded by dotted lines. FIG. 15B illustrates representative Picrosirius red images of Eidl-OE control, DW Ez l-OE, DW Eidl- KD control and DW A’zc / I -KD with UMAP quantification (bottom) at POD 7 / 10. FIG. 15C illustrates IHC staining of SOXIO in healed wounds at POD 14 for each experimental group; nuclear staining (DAPI).

[0038] FIGS. 16A-16I illustrate in vitro and in vivo small-molecule inhibition of EP300 attenuates scarring phenotype. FIG. 16A illustrates a diagram of in vitro fibroblast experiments with EP300i treatment and downstream analyses FIG. 16B illustrates Left: IHC staining of cultured control facial fibroblasts, control dorsal fibroblasts, and dorsal fibroblasts 48 hoursafter EP300 inhibitor treatment, showing EID1, Collagen type I and nuclear staining (DAPI); and Right: Percent coverage of EP300+ cells within imaging field of cultured facial fibroblasts, culture dorsal fibroblasts, and cultured dorsal fibroblasts treated with EP300i (*p < 0.05). FIG. 16C illustrates RT-qPCR quantification of Coll expression in cultured facial fibroblasts, dorsal fibroblasts, and EP300i-treated dorsal fibroblasts (*p < 0.05). FIG. 16D illustrates ELISA quantification of COL 1 levels in the media of cultured facial fibroblasts, dorsal fibroblasts, and EP300i -treated dorsal fibroblasts (*p < 0.05). FIG. 16E illustrates fluorescence-activated cell sorting (FACS) of cultured facial fibroblasts, dorsal fibroblasts, and EP300i-treated dorsal fibroblasts by SOX10 levels, showing percentage of fibroblasts positive for each marker (*p < 0.05). FIG. 16F illustrates Left: Picrosirius red staining of collagen fibers of POD 14 wounds; and Right: UMAP of quantified collagen fiber features for each experimental group. FIG. 16G illustrates IHC staining of the POD 14 wounds with SOX9, SOX10, COLL and nuclear staining (DAPI) (left) with quantification (right) (p< 0.05). FIG. 16H illustrates a bar graph showing RT-qPCR quantification of expression levels of collagen I (Coll), collagen III (Col3), and alpha smooth muscle actin (aSMA) (*p < 0.05) and FIG. 161 illustrates ELISA of collagen I levels in the wound supernatant of POD 14 untreated facial wounds (FW), PBS-treated dorsal wounds (DW PBS), and EP300 inhibitor-treated dorsal wounds (DW EP300i) (**p < 0.01).

[0039] FIGS. 17A-17G illustrate EP300 inhibition is sufficient to promote wound healing with reduced scarring. FIG. 17A illustrates a schematic of experiment including three groups - facial wounds (FW). dorsal wounds treated with PBS (DW PBS), and dorsal wounds treated with EP300 inhibitor (EP300i). FIG. 17B illustrates Top: H&E and Masson’s Trichrome staining of untreated facial w ounds (FW) and dorsal w ounds treated dorsal wounds treated with PBS (DW PBS) and dorsal w ounds treated with EP300 inhibitor (EP300i); lines show7the scar area; and Bottom: Quantification of dermal thickness (by H&E) and degree of collagen deposition (by trichrome) (*p <0.05). FIG. 17C illustrates a diagram of EP300 inhibition’s effect on dermal fibrosis. FIG. 17D illustrates a schematic of scRNA-seq experiment showing groups including dorsal unwounded skin (DUW), dorsal wounds treated with PBS (DW PBS) or with EP300 inhibitor (DW EP300i), facial unwounded skin (FUW), and facial wounds treated with PBS (FW PBS) or with EP300 inhibitor (FW EP300i) harvested at post-operative day 14 (POD 14) for single-cell RNA sequencing (scRNA-seq). FIG. 17D illustrates Uniform Manifold Approximation and Projection (UMAP) plots of fibroblast clusters from each experimental sample. FIG. 17F illustrates Pie charts showing proportions of scRNA-seq fibroblast clusters in each experimental sample. FIG. 17G illustrates Gene Ontology pathway analysis of scRNA- seq fibroblast clusters 1-4, and 6.

[0040] FIGS. 18A and 18B illustrate that EP300 inhibition promotes the expression of neural crest cell markers Sox9 and SoxlO in dorsal wounds. FIG. 18A illustrates: Top left: schematic showing PBS (DW PBS) or EP300i treatment (DW EP300i) of dorsal wounds at the time of wounding and harvesting of healed wounds at POD 14 for analysis. Untreated facial wounds were used as an additional comparison group. All wounds made on Sox9-CreERT2; R26mTmG transgenic mice allowing for lineage tracing of Sox9 expression (+GFP) upon tamoxifen treatment. Bottom left: H&E staining of healed wounds of different experimental groups at POD 14, with scars delineated by dotted lines. Right: IHC of GFP levels (Sox9 expression) and EID1 levels in healed wounds at POD 14 of the different experimental groups; bottom panels are higher magnification views of the regions denoted by yellow-dotted insets. FIG. 18B illustrates the same as FIG. 18A, but for SoxlO.

[0041] FIGS. 19A-19D illustrate that in vivo EP300 knockout promotes facial-like dorsal wound healing with reduced fibrosis. FIG. 19A illustrates a schematic showing comparison groups consisting of control transgenic mice (Coll-CreERT2; R26mTmG) with no wounding (DUW), dorsal wounding plus PBS treatment (DW PBS), and dorsal wounding plus EP300 inhibitor treatment (DW EP300i) and heterozygous EP300 knockout transgenic mice (Coll- CreERT2; R26mTmG; Ep300+ / -) with dorsal wounding (DW Ep300 KO). All wounds were harvested at POD 14 for downstream analyses. FIG. 19B illustrates H&E staining of healed wounds at POD 14 of the different experimental groups, with scars delineated by dotted lines. FIG. 19C illustrates UMAP of quantified collagen fiber features based on Picrosirius red collagen staining for each experimental group at POD 14. FIG. 19D illustrates IHC of GFP levels and staining of SOX 10 in healed w ounds at POD 14 for each experimental group; nuclear staining (DAPI).

[0042] FIGS. 20A-20D illustrate single-cell RNA sequencing of facial and dorsal wounds after EP300 inhibition. FIG. 20A illustrates a uniform manifold approximation and projection (UMAP) plot of fine Seurat clustering of all sequenced cells from the scRNA-seq experiment described in FIG. 17D. FIG. 20B illustrates UMAP of all sequenced cells by cell type from the scRNA-seq experiment described in FIG. 17D. FIG. 20C illustrates UMAP of all sequenced cells by cell type in each experimental group from scRNA-seq in FIG. 17D. FIG. 20D illustrates Left: Schematic demonstrating anchor label transfer analysis of FACS-isolated fibroblasts onto fibroblast clusters from the EP300 experiment; and Right: Feature plots of FACS-isolated fibroblasts (clusters 0-5) mapped onto fibroblasts from the EP300 experiment (clusters 1-9).

[0043] FIGS. 21A-21D illustrate a single cell assay for transposase-accessible chromatin of facial and dorsal wounds after EP300 inhibition. FIG. 21 A illustrates a schematic of an experiment showing dorsal unwounded (DUW), dorsal wounds treated with PBS (DW PBS), dorsal treated wounds with EP300 inhibitor (DW EP300i), unwounded facial skin (FUW), facial wounds treated with PBS (FW PBS) and facial wounds treated with EP300 inhibitor (FW EP300i) at post-operative day 14 (POD 14) for single-cell assay for transposase-accessible chromatin (scATAC-seq) analysis. FIG. 21B illustrates a uniform manifold approximation and projection (UMAP) plot of fine Seurat clustering of all sequenced cells colored by Seurat group. FIG. 21 C illustrates UMAP of all sequenced cells colored by cell type. FIG. 21D illustrates UMAP of all sequenced cells by cell type by group.

[0044] FIGS. 22A-22D illustrate fibroblast subpopulations andchromatin accessibility changes bysingle cell assay for transposase-accessible chromatin sequencing in facial and dorsal wounds after EP300 inhibition. FIG. 22A illustrates uniform manifold approximation and projection (UMAP) of fibroblast clusters in all sequenced fibroblasts by single cell sequencing assay for transposase-accessible chromatin (scATAC-seq). FIG. 22B illustrates UMAP of fibroblasts by treatment group. FIG. 22C illustrates Pie charts showing proportions of each fibroblast cluster in different experimental groups. FIG. 22D illustrates Gene Ontology pathway analysis of fibroblast clusters 0, 4, 3, and 5.

[0045] FIGS. 23A-23G illustrate spatial analysis of dorsal and facial wounds by Visium and CODEX. FIG. 23A illustrates a schematic of the Visium and CODEX experiments with unwounded facial skin (FUW), PBS-treated POD 14 facial wounds (FW PBS), EP300i-treated POD 14 facial wounds (FW EP300i), unwounded dorsal skin (DUW), PBS-treated POD 14 dorsal wounds (DW PBS), and EP300i-treated POD 14 dorsal wounds (DW EP300i). FIG. 23B illustrates a schematic of the anchor-based integration imputing scRNA-seq-defined fibroblast identities onto the Visium spatial transcriptomic data. FIG. 23C illustrates spatial plots of the scRNA-seq fibroblast clusters on facial (top) and dorsal (bottom) skin and wounds. FIG. 23D illustrates differential interaction maps in PBS-treated dorsal wounds (DW PBS) vs. PBS- treated facial wounds (FW PBS) (left) and EP300i-treated dorsal wounds (DW EP300i) vs. DW PBS (right). FIG. 23E illustrates Uniform Manifold Approximation and Projection (UMAP) plot of CODEX-defined cell clusters. FIG. 23F illustrates bar graphs quantifying CODEX- defined fibroblast subtype proportions in dorsal and facial wounds (*p < 0.05). FIG. 23G illustrates Histograms of aggregate SLIT2, ROBO2, and EID1 protein levels in CODEX- defined fibroblast clusters (n= 3. unless otherwise stated).

[0046] FIGS. 24A and 24B illustrate CODEX analysis of facial and dorsal wounds. FIG. 24A illustrates representative images of the distribution of CODEX-annotated cell populations on each experimental group. FIG. 24B illustrates bar graphs quantifying fibroblast 3 and CD8- T-cell and fibroblast 4 and epithelial cell interactions (*p < 0.05).

[0047] FIGS. 25A and 25B illustrate that CODEX analysis of facial and dorsal wounds reveals similarities between gene and protein analysis. FIG. 25 A illustrates a ridge (histogram) plot of combined Robo2, Eidl and Slit2 expression levels in scRNA-seq-defined fibroblast clusters. FIG. 25B illustrates uniform manifold approximation and projection (UMAP) plots of ROBO2 and EID 1 -enriched cells by CODEX highlighting fibroblast cluster 4 (top) and of Robo2 and Eid\ -enriched cells by scRNA-seq highlighting fibroblast clusters 5-7 (bottom).

[0048] FIGS. 26A-26G illustrate H3K27ac in mouse dorsal and facial fibroblasts. FIG. 26A illustrates: Top: Schematic of dorsal and facial cultured fibroblasts treated with EP300 inhibitor (+I-CBP112) or vehicle control (-I-CBP112) used for Cut&Tag H3K27ac profiling, with the expected changes in H3K27ac and fibrotic phenotype below (more “+” indicates greater magnitude); and Bottom: Heat maps of H3K27ac levels over all Cut&Tag H3K27ac peaks (rows) in untreated and I-CBP112-treated fibroblasts. FIG. 26B illustrates track plots of H3K27ac levels (y-axis) across 5’ end regions (x-axis, 10Kb range) of selected fibroblast genes in untreated and I-CBP112-treated fibroblasts; transcriptional units denoted by black lines above each plot with arrow pointing to transcription direction; exons denoted by black boxes / hash marks. FIG. 26C illustrates: Left: Volcano plot of differential H3K27ac levels across H3K27ac Cut&Tag peaks in dorsal compared to facial untreated fibroblasts, with peaks associated with selected fibroblast genes color-coded and labelled Eidl, Ep300, and Robo2 as shown; and Right: Bar graph showing numbers of H3K27ac peaks enriched in dorsal or facial untreated fibroblasts, color-coded by their associated genes. FIG. 26D illustrates: Top: Schematic of CRISPR-mediated control or Eidl knockout of dorsal and facial cultured fibroblasts used for Cut&RUN H3K27ac profiling, with the expected changes in H3K27ac and fibrotic phenotype below (more “+” indicates greater magnitude); and Bottom: Heat maps of H3K27ac levels over all Cut&RUN H3K27ac peaks (rows) in control and Eidl- knockdown fibroblasts. FIG. 26E illustrates track plots of H3K27ac levels (y- axis) across 5’ end regions (x-axis, 10Kb range) of selected fibroblast genes in control and Eidl -knockdown fibroblasts. FIG. 26F illustrates avolcano plot of differential H3K27ac levels across H3K27ac Cut&RUN peaks in Eidl -knockdown compared to control facial fibroblasts, with peaks associated with selected fibroblast genes coded as shown. FIG. 26G illustrates a diagram showing the relationship between H3K27ac-mediated chromatin states and skin scarring.

[0049] FIGS. 27A-27G illustrate FIGS. 27A) H&E and Masson's Trichrome staining of human unwounded facial and back skin and human facial and back scars (scale bar, 200pm); n = 3 biological replicates per group. FIGS. 27B) IHC staining for ROBO2 and EID1 in human facial and back unwounded skin (FUW, DUW) and facial and back scars (FS, DS) (scale bar, 20pm), with quantification of proportion of cells (DAPI+) with ROBO2; EID1 co-localization (**p < 0.01); n = 3 biological replicates per group. FIGS. 27C) Uniform manifold approximation and projection (UMAP) showing fibroblast clusters of integrated scRNA-seq of human fetal60 and adult fibroblasts61,62from previously published data sets. FIGS. 27D) UMAP of the integrated human scRNA-seq data color-coded by fetal or adult origin. FIGS. 27E) Distnbution of fetal or adult cells in each fibroblast cluster in C). FIGS. 27F) Feature and FIGS. 27G) violin plots of SLIT2 and ROBO2 expression levels in the integrated data of previously published human fetal and adult fibroblast scRNA-seq (SLIT2 and ROBO2 exhibited statistically higher expression in fetal compared to adult fibroblasts, adjusted p-value < 0.05).

[0050] FIGS. 28A-28D illustrate quantification of fibrotic features of wounds with ROBO2+ or ROBO2- fibroblasts. FIGS. 28 A) Low-magnification images of picrosirius red staining of postoperative day- 14 (POD- 14) untreated facial wounds (FW), dorsal wounds injected with ROBO2- fibroblasts (DW ROBO2-), and dorsal wounds injected with ROBO2+ fibroblasts (DW ROBO2+); wounded areas bounded by dotted lines (DW ROBO2- scale bar 100pm; all other scale bars, 15pm). FIGS. 28B) Single immunohistochemical staining of ROBO2 (top) or SOX10 (white, bottom) for the experimental groups; DAPI (scale bar, 25pm). FIGS. 28C) Bar graph showing RT-qPCR quantification of collagen I (Coll), collagen III (Col3), and a-smooth muscle actin (aSMA) and FIGS. 28D) ELISA of collagen levels from POD-14 FW, DW ROBO2-, and DW ROBO2+. Between-group statistical differences were assessed using one-way ANOVA with post-hoc Tukey HSD test (ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0051] FIGS. 29A and 29B illustrate quantification of fibrotic features of wounds treated with Robo2 gene-edited fibroblasts. FIG. 29 A) Bar graph showing RT-qPCR quantification of collagen I (Coll), collagen III (Col3), and a-smooth muscle actin (aSMA) and FIG. 29B) ELISA of collagen levels in wound lysate of POD- 14 FW, DW, DW KO Ctrl. DW Robo2 KO, DW OE Ctrl, and DW Robo2 OE. Between-group statistical differences were assessed using one-way ANOVA with post- hoc Tukey HSD test (ns, not significant, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0052] FIGS. 30A and 30B illustrate that SLIT2 suppresses the fibrotic activity of dorsal fibroblasts in vitro. FIG. 30A) Bar graph showing RT-qPCR quantification of expression levels of collagen I (Coll), collagen III (Col3). and a-smooth muscle actin (a-SMA) in cultured facial fibroblasts and dorsal fibroblasts receiving SLIT2 treatment (SLIT2+) or vehicle control (SLIT2-).FIG. 30B) ELISA quantification of collagen 1 and TGF-betal levels in the conditioned media of cultured facial fibroblasts and dorsal fibroblasts receiving SLIT2 treatment (SLIT2+) or vehicle control (SLIT2-). For all of the above experiments, three independent fibroblast cultures from three different groups of mice were used. Unless otherwise noted, between-group statistical differences were assessed using one-way ANOVA with post-hoc Tukey HSD test (ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001,).

[0053] FIGS . 31 A-31 D illustrate that SLIT2 treatment reduces scarring in dorsal wounds. FIG. 31 A) IHC staining and quantification of staining intensity (bar graph) of ROBO2, COL1, and SOX10 in healed wounds at POD 14; nuclear staining (DAPI) (scale bar, 120pm). FIG. 31B) Expression levels of fibrosis markers (Coll, Col3, aSMA) by RT-qPCR in healed wounds at POD 14. FIG. 31C) ELIS A quantification of collagen 1 levels in the wound lysate of experimental groups. FIG. 3 ID) Diagram of SLIT2’s role in dermal fibrosis. Between-group statistical differences were assessed using one-way ANOVA with post-hoc Tukey HSD test (ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0054] FIGS. 32A-32C illustrate that CRISPR-mediated Eidl knockout and overexpression in fibroblasts and injection into dorsal wounds. FIG. 32A) Low-magnification images of picrosinus red staining of post-operative day- 14 (POD- 14) untreated facial wounds (FW), untreated dorsal wounds (DW), dorsal wounds injected with knockout-control fibroblasts (DW KO Ctrl), dorsal wounds injected with Eidl -knockout fibroblasts (DW Eidl KO), dorsal wounds injected with overexpression-control fibroblasts (DW OE Ctrl), and dorsal wounds injected with Eidl- overexpressing fibroblasts (DW Eidl OE); wounded areas bounded by dotted lines (FW and DW Eidl OE scale bars, 15pm; all other scale bars, 100pm). FIG. 32B) Bar graph showing RT-qPCR quantification of expression levels of collagen I. collagen III, and otSMA and FIG. 32C) ELISA of collagen levels in wound lysate of POD- 14 FW, DW. DW KO Ctrl. DW Eidl KO, DW OE Ctrl, and DW Eidl OE. Between-group statistical differences were assessed using one-way ANOVA with post-hoc Tukey7HSD test (ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0055] FIGS. 33Aand 33B illustrate that in vitro small-molecule inhibition of EP300 attenuates scarring phenotype. FIG. 33A) ELISA quantification of COL1 levels in the media of cultured facial fibroblasts, dorsal fibroblasts, and EP300i-treated dorsal fibroblasts. FIG. 33F) Fluorescence- activated cell sorting (FACS) of cultured facial fibroblasts, dorsal fibroblasts, and EP300i-treated dorsal fibroblasts, showing percentage of fibroblasts positive for SOX10. Unless otherwise noted, between-group statistical differences were assessed using one-way ANOVA with post-hoc Tukey HSD test (ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0056] FIGS. 34 A and 34B illustrate that in vivo EP300 inhibition promotes facial-like healing with reduced fibrosis. FIG. 34A) Expression levels of fibrosis markers Coll, Col3, aSMA) in POD- 14 wounds of the experimental groups. FIG. 34B) ELISA quantification of collagen 1 levels in the wound lysate of the experimental groups. Unless otherwise noted, between-group statistical differences were assessed using one-way ANO VA with post-hoc Tukey HSD test (ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

[0057] FIGS. 35A-35C illustrate that in vivo Ep300 knockout promotes facial-like dorsal w ound healing with reduced fibrosis. FIG. 35 A) H&E staining of healed w ounds at POD 14 of the different experimental groups, with scars bounded by dotted lines and quantification of scar width and follicle number below (scale bar, 150pm). FIG. 35B) UMAP of quantified collagen fiber features based on picrosirius red collagen staining for each experimental group at POD 14. FIG. 35C) Fluorescence imaging of GFP levels and IHC of SOX10 in healed wounds at POD 14 for each experimental group; nuclear staining (DAPI) (scale bar, 120pm). Between-group statistical differences were assessed using one-way ANOVA with post-hoc Tukey HSD test (*p < 0.05, **p < 0.01, ****p < 0.000i).

[0058] FIGS. 36A-36D illustrate single-cell RNA sequencing of facial and dorsal wounds after EP300 inhibition. FIG. 36A) Uniform manifold approximation and projection (UMAP) plot of fine Seurat clustering of all sequenced cells from the scRNA-seq experiment described in FIG. 5D. FIG. 36B) UMAP of all sequenced cells by cell type from the scRNA-seq experiment described in FIG. 5D. FIG. 36C) UMAP of all sequenced cells by cell type in each experimental group from scRNA- seq in FIG. 5D. FIG. 36D) Left: Schematic demonstrating anchor label transfer analysis of FACS- isolated fibroblasts (from FIGS. 2A-2C) onto fibroblast clusters from the EP300i experiment (from FIG. 5D). Right: Feature plots of FACS-isolated fibroblasts (clusters 0-5) mapped onto fibroblasts from the EP300i experiment (clusters 1-9).

[0059] FIGS. 37A-37D illustrate a single cell assay for transposase-accessible chromatin of facial and dorsal wounds after EP300 inhibition. FIG. 37 A) Schematic of experiment showing facial unwounded skin (FUW), facial wounds treated with PBS (FW PBS), facial wounds treated with EP300 inhibitor (FW EP300i), dorsal unwounded skin (DUW), dorsal wounds treated with PBS (DW PBS), and dorsal treated wounds with EP300 inhibitor (DW EP300i). at post-operative day 14 (POD 14) for single-cell assay for transposase-accessible chromatin (scATAC-seq) analysis. FIG. 37B) Uniform manifold approximation and projection (UMAP) plot of fine Seurat clustering of all ATAC-sequenced cells colored by Seurat group. FIG. 37C) UMAP of all ATAC-sequenced cells colored by cell type. FIG. 37D) UMAP of all ATAC-sequenced cells by cell type by group.

[0060] FIGS. 38A-38D illustrate fibroblast subpopulations and chromatin accessibility changes by single cell assay for transposase-accessible chromatin sequencing in facial and dorsal wounds after EP300 inhibition. FIG. 38A) Uniform manifold approximation and projection (UMAP) of fibroblast clusters in all sequenced fibroblasts from single cell sequencing assay for transposase-accessible chromatin (scATAC-seq). FIG. 38B) UMAP of ATAC-sequenced fibroblasts from facial unwounded skin (FUW), facial wounds treated with PBS (FW PBS), facial wounds treated with EP300 inhibitor (FW EP300i), dorsal unwounded skin (DUW), dorsal wounds treated with PBS (DW PBS), and dorsal wounds treated with EP300 inhibitor (DW EP300i), analyzed at post-operative day 14 (POD 14). FIG. 38C) Pie charts showing proportions of each fibroblast scATAC-seq cluster in different experimental groups. FIG. 38D) Gene Ontology pathway analysis of scATAC-seq fibroblast clusters 0. 3, 4, and 5.

[0061] FIGS. 39A-39B illustrate CODEX analysis of facial and dorsal wounds. FIG. 39A) Representative images of the distribution of CODEX-annotated cell populations from facial unwounded skin (FUW), facial wounds treated with PBS (FW PBS), facial wounds treated with EP300 inhibitor (FW EP300i), dorsal unwounded skin (DUW), dorsal wounds treated with PBS (DW PBS), and dorsal wounds treated with EP300 inhibitor (DW EP300i). analyzed at post-operative day 14 (POD 14). FIG. 39B) Ridge plots of CODEX panel fibroblast marker and ECM protein levels (intensity; x-axis) in each CODEX-generated fibroblast cluster (y- axis): FBI -5 denote CODEX fibroblast cluster 1-5.DETAILED DESCRIPTION OF EMBODIMENTS

[0062] “EP300 modulator’ as defined herein is a molecule (e.g. polypeptide, small molecule or other biomolecule) that modulates EP300 activity, directly or indirectly, through association or inhibition of biological activity at any point in the EP300 pathway. In some embodiments, an EP300 modulator is an amino acid sequence. In some embodiments, an EP300 modulator is an amino acid sequence or polypeptide that activates or associates with ROBO2. activates or associates to SL1T2, or activates or associates to E1D1. In some embodiments, an EP300 modulator is a small molecule. In some embodiments, an EP300 modulator is an inhibitor of EP300 or associates to EP300 and disrupts its wild ty pe biological activity. In some embodiments, an EP300 modulator is an amino acid sequence or polypeptide that activates ROBO2. activates SLIT2, or activates EID1, or other polypeptides in the EP300 pathway. In some embodiments, an EP300 modulator is an inhibitor of an acetyltransferase inthe EP300 pathway. In some embodiments, an EP300 modulator is an inhibitor of EP300 acetyltrasferase.

[0063] As used herein in reference to a chemical structure, a derivative, also known as a chemical derivative, is a compound having a structure similar to that of a referenced compound, but differing from it in respect to a certain component. “Derivatives” of compounds disclosed herein are pharmaceutically acceptable salts, prodrugs, deuterated forms, radio-actively labeled forms, isomers, tautomers, solvates and combinations thereof. The “combinations” mentioned in this context are refer to derivatives falling within at least two of the groups: pharmaceutically acceptable salts, prodrugs, deuterated forms, radio-actively labeled forms, isomers, tautomers, and solvates. Examples of radio-actively labeled forms include compounds labeled with radioactive moieties; for example, tritium, phosphorous-32, iodine-129. carbon-11, and fluorine- 18.

[0064] The terms “pharmaceutically acceptable salt” includes pharmaceutically acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include e.g.. salts of inorganic acids (such as hydrochloric acid, hydrobromic, phosphoric, nitric, and sulfuric acids) and of organic acids (such as, acetic acid, benzenesulfonic, benzoic, methanesulfonic, and p-toluenesulfonic acids). Examples of pharmaceutically acceptable base addition salts include e.g., sodium, potassium, calcium, ammonium, organic amino, or magnesium salt.

[0065] The term “pharmaceutically acceptable carrier” refers to a non-toxic earner, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene- polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0066] As used herein, the phrase “pharmaceutically acceptable” means those compounds, materials, compositions, and / or dosage forms which are. within the scope of sound medical judgment, suitable for use in contact with tissues of humans and animals. In some embodiments, “pharmaceutically acceptable” means approved by a regulatory agency of theFederal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0067] In some embodiments, pharmaceutical compositions comprise one or more compound, analog, or derivative herein in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0068] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity' can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0069] These pharmaceutical compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0070] In some embodiments, in order to prolong the effect of a compound herein, the absorption of the drug is slowed. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0001] As used in reference to an compound herein, an “analog” refers to a substituted version of the compound. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In some embodiments, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and nonaromatic substituents of organic compounds. Exemplarysubstituents include those described below. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Also, the terms ‘‘substitution” or “substituted with” include the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound; e.g.. a compound that does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. It is also contemplated that, in certain embodiments, unless expressly indicated to the contrary, individual substituents can be further optionally substituted (i.e., further substituted or unsubstituted).

[0002] In defining various terms; for example, “R1,” “R2,” R3,” “R4,” “A1,” “A2,” “A3,” and “A4,” are used herein as generic symbols to represent various specific substituents. These symbols can be any substituent, not limited to those disclosed herein, and when they are defined to be certain substituents in one instance, they can, in another instance, be defined as some other substituents.

[0003] In some embodiments, a substituent includes a “halo” or “halogen.” The terms “halo” and “halogen” as used herein refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo. -Br), and iodine (iodo, -I).

[0004] In some embodiments, a substituent includes an “alkyl” group. The term “alkyl,” as used herein, refers to a monovalent saturated, straight- or branched-chain hydrocarbon radical, having unless otherwise specified, 1-6 carbon atoms. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, n-pentyl, tert-pentyl, neopentyl, sec-pentyl, 3-pentyl, sec-isopentyl, hexyl, 2-methylpentane, 3- methylpentane, 2,2-dimethylbutane, 2,3-dimentybutane and the like.

[0005] In some embodiments, a substituent includes a "haloalkyl" group. The term “haloalkyl” includes mono, poly, and perhaloalkyl groups where the halogens are independently selected from fluorine, chlorine, bromine, and iodine.

[0006] In some embodiments, a substituent includes an “alkoxy” group. “Alkoxy” is an alkyl group which is attached to another moiety via an oxygen linker (-O(alkyl)). Non-limiting examples include methoxy, ethoxy, propoxy, and butoxy.

[0007] In some embodiments, a substituent includes a “haloalkoxy” group. “Haloalkoxy” is a haloalkyl group which is attached to another moiety via an oxygen atom such as, e.g., but are not limited to -OCHCF2 or -OCFv

[0008] In some embodiments, a substituent includes a “9- to 10-membered carbocyclyl” group. The term “9- to 10-membered carbocyclyF’ means a 9- or 10- membered monocyclic, bicyclic (e.g., a bridged or spiro bicyclic ring), polycyclic (e.g., tricyclic), or fused hydrocarbon ring system that is saturated or partially unsaturated. The term “9- to 10-membered carbocyclyl” also includes saturated or partially unsaturated hydrocarbon rings that are fused to one or more aromatic or partically saturated hydrocarbon rings (e.g.. dihydroindenyl and tetrahydronaphthalenyl). Bridged bicyclic cycloalkyl groups include, without limitation. bicyclo[4.3.1]decanyl and the like. Spiro bicyclic cycloalkyl groups include, e.g., spiro[3.6]decanyl, spiro[4.5]decanyl, spiro [4.4]nonyl and the like. Fused cycloalkyl rings include, e.g., decahydronaphthalenyl, dihydroindenyl, decahydroazulenyl, octahydroazulenyl, tetrahydronaphthalenyl, and the like. It will be understood that when specified, optional substituents on a carbocyclyl (e.g., in the case of an optionally substituted cycloalkyl) may be present on any substitutable position and, include, e.g., the position at which the carbocyclyl group is attached.

[0009] In some embodiments, a substituent includes a “cycloalkyl” group. A cycloalkyl is a completely saturated carbocycle and includes e.g.. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0010] In some embodiments, a substituent includes a “9-membered fused heterocyclyl” group. The term “9-membered fused heterocyclyl” means a 9-membered saturated or partially unsaturated fused monocyclic heterocyclic ring comprising at least one oxygen heteroatom and optionally two to four additional heteroatoms independently selected from N, O, and S. The terms “heterocycle,” “heterocyclyl,” “heterocyclyl ring,” “heterocyclic group,” “heterocyclic moiety.” and “heterocyclic radical,” are used interchangeably herein. A heterocyclyl ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. Examples of fused saturated or partially unsaturated heterocyclic radicals compristing at least one oxygen atom include, without limitation, dihydrobenzofuranyl, dihydrofuropyridinyl, octahydrobenzofuranyl, and the like. Where specified as being optionally substituted, substituents on a heterocyclyl (e.g., in the case of an optionally substituted heterocyclyl) may be present on any substitutable position and include, e.g, the position at which the heterocyclyl group is attached.

[0011] In some embodiments, a substituent includes a “5- or 6- membered heteroaryl" group. The term “5- or 6- membered heteroaryl” refers to a 5- or 6-membered aromatic radical containing 1-4 heteroatoms selected from N, O. and S. Nonlimiting examples include thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl,thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, etc. When specified, optional substituents on a heteroaryl group may be present on any substitutable position and, include, e.g., the position at which the heteroaryl is attached.

[0071] As described herein, compounds of the disclosure may contain '‘optionally substituted” moieties. In general, the term “substituted,” whether preceded by the term “optionally” or not, means that one or more hydrogens of the designated moiety are replaced wi th a suitable substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds. In is also contemplated that, in certain embodiments, unless expressly indicated to the contrary, individual substituents can be further optionally substituted (z.e., further substituted or unsubstituted).

[0072] In some embodiments, an analog retains at least about 10, about 15, about 20, about 25. about 30. about 40, about 45, about 50. about 55. about 60. about 65, about 70, about 75. about 80, about 85, about 90, about 91, about 92, about 93, about 94, about 95, about 96, about 97, about 98, about 99, or about 100% of the activity of the referenced compound. Assays for the activity of a compound are found in the examples herein.

[0073] It can differ in one or more atoms, functional groups, or substructures, which are replaced with other atoms, groups, or substructures. A structural analog can be imagined to be formed, at least theoretically, from the other compound. Structural analogs are often isoelectronic.

[0074] The term “amino acid” refers to a molecule containing both an amino group and a carboxyl group bound to a carbon which is designated the a-carbon. Suitable amino acids herein include, without limitation, both the D- and L-isomers of the naturally -occurring amino acids, as well as non-naturally occurring amino acids prepared by organic synthesis (synthetic amino acids) or other metabolic routes. Amino acid, as used herein, also encompasses non- proteogenic amino acids, beta amino acids, homo-amino acids, beta-homo-ammo acids, amino acids with modified side groups, N-methyl amino acids, alpha-methyl amino acids and D- amino acids. In some embodiments, a single “amino acid” might have multiple sidechain moieties, as available per an extended aliphatic or aromatic backbone scaffold. Unless the context specifically indicates otherwise, the term amino acid, as used herein, is intended to include amino acid analogs or derivatives.

[0075] The term “amino acid sequence” as used herein means a compound composed of at least two constituent amino acids connected by covalent bonds. In some embodiments, the connecting covalent bond is a peptide bond. The constituent amino acids may be from the group of the amino acids encoded by the genetic code and they may be natural amino acids which are not encoded by the genetic code, as well as synthetic amino acids. Natural amino acids which are not encoded by the genetic code are e.g, " / -carboxy glutamate, ornithine, phosphoserine, D- alanine and D-glutamine. Synthetic amino acids comprise amino acids manufactured by chemical synthesis, and in some embodiments are, for example, D-isomers of the amino acids encoded by the genetic code (for example, D-alanine and D-leucine), Aib (a-aminoisobutyric acid), Abu (a-aminobutyric acid), Tie (tert-but l glycine). P-alanine, 3-aminomethyl benzoic acid, and anthranilic acid.

[0076] A “non-essential” amino acid residue is a residue that can be altered from the wildtype sequence of an amino acid sequence without abolishing or substantially altering its essential biological or biochemical activity (e.g., receptor binding or activation). An “essential” amino acid residue is a residue that, when altered from the wild-type sequence of the polypeptide, results in abolishing or substantially abolishing the polypeptide’s essential biological or biochemical activity.

[0077] A “non-natural side chain” is a modified or synthetic chain of atoms joined by covalent bond to the a-carbon atom, p-carbon atom, or y-carbon atom which does not make up the backbone of the polypeptide chain of amino acids. For example, the natural side chain, or R group, of alanine is a methyl group, and anon-natural side chain of alanine may include one or more deuterium in place of the natural hydrogen(s).

[0078] A “non-naturally encoded amino acid” refers to an amino acid that is not one of the 20 common amino acids or pyrolysine or selenocysteine. Other terms that may be used synonymously with the term “non-naturally encoded amino acid” are “non-natural amino acid,” “unnatural amino acid,” “non-naturally-occurring amino acid,” and variously hyphenated and non-hyphenated versions thereof. The term “non-naturally encoded amino acid” also includes, but is not limited to, amino acids that occur by modification (e.g., post- translational modifications) of a naturally encoded amino acid (including but not limited to, the 20 common amino acids or pyrolysine and seienocysteine) but are not themselves naturally incorporated into a growing polypeptide chain by the translation complex. Examples of non- naturally -occurring amino acids include, but are not limited to, N-acetylglucosaminyl-L -serine N acetylglucosaminyl-L-threonine and O phosphotx rosine.

[0079] An “amino terminus modification group” refers to any molecule that can be attached to the amino terminus of an amino acid sequence. Similarly, a “carboxy terminus modification group” refers to any molecule that can be attached to the carboxy terminus of an amino acid sequence. Terminus modification groups include, but are not limited to, various water soluble polymers, peptides or proteins such as serum albumin, immunoglobulin constant region portions such as Fc, or other moieties that increase serum half-life of peptides. Amidation is a terminus modification groups, including a carboxy terminus modification. In some embodiments, an amino acid sequence herein, an amino acid sequence in a composition herein, or an amino acid sequence in a pharmaceutical composition herein includes a carboxyterminal amidation as a modification.The term “linkage” or “linker” refers to a structure that connects two or more additional structures Examples of linkers include peptide linkers, protein linkers, PEG linkers, and combinations thereof. A "maleimide-PEG linker", as used herein, refers to a chemical moiety comprising a polyethylene glycol (PEG) polymer of the formula "-(O-CFh-CEh)]!-" wherein "n” is from about 3 through about 24, and a derivatized maleimide functional group, wherein said linker may form a covalent attachment to an amino acid sequence disclosed herein through a thioether bond between a maleimide functional group and a cysteine residue in the antibody or an antigen-binding fragment , and / or or may form a covalent atachment to an N-formyl- niethionine peptide through an amide bond to the epsilon amino side chain of the C-terminal lysine of a N-fonnyl-methionine peptide or an amide bond to the gamma carboxyl group of the C-terminal glutamic acid of a N-fonnyl-methionine peptide.. Hydrolytically stable linkages means that the linkages are substantially stable in water and do not react with water at useful pH values, including but not limited to, under physiological conditions for an extended period of time, perhaps even indefinitely. Hydrolytically unstable or degradable linkages mean that the linkages are degradable in water or in aqueous solutions, including for example, blood. Enzymatically unstable or degradable linkages mean that the linkage can be degraded by one or more enzymes. As understood in the art, PEG and related polymers may include degradable linkages in the polymer backbone or in the linker group between the polymer backbone and one or more of the terminal functional groups of the polymer molecule. For example, ester linkages formed by the reaction of PEG carboxy lie acids or activated PEG carboxy lie acids with alcohol groups on a biologically active agent generally hydrolyze under physiological conditions to release the agent. Other hydrolytically degradable linkages include, but are not limited to, carbonate linkages; imine linkages resulted from reaction of an amine and analdehyde; phosphate ester linkages formed by reacting an alcohol with a phosphate group; hydrazone linkages which are reaction product of a hydrazide and an aldehyde; acetal linkages that are the reaction product of an aldehyde and an alcohol; orthoester linkages that are the reaction product of a formate and an alcohol; peptide linkages formed by an amine group, including but not limited to, at an end of a polymer such as PEG, and a carboxyl group of a peptide; and oligonucleotide linkages formed by a phosphoramidite group, including but not limited to, at the end of a polymer, and a 5' hydroxyl group of an oligonucleotide. In some embodiments, a linker herein is gamma-Gly or GABA.

[0080] The term “polypeptide” encompasses two or more naturally or non-naturally- occurring amino acids joined by a covalent bond (e.g., an amide bond). Polypeptides as described herein include full-length proteins (e.g., fully processed pro-proteins or full-length synthetic polypeptides) as well as shorter amino acid sequences (e.g., fragments of naturally- occurring proteins or synthetic polypeptide fragments).

[0081] A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., K, R, H), acidic side chains (e.g., D, E), uncharged polar side chains (e.g., G, N, Q, S, T, Y, C, H), nonpolar side chains (e.g., G, A, V, L, I, P, F, M, W), betabranched side chains (e.g. , T. V, I) and aromatic side chains (e.g., Y, F, W, H). Thus, a predicted nonessential amino acid residue in an amino acid sequence, for example, replaced with another amino acid residue from the same side chain family. Other examples of conserved amino acid substitutions are substitutions based on isosteric considerations (e.g., norleucine for methionine) or other properties (e.g., 2-thienylalanine for phenylalanine).

[0082] As used herein, the term “derived from” in the context of the relationship between a chemical structure or amino acid sequence and a related chemical structure or related amino acid sequence describes a chemical structure homologous or structurally related to a reference chemical structure, or a biological sequence (e.g., an amino acid sequence) based on but varying from a reference biological sequence (e.g., by amino acid change or chemical modification).

[0083] The term “functional variant thereof’ refers to a peptide varying from the reference sequence but retaining biological activity compared to a peptide having the reference sequence. In some embodiments, the biological activity is measured by a functional assay disclosed herein. In some embodiments, a “functional variant thereof’ has at least about 70%. 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity, or asequence identify from about any two of 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%. 96%. 97%. 98%. 99%. or 100%, or a sequence identity of about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% to a stated reference sequence. In some embodiments, a functional variant of an amino acid sequence herein retains at least about 10, about 15, about 20, about 25, about 30, about 40, about 45, about 50, about 55, about 60, about 65. about 70. about 75, about 80, about 85, about 90, about 91, about 92, about 93, about 94. about 95. about 96, about 97, about 98, about 99, or about 100% of the biological activity of the reference sequence from which it is a variant.

[0084] “Variants’’ are intended to mean similar sequences, and any solvates, salts, or mutants of a wild-type or reference sequence upon which the variant is based. For nucleic acid molecules, a variant comprises a nucleic acid molecule having deletions (i.e.. truncations) at the 5’ and / or 3’ end; deletion and / or addition of one or more nucleotides at one or more internal sites in the native polynucleotide and / or substitution of one or more nucleotides at one or more sites in the native polynucleotide. For amino acid sequences, the variant is any mutant sequence of a reference sequence including any one or combination of an amino acid sequence: that is a truncation mutant, that has an addition, that has a deletion. As used herein, a “native” nucleic acid molecule or polypeptide sequence comprises a naturally occurring or endogenous nucleotide sequence or amino acid sequence, respectively. For nucleic acid molecules, conservative variants include those sequences that, because of the degeneracy of the genetic code, encode the amino acid sequence of one of the polypeptides of the disclosure. Variant nucleic acid molecules and amino acid sequences also include synthetically derived nucleic acid molecules, such as those generated, for example, by using site-directed mutagenesis but which still encode a protein of the disclosure. Generally, variants of a particular nucleic acid molecule of the disclosure will have at least about 70%. 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to an amino acid sequence disclosed herein. Variants of a particular nucleic acid molecule of the disclosure (i.e., the reference DNA sequence) can also be evaluated by comparison of the percent sequence identity between the amino acid sequence encoded by a variant nucleic acid molecule and the amino acid sequence encoded by the reference nucleic acid molecule. Percent sequence identity between any two amino acid sequences can be calculated using sequence alignment programs and parameters described elsewhere herein. In some embodiments, the term “variant” amino acid sequence is intended to mean an amino acid sequence derived from the native amino acid sequence by deletion (so-called truncation) of one or more amino acids at the N-terminal and / or C-terminal end of the native amino acid sequence; deletion and / or addition of one or more amino acids atone or more internal sites in the native amino acid sequence; or substitution of one or more amino acids at one or more sites in the native amino acid sequence. Amino acid sequences encompassed by the present disclosure are biologically active, that is they are functional variants, as above, and continue to possess the desired biological activity of the native amino acid sequences as described herein. Biologically active variant is synonymous with functional variant herein. Such variants may result from, for example, genetic polymorphism or from human manipulation. In some embodiments, functional variants of an amino acid sequence of the disclosure will have at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity' to the amino acid sequence for the native amino acid sequence as determined by sequence alignment programs and parameters described elsewhere herein. A biologically active variant of an amino acid sequence of the disclosure may differ from that amino acid sequence by from about 1 to about 10 amino acid residues, as few as about 1 to about 15, for example from about 6 to about 10, as few as about 5, as few as about 4, 3, 2, or even 1 amino acid residue. The amino acid sequences of the disclosure may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions. Methods for such manipulations are generally known in the art. For example, amino acid sequence variants and fragments of the proteins or amino acid sequences can be prepared by mutations in the nucleic acid sequences that encode the proteins or amino acid sequences recombinantly.

[0085] Percent identity refers to the number of aligned positions between a sequence and a reference sequence where the matching characters (e.g., amino acids in proteins or nucleotides in nucleic acids) are identical divided by the length of the reference sequence, and where the quotient is then multiplied by one hundred. The “percent identity'’ of two polynucleotide or two amino acid sequences is determined by comparing the sequences using the GAP computer program (a part of the GCG Wisconsin Package, version 10.3 (Accelrys, San Diego, Calif.)) using its default parameters. “Identical” or “identity” as used herein in the context of two or more nucleic acids or amino acid sequences, may mean that the sequences have a specified percentage of residues that are the same over a specified region. The percentage may be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified regionof comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) may be considered equivalent. Identity may be performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0. Briefly, the BLAST algorithm, which stands for Basic Local Alignment Search Tool is suitable for determining sequence similarity. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov). This algorithm involves first identifying high scoring sequence pair (HSPs) by identifying short words of length Win the query' sequence that either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Extension for the word hits in each direction are halted when: 1) the cumulative alignment score falls off by the quantify X from its maximum achieved value; 2) the cumulative score goes to zero or below, due to the accumulation of one or more negativescoring residue alignments; or 3) the end of either sequence is reached. The Blast algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The Blast program uses as defaults a word length (W) of 11, the BLOSUM62 scoring matrix (see Henikoff et al., Proc. Natl. Acad. Sci. USA, 1992, 89, 10915-10919, which is incorporated herein by reference in its entirety ) alignments (B) of 50, expectation (E) of 10, M=5, N=4, and a comparison of both strands. The BLAST algorithm (Karlin et al., Proc. Natl. Acad. Sci. USA, 1993, 90, 5873- 5787, which is incorporated herein by reference in its entirety) and Gapped BLAST perform a statistical analysis of the similarity between two sequences. One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two sequences would occur by7chance. For example, a nucleic acid is considered similar to another if the smallest sum probability7in comparison of the test nucleic acid to the other nucleic acid is less than about 1, less than about 0.1, less than about 0.01, and less than about 0.001. Two single-stranded polynucleotides are “the complement” of each other if their sequences can be aligned in an anti-parallel orientation such that every7nucleotide in one polynucleotide is opposite its complementary7nucleotide in the other polynucleotide, w ithout the introduction of gaps, and without unpaired nucleotides at the 5?or the 3’ end of either sequence. A polynucleotide is “complementary” to another polynucleotide if the two polynucleotides can hybridize to one another under moderatelystringent conditions. Thus, a polynucleotide can be complementary to another polynucleotide without being its complement.

[0086] The term '‘derivative” as used herein in relation to an amino acid sequence means a chemically modified amino acid sequence or a variant thereof, wherein at least one substituent is not present in the unmodified amino acid sequence or a variant thereof; i.e., a peptide which has been covalently modified. Typical modifications are amides, carbohydrates, alkyl groups, acyl groups, esters and the like. The term “derivative” also encompasses a protein, peptide, or amino acid sequence including one or more non-naturally encoded amino acid.”

[0087] The term “fragment” refers to any variant of a naturally occurring amino acid disclosed herein that comprises at least 4 amino acids identical to the naturally occurring amino acid sequence upon which the variant is based. The term “functional fragment” refers to any fragment of any variant of a naturally occurring amino acid sequence disclosed herein that comprises at least 4 amino acids identical to the naturally occurring amino acid sequence upon which the variant is based and shares the function of the naturally occurring polypeptide upon which the variant is based. In some embodiments, the fragment comprises at least 4, 5, 6, 7, 8, 9, 10. 11. 12. 13. 14, 15, 16, 17, 18, 19. or 20 amino acids common with the naturally occurring peptide.

[0088] “Effective amount” refers to an amount of an amino acid sequence, a composition, or pharmaceutical composition as described herein effective to achieve a particular biological result such as, but not limited to. biological results disclosed, described, or exemplified herein. Such results may include, but are not limited to, the effective reduction of symptoms associated with any of the disease states mentioned herein, as determined by any means suitable in the art. The effective amount of the composition may be dependent on any number of variables, including without limitation, the species, breed, size, height, weight, age. overall health of the subject, the type of formulation, the mode or manner or administration, the type and / or severity of the particular condition being treated, or the need to modulate the activity of the molecular pathw ay induced by association of the analog to its receptor. The appropriate effective amount can be routinely determined by those of skill in the art using routine optimization techniques and the skilled and informed judgment of the practitioner and other factors evident to those skilled in the art. A therapeutically effective dose of variants or derivatives described herein may provide partial or complete biological activity as compared to the biological activity induced by the wild-type or naturally occurring amino acid sequences upon which the variants or derivatives are derived. A therapeutically effective dose of the variants or derivatives described herein may provide a sustained biochemical or biological affect and / or an increasedresistance to degradation when placed in solution as compared with the normal affect observed when the naturally occurring and fully processed and translated protein or peptide is administered to the same subject.

[0089] The term “biological activity” encompasses structural and functional properties of an amino acid sequence herein. In some embodiments, biological activity is, for example, inhibiting EP300. In some embodiments, biological activity is, for example, inhibiting expression of EP300. In some embodiments, biological activity is. for example, increasing expression of SLIT2. In some embodiments, biological activity is, for example, increasing activity of SLIT2. In some embodiments, biological activity is, for example, increasing expression of EIDl. In some embodiments, biological activity is, for example, increasing activity of EIDl. In some embodiments, biological activity’ is, for example, increasing expression of ROBO2. In some embodiments, biological activity is, for example, increasing activity7of ROBO2.

[0090] Unless defined otherwise, 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. For example, Singleton et al.. Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, NY 1994), provide one skilled in the art with a general guide to many of the terms used in the present application. Additionally, the practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, and biochemistry, which are within the skill of the art. Such techniques are explained fully in the literature, such as, "Molecular Cloning: A Laboratory Manual", 2nd edition (Sambrook et al., 1989); "Oligonucleotide Synthesis" (M.J. Gait, ed., 1984); "Animal Cell Culture" (R.I. Freshney, ed., 1987); "Methods in Enzymology" (Academic Press, Inc.); "Handbook of Experimental Immunology", 4th edition (D.M. Weir & C.C. Blackwell, eds., Blackwell Science Inc., 1987); "Gene Transfer Vectors for Mammalian Cells" (J.M. Miller & M.P. Calos, eds., 1987); "Current Protocols in Molecular Biology" (F.M. Ausubel et al., eds., 1987); and "PCR: The Polymerase Chain Reaction". (Mullis et al., eds.. 1994).

[0091] As used in the present disclosure and claims, the singular forms “a”, “an” and “the” include plural forms unless the context clearly dictates otherwise.

[0092] It is understood that wherever embodiments are described herein with the language “comprising” otherwise analogous embodiments described in terms of “consisting of’ and / or “consisting essentially of’ are also provided. It is also understood that wherever embodimentsare described herein with the language “consisting essentially of’ otherwise analogous embodiments described in terms of “consisting of’ are also provided.

[0093] The term "about" as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20%, ±10%, ±5%„ ±1%, or ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. For recitation of numeric ranges herein, each intervening number therebetween with the same degree of precision is explicitly contemplated. For example, for the range of from about 6 to about 9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6,9, and 7.0 are explicitly contemplated.

[0094] The term “and / or” as used in a phrase such as “A and / or B” herein is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term “and / or” as used in aphrase such as “A, B, and / or C” is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0095] The term “exposing” as used herein refers to bringing a disclosed compound, biomolecule, a cell, target receptor, or other biological entity in close proximity’ for direct or indirect contact, in such a manner that the compound, biomolecule, cell, receptor, or first biological entity can affect the activity of a second biological entity (e.g, receptor, cell, etc.). This can occur directly by physical contact between the disclosed compound and a disclosed cell, such as a fibroblast disclosed herein, receptor or other entity; i.e., by interacting with the target or cell itself. Exposing can occur indirectly by interacting by one biological entity (biomolecule, cell or reagent) being exposed to another molecule, co-factor, factor, or protein on which the activity of the second biological entity is dependent. In some embodiments, the activity of a cell (a second biological entity) in response to the compound or molecule (the first biological entity) is differentiation. In some embodiments, the compound is one or more differentiation factors. In some embodiments, the differentiation factor or factors are a component of cell culture medium within which fibroblasts are cultured.

[0096] The term “substantially free of’ as used herein refers to a composition that only has trace or negligible amounts of the substance to which it refers. In some embodiments, substantially free means that the composition comprises only about 0%, 0.1%, 0.2%, 0.3% 0.4% or 0.5% of the substance to which it refers. In some embodiments, substantially free means that the composition comprises less than about 1.0% of the substance to which it refersrelative to the number or mass of substances in the compositions and confers no biological effect to the compositions.Compositions

[0097] We provide, for the first time, evidence that facial fibroblasts harbor a reduced fibrotic potential derived from their neural crest origins. We show that compared to fibroblasts from other anatomical locations, facial fibroblasts upregulate SLIT2-ROBO2 signaling, a pathway important in neural crest cell biology,23'25to promote wound healing with less fibrosis. We found that SLIT2-ROBO2 acts through EID 1 -mediated inhibition of EP300 to maintain chromatin in a more transcriptionally silent state (FIG. 1A). Genetic and small molecule manipulation of the SLIT2-ROBO2-EID1 axis in fibroblasts allowed dorsal wounds to heal like facial wounds with reduced scarring, demonstrating an ability to harness the inherent plasticity of adult fibroblasts for anti-fibrotic therapies.

[0098] The disclosure relates to a composition comprising one or more EP300 modulator, and optionally a pharmaceutically acceptable carrier. The disclosure also relates to a methods of using the aforementioned compositions for use in treatment of facial wounds of a subject and in methods of accelerating wound healing of a subject. In some embodiments, the compositions are administered to the subject topically, intraperitoneally, intradermally or subdermally.

[0099] In some embodiments, the disclosure relates to an amino acid sequence, a nucleic acid sequence encoding the amino acid sequence, and compositions or pharmaceutical compositions comprising one or both of the amino acid sequence or the nucleic acid sequence. In some embodiments, the amino acid sequence, the nucleic acid sequence encoding the amino acid sequence, or the compositions or pharmaceutical compositions comprising one or both of the amino acid sequence or the nucleic acid sequence encodes an EP300 modulator. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of one or more of the amino acid sequence the nucleic acid sequence. In some embodiments, the amino acid sequence is one from Table 1 or a functional variant thereof. In some embodiments, the compositions comprise a nucleic acid sequence encoding an amino acid sequence of Table 1 or a functional variant thereof. In some embodiments, the composition or pharmaceutical composition comprises a plurality of amino acid sequences of Table 1 or a functional variant of one or more of the plurality of amino acid sequences. In some embodiments, the composition or pharmaceutical composition comprises a plurality of nucleic acid sequences encoding amino acid sequences of Table 1 or a functional variant of one or more of the plurality of nucleic acidsequences. In some embodiments, the composition or pharmaceutical composition comprises at least one amino acid sequence of Table 1 or a functional variant thereof and at least one of nucleic acid sequences encoding an amino acid sequences of Table 1 or a functional variant thereof.Table 1

[0100] In some embodiments, the amino acid sequence comprises MAEMAELCELYEESNELQMDVLPGEGYMEVGRGARGPAPEEGPMEEEAGPAAAR AQRGLFPEAGADLEGDEFDDWEDDYEFPEEERWSGAMHRVSAALEEANKVFLRT ARAGDALDGGFQARCEKSPFDQLAFIEELFSLMVVNRLTEELGCDEIIDRE (SEQ ID NO: 1), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 1. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 1. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 1 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, or about 158 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5. about 6, about 7. about 8, about 9. about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130,about 140, or about 150 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, or about 158 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 1 from which it was derived.

[0101] In some embodiments, the amino acid sequence comprises DLEGDEFDDWEDDYE (SEQ ID NO: 2), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 2. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 2. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 2 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, or about 14 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5, about 6. about 7, about 8. about 9, about 10, about 11, about 12, or about 13 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 11, about 12, about 13, or about 14 amino acids. In some embodiments, the functional fragment has at least about 75%. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%. 97%. 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 2 from which it was derived.

[0102] In some embodiments, the amino acid sequence comprises LGCDE (SEQ ID NO: 3), or functional variants thereof comprising about 60% or 80% sequence identity to SEQ ID NO: 3. In some embodiments, the disclosure relates to an amino acid sequence comprising an amino acid comprising at least 80% sequence identity to SEQ ID NO: 3. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 3 comprising about 4 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%. 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 3 from which it was derived.

[0103] In some embodiments, the amino acid sequence comprises MARRQESICGRPWTWTPGLLMLIILGIHQGSGQGQGSRLRQEDFPPRIVEHPSDVIV SKGEPTTLNCKAEGRPTPTIEWYKDGERVETDKDDPRSHRMLLPSGSLFFLRIVHGR RSKPDEGSYVCVARNYLGEAVSRNASLEVALLRDDFRQNPTDVVVAAGEPAILECQPPRGHPEPTIYWKKDKVRIDDKEERISIRGGKLMISNTRKSDAGMYTCVGTNMVGE RDSDPAELTVFERPTFLRRPINQVVLEEEAVEFRCQVQGDPQPTVRWKKDDADLPR GRYDIKDDYTLRIKKAMSTDEGTYVCIAENRVGKVEASATLTVRVRPVAPPQFVVR PRDQIVAQGRTVTFPCETKGNPQPAVFWQKEGSQNLLFPNQPQQPNSRCSVSPTGDL TITNIQRSDAGYYICQALTVAGSILAKAQLEVTDVLTDRPPPIILQGPINQTLAVDGTA LLKCKATGEPLPVISWLKEGFTFLGRDPRATIQDQGTLQIKNLRISDTGTYTCVATSS SGETSWSAVLDVTESGATISKNYDMNDLPGPPSKPQVTDVSKNSVTLSWQPGTPGV LPASAYIIEAFSQSVSNSWQTVANHVKTTLYTVRGLRPNTIYLFMVRAINPQGLSDPS PMSDPVRTQDISPPAQGVDHRQVQKELGDVVVRLHNPVVLTPTTVQVTWTVDRQP QFIQGYRVMYRQTSGLQASTVWQNLDAKVPTERSAVLVNLKKGVTYEIKVRPYFN EFQGMDSESKTVRTTEEAPSAPPQSVTVLTVGSHNSTSISVSWDPPPADHQNGIIQEY KIWCLGNETRFHINKTVDAAIRSVVIGGLFPGIQYRVEVAASTSAGVGVKSEPQPIII GGRNEVVITENNNSITEQITDVVKQPAFIAGIGGACWVILMGFSIWLYWRRKKRKG LSNYAVTFQRGDGGLMSNGSRPGLLNAGDPNYPWLADSWPATSLPVNNSNSGPNEI GNFGRGDVLPPVPGQGDKTATMLSDGAIYSSIDFTTKTTYNSSSQITQATPYATTQIL HSNSIHELAVDLPDPQWKSSVQQKTDLMGFGYSLPDQNKGNNALLYIPDYRLAEG LSNRMPHNQSQDFSTTSSHNSSERSGSLSGGKGGKKKKTKNSSKAQKNNGSTWAN VPLPPPPVQPLPGTELGHYAAEQENGYDSDSWCPPLPVQTYLHQGMEDELEEDEDR VPTPPVRGVASSPAISFGQQSTATLTPSPREEMQPMLQAHLDELTRAYQFDIAKQTW HIQSNTPPPQPPAPPLGYVSGALISDLETDVPDEDADDEEEPLEIPRPLRALDQTPGSS MDNLDSSVTGKAFSSSQRQRPTSPFSTDSNTSAAQNQSQRPRPTKKHKGGRMDPQP VLPHRREGMPDDLPPPPDPPPGQGLRQQIGLSQHSGNVENSTERKGSSLERQQAAN LEDTKSSLDCPAKTVLEWQRQTQDWINSTERQEETRKAPHKQGVGSEESLVPYSKP SFPSPGGHSSSGTSSSKGSTGPRKADVLRGSHQRNANDLLDIGYVGSNSQGQFTE (SEQ ID NO: 4), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 4. In some embodiments, the disclosure relates to an amino acid sequence comprising at least about 75% sequence identity to SEQ ID NO: 4. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 4 comprising about 4. about 5. about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids,about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, or about 1521 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, or about 1500 amino acids to and including a higher number of amino acids selected from one of about 8. about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, or about 1521 amino acids. In some embodiments, the functional fragment has at least about 75%. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%. 97%. 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 4 from which it was derived.

[0104] In some embodiments, the amino acid sequence comprises PRIVEHPSDVIVSKGEPTTLNCKAEGRPTPTIEWYKDGERVETDKDDPRSHRMLLPS GSLFFLRIVHGRRSKPDEGSYVCVARNYLGEAVSRNASLEVA (SEQ ID NO: 5). or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 5. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 5. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 5 comprising about 4, about 5. about 6. about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 98 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30. about 40, about 50, about 60. about 70. about 80, or about 90 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 98 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 5 from which it was derived.

[0105] In some embodiments, the amino acid sequence comprisesNGIIQEYKIWCLGNETRFHINKTVDAAIRSVVIGGLFPGIQYRVEVAASTSAGVGVK SEPQPIIIGGRNEVVITENNNSITEQITDVVKQPAFIAGIGGACWVILMGFSIWLYWRR KKRKGLSNYAVTFQRGDGGLMSNGSRPGLLNAGDPNYPWLADSWPATSLPVNNSN SGPNEIGNFGRGDVLPPVPGQGDKTATMLSDGAIYSSIDFTTKTTYNSSSQITQATPY ATTQILHSNSIHELAVDLPDPQWKSSVQQKTDLMGFGYSLPDQNKGNNALLYIPD (SEQ ID NO: 6), or functional variants thereof comprising about 75%. 80%. 85%. 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 6. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity7to SEQ ID NO: 6. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 6 comprising about 4, about 5. about 6, about 7, about 8. about 9, about 10, about 20. about 30. about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 250, or about 283 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8. about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, or about 250 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20. about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 250, or about 283 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity7to the amino acid sequence within SEQ ID NO: 6 from which it was derived.

[0106] In some embodiments, the amino acid sequence comprises VVVRLHNPVVLTPTTVQVTWTVDRQPQFIQGYRVMYRQTSGLQASTVWQNLDAK VPTERSAVLVNLKKGVTYEIKVRPYFNEFQGMDSESKTVRTTEEA (SEQ ID NO: 7), or functional variants thereof comprising about 75%, 80%, 85%. 90%. 91%. 92%. 93%. 94%, 95%, 96%, 97%, 98% or about 99% sequence identity7to SEQ ID NO: 7. In someembodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 7. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 7 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 98 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20. about 30. about 40, about 50, about 60. about 70. about 80, or about 90 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 98 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%. 90%. 91%. 92%. 93%. 94%. 95%. 96%. 97%. 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 7 from which it was derived.

[0107] In some embodiments, the amino acid sequence comprises AFIAGIGGACWVILMGFSIWLYW (SEQ ID NO: 8), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%. 93%. 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 8. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 8. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 8 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, or about 22 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, or about 20 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, or about 22 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%. 94%. 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 8 from which it was derived.

[0108] In some embodiments, the amino acid sequence comprises RRKKRKGLSNYAVTFQRGDGGLMSNGSRPGLLNAGDPNYPWLADSWPATSLPVN NSNSGPNEIGNFGRGDVLPPVPGQGDKTATMLSDGAIYSSIDFTTKTTYNSSSQITQA TPYATTQILHSNSIHELAVDLPDPQWKSSVQQKTDLMGFGYSLPDQNKGNN ALLYIP DYRLAEGLSNRMPHNQSQDFSTTSSHNSSERSGSLSGGKGGKKKKTKNSSKAQKN NGSTWANVPLPPPPVQPLPGTELGHYAAEQENGYDSDSWCPPLPVQTYLHQGMED ELEEDEDRVPTPPVRGVASSPAISFGQQSTATLTPSPREEMQPMLQAHLDELTRAYQF DIAKQTWHIQSNTPPPQPPAPPLGYVSGALISDLETDVPDEDADDEEEPLEIPRPLRALDQTPGSSMDNLDSSVTGKAFSSSQRQRPTSPFSTDSNTSAAQNQSQRPRPTKKHK GGRMDPQPVLPHRREGMPDDLPPPPDPPPGQGLRQQIGLSQHSGNVENSTERKGSS LERQQAANLEDTKSSLDCPAKTVLEWQRQTQDWINSTERQEETRKAPHKQGVGSE ESLVPYSKPSFPSPGGHS S S GTS S SKGSTGPRKADVLRGSHQRNANDLLDIGYVGSN SQGQFTE (SEQ ID NO: 9), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%. 94%. 95%. 96%. 97%. 98% or about 99% sequence identity to SEQ ID NO: 9. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 9. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 9 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60. about 70, about 80, about 90. about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, or about 625 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5, about 6. about 7, about 8, about 9. about 10. about 20. about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, or about 600 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, or about 625 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 9 from which it was derived.

[0109] In some embodiments. the amino acid sequence comprises MSGIGWQTLSLSLGLVLSILNKVAPQACPAQCSCSGSTVDCHGLALRSVPRNIPRNT ERLDLNGNNITRITKIDFAGLRHLRVLQLMENRISTIERGAFQDLKELERLRLNRNNL QLFPELLFLGTAKLYRLDLSENQIQAIPRKAFRGAVDIKNLQLDYNQISCIEDGAFRA LRDLEVLTLNNNNITRLSVASFNHMPKLRTFRLHSNNLYCDCHLAWLSDWLRQRPR VGLYTQCMGPSHLRGHNVAEVQKREFVCSDEEEGHQSFMAPSCSVLHCPAACTCSNNIVDCRGKGLTEIPTNLPETITEIRLEQNSIRVIPPGAFSPYKKLRRLDLSNNQISELA PDAFQGLRSLNSLVLYGNKITELPKSLFEGLFSLQLLLLNANKINCLRVDAFQDLHN LNLLSLYDNKLQTVAKGTFSALRAIQTMHLAQNPFICDCHLKWLADYLHTNPIETS GARCTSPRRLANKRIGQIKSKKFRCSAKEQYFIPGTEDYRSKLSGDCFADLACPEKC RCEGTTVDCSNQRLNKIPDHIPQYTAELRLNNNEFTVLEATGIFKKLPQLRKINFSNN KITDIEEGAFEGASGVNEILLTSNRLENVQHKMFKGLESLKTLMLRSNRISCVGNDS FIGLGSVRLLSLYDNQITTVAPGAFDSLHSLSTLNLLANPFNCNCHLAWLGEWLRRK RIVTGNPRCQKPYFLKEIPIQDVAIQDFTCDDGNDDNSCSPLSRCPSECTCLDTVVRC SNKGLKVLPKGIPKDVTELYLDGNQFTLVPKELSNYKHLTLIDLSNNRISTLSNQSFS NMTQLLTLILSYNRLRCIPPRTFDGLKSLRLLSLHGNDISVVPEGAFNDLSALSHLAI GANPLYCDCNMQWLSDWVKSEYKEPGIARCAGPGEMADKLLLTTPSKKFTCQGP VDITIQAKCNPCLSNPCKNDGTCNNDPVDFYRCTCPYGFKGQDCDVPIHACISNPC KHGGTCHLKEGENAGFWCTCADGFEGENCEVNIDDCEDNDCENNSTCVDGINNY TCLCPPEYTAANLNEVEKGELCEEKLDFCAQDLNPCQHDSKCILTPKGFKCDCTPG YIGEHCDIDFDDCQDNKCKNGAHCTDAVNGYTCVCPEGYSGLFCEFSPPMVLPRTS PCDNFDCQNGAQCIIRINEPICQCLPGYLGEKCEKLVSVNFVNKESYLQIPSAKVRPQ TNITLQIATDEDSGILLYKGDKDHIAVELYRGRVRASYDTGSHPASAIYSVETINDGN FHIVELLTLDSSLSLSVDGGSPKVITNLSKQSTLNFDSPLYVGGMPGKNNVASLRQA PGQNGTSFHGCIRNLYINSELQDFRKMPMQTGILPGCEPCHKKVCAHGMCQPSSQS GFTCECEEGWMGPLCDQRTNDPCLGNKCVHGTCLPINAFSYSCKCLEGHGGVLCD EEEDLFNPCQMIKCKHGKCRLSGVGQPYCECNSGFTGDSCDREISCRGERTRDYYQ KQQGYAACQTTKKVSRLECRGGCAGGQCCGPLRSKRRKYSFECTDGSSFVDEVEK VVKCGCARCAS (SEQ ID NO: 10), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 10. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 10. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 10 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40. about 50. about 60, about 70, about 80, about 90, about 100, about 110. about 120. about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500amino acids, or about 1541 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, or about 1500 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80. about 90, about 100. about 110, about 120, about 130. about 140. about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 ammo acids, or about 1541 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity7to the amino acid sequence within SEQ ID NO: 10 from which it was derived.

[0110] In some embodiments. the amino acid sequence comprises SVLHCPAACTCSNNIVDCRGKGLTEIPTNLPETITEIRLEQNSIRVIPPGAFSPYKKLRR LDLSNNQISELAPDAFQGLRSLNSLVLYGNKITELPKSLFEGLFSLQLLLLNANKINC LRVDAFQDLHNLNLLSLYDNKLQTVAKGTFSALRAIQTMHLAQNPFICDCHLKWL ADYLHTNPIETSGARCTSPRRLANKRIGQIKSKKFRCSAKEQYFI (SEQ ID NO: 11), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 11. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 11. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 11 comprising about 4, about 5. about 6. about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, or about 217 amino acids, or anumber of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, or about 200 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, or about 218 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%. 90%. 91%. 92%. 93%. 94%. 95%. 96%. 97%. 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 11 from which it was derived.[OHl] In some embodiments, the amino acid sequence comprisesSRCPSECTCLDTVVRCSNKGLKVLPKGIPKDVTELYLDGNQFTLVPKELSNYKHLTL IDLSNNRISTLSNQSFSNMTQLLTLILSYNRLRCIPPRTFDGLKSLRLLSLHGNDISVV PEGAFNDLSALSHLAIGANPLYCDCNMQWLSDWVKSEYKEPGIARCAGPGEMAD KLLLTTPSKKFTCQGP (SEQ ID NO: 12), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 12. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 12. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 12 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40. about 50, about 60, about 70, about 80, about 90, about 100, about 110. about 120, about 130, about 140, about 150. about 160, about 170, about 180, or about 185 amino acids, or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80. about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, or about 180 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, or about 185 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%. 94%. 95%. 96%. 97%. 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 12 from which it was derived.

[0112] In some embodiments, the disclosure relates to a compound that inhibits the activity of an amino acid sequence and compositions or pharmaceutical compositions comprising the compound. In some embodiments, the disclosure relates to a plurality of compounds that inhibit the activity of the amino acid sequence and compositions orpharmaceutical compositions comprising the plurality of compounds. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the compound or the plurality of compounds. In some embodiments, the amino acid sequence is one from Table 2 or a functional variant thereof.

[0113] In some embodiments, the disclosure relates to a compound or a plurality of compounds that inhibits expression of a nucleic acid sequence encoding the amino acid sequence and compositions or pharmaceutical composition comprising the compound or plurality of compounds. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the compound or plurality of compounds. In some embodiments, the nucleic acid sequence encodes an amino acid sequence of Table 2 or functional variants thereof.Table 2

[0114] In some embodiments, the compound or plurality of compounds that inhibits an amino acid sequence comprises at least one EP300 modulator. In some embodiments, the at least one EP 300 modulator comprises at least one or more amino acid sequences herein, EP300i, an analog of EP300i, Y08197 an analog of Y08197, CCS1477, an analog of CCS1477, CPI-1612, an analog of CPI-1612, DS17701585, an analog of DS17701585, DS- 9300, an analog of DS-9300, NEO3734, an analog of NEO3734, a derivative of any of the foregoing, and a pharmaceutically acceptable salt one or more of the foregoing.

[0115] In some embodiments, the disclosed compositions or plurality of compounds inhibits E300 pathway activity by targeting an amino acid sequence comprising MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTN GGDISQLQTSLGIVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQAMASQAQQNS PGLSLINSMVKSPMAQTGLTSPNMGIGSSGPNQGPTQSPAGMMNSPVNQPAMGMN TGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRPNMQYPNAGMGNAGSL LTEPLQQGSPQMGGQPGLRGPQPLKMGMMNNPSPYGSPYTQNSGQQIGASGLGLQ IQTKTVLPNNLSPFAMDKKAVPGGGMPSMGQQPTPSVQQPGLVTPVAAGMGSGAH TADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHC QSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQSILTGAPVGLG NPSSLGVGQQSTPSLSTVSQIDPSSIERAYAALGLPYQVNQIPPQPQVQAKNQQSQPS GQSPQGMRSVNNMSASPMGVNGGVGVQTPNLLSDSMLHSTINSQNPMMSENAGV ASLGPLPTAAQPSSTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMEN LVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAP GMGPVPMNTGSNMGQQPTGMTTNGPVPDPSMIRGSVPNHMMPRMTPQPGLNQFG QMNMPQPPIGPRQPSPLQHHGQLAQSGSLNPPMGYGPRMQQASGQNQFLSQTQFT SQGMNVTNMPLAPSSGQAPVSQAQMSSSSCPVNSPIMPPGSQGSHIHCPTLPQQAH QNSPSPVPSRTPTPHHTPPSIGNQPPPATAIPTPVPTPPAIPPGPQPPSLHPSSRQTPTPPT HLPPQVQPSLPAAPSADQSQQQPRSQQSTAVSVPTPTAPLLPPQPSTPLSQPAVSIEGQ VSNPPSTSSTEVNSQTIPEKQPSQEVKMESKMEVDKPEPADAQPEDTKEAKGEDVK VEPTEMEERGPELKTDGKEEEEQPSTSATQSSPAPGQSKKKIFKPEELRQALMPTLE ALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRKLDTGQYQEPWQYIDD IWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSLGYCCGRKLEFSPQTLC CYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQGESVSLGDDPSQPQTTINKEQFSK RKNDTLDPELFVECTECGRKMHQICVLHHEIIWPSGFVCDGCLKKTARTRKENKLS AKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGMKARFV DSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDS VHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQKIP KPKRLQEWYKKMLDKAVSERIVHDYKDILKQATEDRLTSAKELPYFEGDFWPNVL EESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKK KPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIACPAPNSLPPIVDPDPLIPCDLMDG RDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQDRFVYTCNECKHHVETR WHCTVCEDYDLCITCYNTKNHDHKMEKLGLGLDDESNNQQAAATQSPGDSRRLSI QRCIQSLVHACQCRNANCSLPSCQKMKRVVQHTKGCKRKTNGGCPICKQLIALCCYHAKHCQENKCPVPFCLNIKQKLRQQQLQHRLQQAQMLRRRMASMQRTGVAGQ QQGLPSPTPATPTTPTGQQPATPQTPQPQPTSQPQPTPPNNMTPYLPRTQTTGPVSQG KAPGQVTPPTPPQTAQAPLPGPPPAAVEMAMQIQRAAETQRQMAHVQIFQRPIQHQ MPQMSPMAPMGMNPPPMARGPGGHLDPGIGPAGMQQQPPWAQGGMPQPQQMQS GMPRPAMMSVAQHGQPLNMAPQPGLGQVGVSPLKPGTVSQQALQNLLRTLRSPSS PLQQQQVLSILHANPQLLAAFIKQRAAKYANPNPQPLPGQPGMTQGQPGLQPPTMP GQQGVHSNPALQNMNPLQAGVQRAGLPQQQPQQQLQPPMGAMSPQAQQMNMN HNTMPSQFRDILRRQMMQQQGAGPGIGPGMANQFQQPQGIGYPPQQQQQQRMQH HMQQMQQGNMGQMGQLPQALGAEAGASLQAYQQRLLQQQMGSPAQPNPMSPQ QHMLPNQAQSPHLQGQQIPNSLSNQVRSPQPVPSPRPQSQPPHSSPSPRMQPQPSPH HVSPQTSSPHPGLVAAQAANPMEQGHFASPDQNSMLSQLASNPGMANLHGASATD LGLSSDNADLNSNLSQSTLDIH (SEQ ID NO: 13), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 13. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 13. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 13 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150. about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, about 2000 amino acids, or about 2411 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, or about 2000 amino acids to and including a higher number of amino acids selected from one of about 8. about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100,about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, about 2000 amino acids, or about 2411 amino acids. In some embodiments, the functional fragment has at least about 75%. 80%. 85%. 90%. 91%. 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 13 from which it was derived.

[0116] In some embodiments, the disclosed compositions or plurality of compounds inhibits E300 pathway activity by targeting an amino acid sequence comprising QQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAH CASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQSILTGAPVGLGN (SEQ ID NO: 14), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%. 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 14. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 14. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 14 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30. about 40, about 50, about 60, about 70. about 80. about 90, or about 99 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, or about 90 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20. about 30, about 40, about 50, about 60, about 70, about 80. about 90, or about 99 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 14 from which it was derived.

[0117] In some embodiments, the disclosed compositions or plurality of compounds inhibits E300 pathway activity by targeting an amino acid sequence comprising (SEQ ID NO: 15), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 15. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 15. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 15 comprising about 4, about 5, about 6,about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80. about 90, or about 99 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, or about 90 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20. about 30, about 40, about 50, about 60, about 70, about 80. about 90, or about 99 amino acids. In some embodiments, the functional fragment has at least about 75%. 80%. 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 15 from which it was derived.

[0118] In some embodiments, the disclosed compositions or plurality of compounds inhibits E300 pathway activity by targeting an amino acid sequence comprising GIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYARKVEGDMY ESANNRAEYYHLLAEKIYKIQKELE (SEQ ID NO: 16), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 16. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 16. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 16 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60. about 70, or about 79 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6. about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, or about 70 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60. about 70, or about 79 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 16 from which it was derived.

[0119] In some embodiments, the disclosed compositions or plurality7of compounds inhibits E300 pathway activity by targeting an amino acid sequence comprising IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRK LDTGQYQEPWQYIDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSL GYCCGRKL (SEQ ID NO: 17), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%. 96%. 97%. 98% or about 99% sequence identity7to SEQ ID NO: 17. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 17. In some embodiments, theamino acid sequence comprises a functional fragment of SEQ ID NO: 17 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10. about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, or about 120 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, or about 110 amino acids to and including a higher number of amino acids selected from one of about 8. about 9, about 10, about 20, about 30, about 40. about 50, about 60, about 70, about 80, about 90, about 100, about 110, or about 120 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 17 from which it was derived.

[0120] In some embodiments, the disclosed compositions or plurality of compounds inhibits E300 pathway activity by targeting an amino acid sequence comprising ENKLSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGM KARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYI SYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPP DQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDILKQATEDRLTSAKELPYFEGDF WPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIACPAPNSLPPIVDPDPLIPCD LMDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD (SEQ ID NO: 18), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 18. In some embodiments, the disclosure relates to an amino acid sequence comprising at least 75% sequence identity to SEQ ID NO: 18. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 18 comprising about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, or about 379 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90. about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, or about 300 amino acids to and including a higher number of amino acidsselected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, or about 379 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 18 from which it was derived.

[0121] In some embodiments, the nucleic acid sequence that the compound or plurality of compounds inhibits the expression of an amino acid sequence comprising MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDISQLQTSLGIVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQAMASQAQQNS PGLSLINSMVKSPMAQTGLTSPNMGIGSSGPNQGPTQSPAGMMNSPVNQPAMGMN TGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRPNMQYPNAGMGNAGSL LTEPLQQGSPQMGGQPGLRGPQPLKMGMMNNPSPYGSPYTQNSGQQIGASGLGLQ IQTKTVLPNNLSPFAMDKKAVPGGGMPSMGQQPTPSVQQPGLVTPVAAGMGSGAH TADPEKRKLIQQQLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHC QSGKSCQVAHCASSRQIISHWKNCTRHDCPVCLPLKNAGDKRNQQSILTGAPVGLG NPSSLGVGQQSTPSLSTVSQIDPSSIERAYAALGLPYQVNQIPPQPQVQAKNQQSQPS GQSPQGMRSVNNMSASPMGVNGGVGVQTPNLLSDSMLHSTINSQNPMMSENAGV ASLGPLPTAAQPSSTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMEN LVAYARKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAP GMGPVPMNTGSNMGQQPTGMTTNGPVPDPSMIRGSVPNHMMPRMTPQPGLNQFG QMNMPQPPIGPRQPSPLQHHGQLAQSGSLNPPMGYGPRMQQASGQNQFLSQTQFTSQGMNVTNMPLAPSSGQAPVSQAQMSSSSCPVNSPIMPPGSQGSHIHCPTLPQQAH QNSPSPVPSRTPTPHHTPPSIGNQPPPATAIPTPVPTPPAIPPGPQPPSLHPSSRQTPTPPT HLPPQVQPSLPAAPSADQSQQQPRSQQSTAVSVPTPTAPLLPPQPSTPLSQPAVSIEGQ VSNPPSTSSTEVNSQTIPEKQPSQEVKMESKMEVDKPEPADAQPEDTKEAKGEDVK VEPTEMEERGPELKTDGKEEEEQPSTSATQSSPAPGQSKKKIFKPEELRQALMPTLE ALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRKLDTGQYQEPWQYIDD IWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSLGYCCGRKLEFSPQTLC CYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQGESVSLGDDPSQPQTTINKEQFSK RKNDTLDPELFVECTECGRKMHQICVLHHEIIWPSGFVCDGCLKKTARTRKENKLS AKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHASDKTVEVKPGMKARFV DSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQKIP KPKRLQEWYKKMLDKAVSERIVHDYKDILKQATEDRLTSAKELPYFEGDFWPNVL EESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKK KPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIACPAPNSLPPIVDPDPLIPCDLMDG RDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQDRFVYTCNECKHHVETR WHCTVCEDYDLCITCYNTKNHDHKMEKLGLGLDDESNNQQAAATQSPGDSRRLSI QRCIQSLVHACQCRNANCSLPSCQKMKRVVQHTKGCKRKTNGGCPICKQLIALCC YHAKHCQENKCPVPFCLNIKQKLRQQQLQHRLQQAQMLRRRMASMQRTGVAGQ QQGLPSPTPATPTTPTGQQPATPQTPQPQPTSQPQPTPPNNMTPYLPRTQTTGPVSQG KAPGQVTPPTPPQTAQAPLPGPPPAAVEMAMQIQRAAETQRQMAHVQIFQRPIQHQ MPQMSPMAPMGMNPPPMARGPGGHLDPGIGPAGMQQQPPWAQGGMPQPQQMQS GMPRPAMMSVAQHGQPLNMAPQPGLGQVGVSPLKPGTVSQQALQNLLRTLRSPSS PLQQQQVLSILHANPQLLAAFIKQRAAKYANPNPQPLPGQPGMTQGQPGLQPPTMP GQQGVHSNPALQNMNPLQAGVQRAGLPQQQPQQQLQPPMGAMSPQAQQMNMN HNTMPSQFRDILRRQMMQQQGAGPGIGPGMANQFQQPQGIGYPPQQQQQQRMQH HMQQMQQGNMGQMGQLPQALGAEAGASLQAYQQRLLQQQMGSPAQPNPMSPQ QHMLPNQAQSPHLQGQQIPNSLSNQVRSPQPVPSPRPQSQPPHSSPSPRMQPQPSPH HVSPQTSSPHPGLVAAQAANPMEQGHFASPDQNSMLSQLASNPGMANLHGASATD LGLSSDNADLNSNLSQSTLDIH (SEQ ID NO: 13), or functional variants thereof comprising about 75%, 80%, 85%, 90%, 91%. 92%. 93%. 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to SEQ ID NO: 13. In some embodiments, the amino acid sequence comprises at least 75% sequence identity to SEQ ID NO: 13. In some embodiments, the amino acid sequence comprises a functional fragment of SEQ ID NO: 13 comprising about 4, about 5. about 6, about 7. about 8, about 9. about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, about 2000 amino acids, or about 2411 amino acids or a number of amino acids in a range from and including any one of about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 20. about 30, about 40, about 50. about 60, about 70, about 80, about 90. about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, or about 2000 amino acids to and including a higher number of amino acids selected from one of about 8, about 9, about 10, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160 amino acids, about 170 amino acids, about 180 amino acids, about 190 amino acids, about 200 amino acids, about 300 amino acids, about 400 amino acids, about 500 amino acids, about 600 amino acids, about 700 amino acids, about 800 amino acids, about 900 amino acids, about 1000 amino acids, about 1100 amino acids, about 1200 amino acids, about 1300 amino acids, about 1400 amino acids, about 1500 amino acids, about 2000 amino acids, or about 2411 amino acids. In some embodiments, the functional fragment has at least about 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or about 99% sequence identity to the amino acid sequence within SEQ ID NO: 13 from which it was derived.

[0122] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from EP300i, an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. EP300i has the structure of formula I:Formula I.

[0123] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from Y08197, an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. Y08197 is 3-Acetyl-N-(2-fluoro-5-(hydroxymethyl)-3-(l-methyl-lH- pyrazol-4-yl)phenyl)-7-methoxyindolizine-l -carboxamide, and has the structure of formula II:

[0124] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from CCS 1477, an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. CCS1477 has the structure of formula III:Formula III

[0125] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from CPI- 1612, an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. CPI-1612 has the structure of formula IV:Formula IV.

[0126] In some embodiments, a CPI-1612 analog includes is selected from compounds disclosed in Wilson, J.E. et al. “Discovery of CPI-1612: A Potent, Selective, and Orally Bioavailable EP300 / CBP Histone Acetyltransferase Inhibitor,” (2020) ACS Med. Chem. Lett. 11(6): 1324-1329, which is incorporated herein by reference as if fully set forth.

[0127] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from DS 17701585, an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. DS17701585 has the structure of formula V:Formula V.

[0128] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from DS 17701585, an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. DS17701585 has the structure of formula VI:Formula VI.

[0129] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from DS- 9300 , an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. DS-9300 has the structure of formula VIEFormula VII.

[0130] In some embodiments, the EP300 modulator is an EP300 inhibitor chosen from NEO3734, an analog thereof, a derivative of the foregoing, or a pharmaceutically acceptable salt of the foregoing. NEO3734 has the structure of formula VIII:Formula VIII.

[0131] Methods of preparing and administering EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure to a subject in need thereof are well known to or are readily determined by those skilled in the art. The invention relates to the manufacturing of a synthetic polypeptide which is an amino acid sequence or fragment thereof that acts as a hormone or dual tropic hormone involved in glucose metabolism. In the synthetic polypeptide, from about 14% to about 50% of the a-amino acid residues found in the biologically active polypeptide or fragment are replaced with amino acid residues. In another embodiment of the invention, the a-amino acid residues and the amino acid residues are distributed in a repeating pattern, the analog is then covalently or noncovalently bound to a pharmaceutical agent to increase its stability and alter its pharmacokinetic profile. Human cells are then contacted with the pharmaceutical agent ligated to the molecule to induce the biochemical pathway or biological activity ordinarily induced by the naturally occurring polypeptide upon which the analog is based.

[0132] The compositions of the disclosure may be prepared by the synthetic chemical procedures described herein, as well as other procedures similar to those which may be used for making p-amino acid peptides. Such procedures include both solution and solid phase procedures, e.g., using either Boc and Fmoc methodologies. The compounds of the invention may be synthesized using solid phase synthesis techniques. Fmoc-N-Protected amino acids can be used to synthesize polypeptides by conventional manual solid-phase synthesis procedures under standard conditions on any number of solid supports, including ortho-chloro- trityl chloride resin. Esterification of Fmoc-amino acids with the ortho-chloro-trityl resin can be performed according to the method of Barlos et. al., Tetrahedron Lett., 1989, 30, 3943. The resin (150 mg, 1.05 mmol Cl) is swelled in 2 ml CH2CI2 for 10 min. A solution of the Fmoc- protected P-amino acid in CH2CI2 and iPrcEtN are then added successively and the suspension is mixed under argon for 4 h. Subsequently, the resin is filtered and washed with CH2C12 / MeOH / iPr2EtN (17:2: 1, 3x3 min), CH2CI2 (3x3 mm), DMF (2x3 mm), CH2CI2 (3x3 min), and MeOH (2x3 min). The substitution of the resin is determined on a 3 mg sample by measuring the absorbance of the dibenzofulvene adduct at 300 nm. The Fmoc group is removed using 20% piperidine in DMF (4 ml, 2x20 min) under Ar bubbling. The resin is then filtered and washed with DMF (6x3 min). For each coupling step, a solution of the amino acid (3 equiv.). BOP (3 equiv.) and HOBT (3 equiv.) in DMF (2 ml) and iPnEtN (9 eq) are added successively to the resin and the suspension is mixed for 1 h under Ar. Monitoring of the coupling reaction is performed with 2,4,6-trinitrobenzene-sulfonic acid (TNBS) (W.S. Hancock and J.E. Battersby, Anal. Biochem. (1976), 71, 260). In the case of a positive TNBS test (indicating incomplete coupling), the suspension is allowed to react for a further 1 h. The resin is then filtered and washed with DMF (3x3 min) prior to the following Fmoc deprotection step. After the removal of the last Fmoc protecting group, the resin is washed with DMF (6x3 min), CH2CI2 (3x3 min), Et2O (3x3 min) and dried under vacuum for 3 h. Finally the peptides are cleaved from the resin using 2% TFA in CH2CI2 (2 ml, 5x15 min) under Ar. The solvent is removed and the oily residues are triturated in ether to give the crude polypeptides. The compounds are further purified by high performance liquid chromatography (HPLC).

[0133] The compositions of the invention may be prepared by the synthetic chemical procedures described herein, as well as other procedures similar to those which may be used for making amino acid containing peptide sequences. Such procedures include both solution and solid phase procedures, e.g.. using either Boc or Fmoc methodologies. The compounds of the invention may be synthesized using solid phase synthesis techniques. Fmoc-N-Protectedp-amino acids can be used to synthesize polypeptides by conventional manual solid-phase synthesis procedures under standard conditions on any number of solid supports, including ortho-chloro-trityl chloride resin, Wang resin (NovaBiochem 75mmol substitution) and Rink amid resin (NovaBiochem ,55mmol substitution). Resin is typically swelled in 100% DMF for 30 minutes then deprotected using 20% piperidine in DMF for 2 minutes at 80° (3x). Fmoc protected amino acids (natural or non-natural) can then be coupled to the resin using a cocktail of AA:HATU:DIEA:Resin (3:2.5:4: 1, LiCL 0.8M final concentration) in DMF for 2 minutes at 70° (3x). The resin is then washed (3x) with DMF, DCM (dichloromethane) (3x) and again with DMF (3x) between deprotection and coupling steps. Monitoring of the coupling reaction is performed with 2.4.6-trinitrobenzene-sulfonic acid (TNBS) (W.S. Hancock and J.E. Battersby, Anal. Biochem. (1976), 71, 260). In the case of a positive TNBS test (indicating incomplete coupling), the suspension is allowed to react for another three times. This process is repeated until the desired product has been achieved. After the removal of the last Fmoc protecting group, the resin is washed with DMF (3x), CH2CI2 (3x) and DMF again (3x). The remaining free-amine group is then acetylated using a cocktail of DIEA:Ac2O (1: 1) for 5 minutes at room temperature. Full-length peptides were then cleaved from solid support using TFA:TIS:H2O (95:2.5:2.5) for 150 minutes, precipitated in cold ethyl ether and lyophilized. The polymer was reconstituted in a 1 : 1 solution of A:B (A: H2O, 0.1% TFA) (B: 90: 10:0.1 acetonitrile / H2O / TFA).

[0134] The compositions described herein may be prepared by successive amide bondforming procedures in which amide bonds are formed between the amino group of a first

[0135] amino acid residue or a precursor thereof and the a-carboxyl group of a second amino acid residue or amino acid residue or a precursor thereof. The amide bond-forming step may be repeated as many times, and with specific amino acid residues and / or precursors thereof, as required to give the desired polypeptide. Also analogs comprising two, three, or more amino acid residues may be joined together to yield larger analogs comprising any combination of amino acids. Cyclic compounds may be prepared by forming peptide bonds between the N-terminal and C-terminal ends of a previously synthesized linear polypeptide or through the disulfide crosslinking of sidechains of non-adjacent residues. p3-amino acids may be produced enantioselectively from corresponding p-amino acids. For instance, by Amdt- Eisert homologation of N-protected a-amino acids. Homologation may be followed by coupling of the reactive diazoketone intermediate of the Wolff rearrangement with a p-amino acid residue.

[0136] The route of administration of the EP300 modulator, or antigen-binding fragment, variant, or derivative thereof may be. for example, oral, parenteral, by inhalation or topical. The term parenteral as used herein includes, e.g., intravenous, intraarterial, intraperitoneal, intramuscular, subcutaneous, rectal or vaginal administration. While all these forms of administration are clearly contemplated as being within the scope of the disclosure, a form for administration would be a solution for injection, in particular for intravenous or intraarterial injection or drip. Usually, a suitable pharmaceutical composition for injection may comprise a buffer (e.g., acetate, phosphate or citrate buffer), a surfactant (e g., polysorbate), optionally a stabilizer agent (e.g., human albumin), etc. However, in other methods compatible with the teachings herein, EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can be delivered directly to the site of a wound thereby increasing the exposure of the wounded tissue to the therapeutic agent.

[0137] In some embodiments, EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure are administered in an amount sufficient to treat or prevent fibrosis. Treatment of fibrosis can be measured and observed by methods that are known in the art without undue experimentation. In this regard, it will be appreciated that the disclosed therapies will be formulated so as to facilitate administration and promote stability of the amnio acids or molecules that are active agents.

[0138] In some embodiments, pharmaceutical compositions in accordance with the present disclosure comprise a pharmaceutically acceptable, non-toxic, sterile carrier such as physiological saline, non-toxic buffers, preservatives and the like. For the purposes of the instant application, “an effective amount” of an EP300 modulator, or functional fragment, variant, or derivative thereof, conjugated or unconjugated, shall be held to mean an amount sufficient to achieve effective binding to a target and to achieve a desired goal benefit, e.g., to ameliorate symptoms or to abrogate severity of a disease or disorder in an animal model of disease or to detect a substance or a cell or a particular physiological parameter.

[0139] The pharmaceutical compositions used in this disclosure may comprise pharmaceutically acceptable carriers, including, e.g., ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty7acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0140] Preparations for parenteral administration includes sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. In the subject disclosure, pharmaceutically acceptable carriers include, but are not limited to, from about 0.01 to about 0.1M, about 0.05M phosphate buffer or about 0.8% saline. Other common parenteral vehicles include sodium phosphate solutions, Ringer’s dextrose, dextrose and sodium chloride, lactated Ringer’s, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers, such as those based on Ringer’s dextrose, and the like. Preservatives and other additives may also be present such as for example, antimicrobials, antioxidants, chelating agents, and inert gases and the like.

[0141] More particularly, pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and stenle powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In such cases, the composition must be sterile and should be fluid to the extent that easy syringability7exists. It should be stable under the conditions of manufacture and storage and will preferably be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

[0142] Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols, such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0143] In any case, sterile injectable solutions can be prepared by incorporating an active compound (e.g.. an EP300 modulator, or antigen-binding fragment, variant, or derivative thereof, by itself or in combination with other active agents) in the required amount in anappropriate solvent with one or a combination of ingredients enumerated herein, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle, which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, which yields a powder of an active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0144] Parenteral formulations may be a single bolus dose, an infusion or a loading bolus dose followed with a maintenance dose. These compositions may be administered at specific fixed or variable intervals, e.g., once a day, or on an “as needed” basis.

[0145] The amount of an EP300 modulator, or fragment, variant, or derivative thereof that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. The composition may be administered as a single dose, multiple doses or over an established period of time in an infusion. Dosage regimens also may be adjusted to provide the optimum desired response (e.g.. a therapeutic or prophylactic response in a non-human subject to mimic the response for a corresponding therapy if administered to a human subject).

[0146] In keeping with the scope of the present disclosure, EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure may be administered to a subject (e.g.. a non-human animal model of disease) in accordance with the aforementioned methods of administration in an amount sufficient to produce a desired effect. The EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can be administered to a subject in a conventional dosage form prepared by combining the antibody with a conventional pharmaceutically acceptable carrier or diluent according to known techniques. It will be recognized by one of skill in the art that the form and character of the pharmaceutically acceptable carrier or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other w ell-known variables. Those skilled in the art will further appreciate that a cocktail comprising one or more species of EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure may prove to be particularly effective or may be of particular interest for study in an animal model of disease.

[0147] Dosage regimens in the above methods of treatment and uses are adjusted to provide the optimum desired response (e.g., a therapeutic response). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation.Parenteral compositions may be formulated in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the present invention are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active compound for the treatment of wound healing or fibrosis.

[0148] The efficient dosages and the dosage regimens for the EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, depend on the disease or condition to be treated and may be determined by the persons skilled in the art. An exemplary, non-limiting range for a therapeutically effective amount of a compound of the present disclosure is from about 0.1 to about 100 mg / kg, such as from about 0.1 to about 50 mg / kg, for example from about 0.1 to about 20 mg / kg, such as from about 0.1 to about 10 mg / kg. for instance about 0.5,from about 0.3. about 1.0, or about 3 mg / kg.

[0149] A physician or veterinarian having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the EP300 modulator, or functional fragments, variants, or derivatives thereof, employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In general, a suitable daily dose of a composition of the present disclosure will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above. Administration may be intravenous, intramuscular, intraperitoneal, or subcutaneous, and for instance administered proximal to the site of the target. If desired, the effective daily dose of a pharmaceutical composition may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms.

[0150] In some embodiments, the EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, may be administered by infusion in a weekly dosage of from 10 to 500 mg / m2, such as of from 200 to 400 mg / m2. Such administration may be repeated, e.g., about 1 to 8 times, such as about 3 to 5 times. The administration may beperformed by continuous infusion over a period of from about 2 to about 24 hours, such as of from about 2 to about 12 hours.

[0151] In some embodiments, the EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, may be administered by slow continuous infusion over a long period, such as more than about 24 hours, in order to reduce toxic side effects.

[0152] In some embodiments, the EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, may be administered in a weekly dosage of from about 250 mg to about 2000 mg, such as for example about 300 mg, about 500 mg, about 700 mg, about 1000 mg, about 1500 mg or about 2000 mg, for up to about 8 times, such as from about 4 to about 6 times. The administration may be performed by continuous infusion over a period of from about 2 to about 24 hours, such as of from about 2 to about 12 hours. Such regimen may be repeated one or more times as necessary, for example, after about 6 months or about 12 months. The dosage may be determined or adjusted by measuring the amount of compound of the present invention in the blood upon administration by for instance taking out a biological sample and using anti-idiotypic antibodies which target the antigen binding region of the EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure.

[0153] In some embodiments, the EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, may be administered by maintenance therapy, such as, e.g., about once a week for a period of about 6 months or more.

[0154] In some embodiments, the EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, may be administered by a regimen including one infusion of an EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, followed by an infusion of an EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, conjugated to a radioisotope. The regimen may be repeated, e.g., about 7 to about 9 days later.

[0155] As non-limiting examples, treatment according to the present disclosure may be provided as a daily dosage of an EP300 modulator, or functional fragments, variants, or derivatives thereof in an amount of from about 0. 1 to about 100 mg / kg, such as about 0.5, 0.9, 1.0, 1.1, 1.5, 2. 3, 4. 5. 6, 7, 8. 9, 10, 11, 12, 13, 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26, 27, 28, 29, 30, 40, 45, 50, 60, 70, 80, 90 or 100 mg / kg, per day, on at least one of day 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40, or alternatively, at least one of week 1, 2, 3. 4, 5, 6, 7, 8. 9, 10, 11, 12, 13. 14. 15, 16, 17, 18, 19 or 20 after initiation of treatment, or anycombination thereof, using single or divided doses of every 24, 12, 8, 6, 4, or 2 hours, or any combination thereof.

[0156] In some embodiments, the dosage of EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, being administered may be from about 0. 1 mg to about 15,000 mg, In some embodiments, the dosage of EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, being administered may be from about 0.5 mg to about 12,000 mg. In some embodiments, the dosage of EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, being administered may be from about 1 mg to about 10,000 mg, In some embodiments, the dosage of EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, being administered may be from about 5 mg to about 5.000 mg, In some embodiments, the dosage of EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, being administered may be from about 5 mg to about 1,000 mg, In some embodiments, the dosage of EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure, being administered may be from about 10 mg to about 500 mg,

[0157] Effective doses of the compositions of the present disclosure vary depending upon many different factors, including means of administration, target site, physiological state of the subject, other medications administered, and whether what is the goal of the study in which the composition is being administered (e.g., testing a combination therapy for its effects in an animal disease model as a predictor of its efficacy or toxicity in a human subject, or testing the effect of healing in a wound healing model of a particular disease or disorder). For B-cell depletion, the dosage can range, e.g., from about 0.0001 to about 100 mg / kg, from about 0.01 to about 10 mg / kg (e.g., 0.02 mg / kg, 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1 mg / kg, 2 mg / kg, 5 mg / kg, 10 mg / kg, etc.), of the host body weight. For example, dosages can be about 1 mg / kg body weight or about 10 mg / kg body weight or within the range of about 1 mg / kg to about 10 mg / kg. In some embodiments, dosages are at least about 1 mg / kg. Doses intermediate in the above ranges are also intended to be within the scope of the disclosure. Subjects can be administered such doses daily, on alternative days, weekly or according to any other schedule determined by empirical analysis. Exemplary dosage schedules include about 1 to about 10 mg / kg or about 15 mg / kg on consecutive days, about 30 mg / kg on alternate days, or about 10 mg / kg or about 60 mg / kg weekly. In some methods, two or more monoclonal antibodies with different binding specificities are administered simultaneously, in which case the dosage of each antibody administered falls within the ranges indicated. In some embodiments, thecompositions of the present disclosure are administered in an amount of about 10 mg / kg every other week.

[0158] EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can be administered on multiple occasions. Intervals between single dosages can be weekly, monthly or yearly. Intervals can also be irregular as indicated by measuring blood levels of target polypeptide or target molecule in the subject. In some embodiments, dosage is adjusted to achieve a plasma polypeptide concentration of about 1 pg / ml to about 1000 pg / ml. and in some embodiments, about 1 pg / ml to about 30 pg / ml, or about 25 pg / ml to about 300 pg / ml. Alternatively, EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the subject. The half-life of an EP300 modulator can also be prolonged via fusion to a stable polypeptide or moiety, e.g., albumin or PEG. In some embodiments, EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can be administered in unconjugated form. In some embodiments, EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can be administered multiple times in conjugated form. In some embodiments, EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can be administered in unconjugated form, then in conjugated form, or vice versa.

[0159] The compositions of the present disclosure may be administered by any suitable method, e.g., intraparenterally, intraventricularly, intravenously, orally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrastemal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In some embodiments, the EP300 modulator, or functional fragments, variants, or derivatives thereof are administered in such a way that they cross the blood-brain barrier. This crossing can result from the physico-chemical properties inherent in the EP300 modulator molecule itself, from other components in a pharmaceutical formulation, or from the use of a mechanical device such as a needle, cannula or surgical instruments to breach the blood-brain barrier. Where the EP300 modulator is a molecule that does not inherently cross the blood-brain barrier, e.g., a fusion to a moiety that facilitates the crossing, suitable routes of administration are, e.g., directly into a wound on the face or body. In some embodiments, EP300 modulator are administered as a sustained release composition or device, such as a MEDIPAD™ device.

[0160] The compositions may also comprise an EP300 modulator dispersed in a biocompatible earner material that functions as a suitable delivery or support system for the compounds. Suitable examples of sustained release carriers include semipermeable polymer matrices in the form of shaped articles such as suppositories or capsules. Implantable or microcapsular sustained release matrices include polylactides, copolymers of L-glutamic acid and gamma-ethyl-L-glutamate; poly(2-hydroxyethyl-methacrylate), ethylene vinyl acetate, or poly-D-(-)-3hydroxybutyric acid.

[0161] EP300 modulator, or functional fragments, variants, or derivatives thereof of the disclosure can optionally be administered in combination with other agents e.g., to be tested for toxicity or for efficacy, e.g., in treating or having an effect on the disorder or condition in an animal model of disease. The agents can be administered simultaneously or in any order, or with a time interval in between.Cells

[0162] The disclosure relates to compositions of cells and, in some embodiments, compositions of one or a plurality of fibroblasts that comprise biologically active amounts of ROBO2 or functional fragments thereof. In some embodiments, the fibroblasts also comprise biologically active amounts of EID 1 or functional fragments thereof. In some embodiments, the cells further comprise Cdh4+. Nccunl+, and Gpc3 that encode CDH4, NCAM1 and GPC3 or a functional variants thereof, respectively.

[0163] In some embodiments, the cell or plurality of cells is a fibroblast comprising comprises one or a combination of ROBO2, EID1, CDH4, NCAM1, GPC3 or one or a combination of functional variants thereof that comprise at least about 75% sequence identity to ROBO2. EID1, CDH4, NCAM1, GPC3. In some embodiments, the cell or plurality of cells is afibroblast comprising comprises one or a combination of ROBO2, EID1, CDH4, NCAM1, GPC3, SOX9, and SOX10, or one or a combination of functional variants thereof that comprise at least about 75% sequence identity to ROBO2, EID1, CDH4, NCAM1, GPC3, SOX9, and SOX10. In some embodiments, the cell or plurality of cells is a fibroblast comprising comprises one or a combination of ROBO2, EID1, SOX9, and SOX10, or one or a combination of functional variants thereof that comprise at least about 75% sequence identity to ROBO2, EID1, SOX9, and SOX10. In some embodiments, the disclosure provides a composition comprising a cell or plurality of cells, wherein the cell or plurality of cells are derived from a stem cell or a human fibroblast cell line, such as 293T cells. In some embodiments, the stem cell is an induced pluripotent stem cell. In some embodiments, the cell or plurality of cells arehuman. In some embodiments, the disclosure provides a pharmaceutical composition comprising a therapeutically affective amount of any of the embodiments above and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides a kit comprising: (a) the composition or pharmaceutical compositions of any cells disclosed herein; and

[0164] (b) tissue culture medium and / or tubing configured for transfer of cells. In some embodiments, the kit further comprises instructions to culture or transfer the cells. In some embodiments, the kit is frozen.

[0165] In some embodiments, pharmaceutical compositions of the disclosure include a pharmaceutical composition comprising (i) a therapeutically effective amount of a population of fibroblasts comprising one or a combination of ROBO2, EID1, CDH4, NCAM1, GPC3, SOX9, and SOXIO, or one or a combination of functional variants thereof that comprise at least about 75% sequence identity to ROBO2, EID1, CDH4, NCAM1, GPC3, SOX9, and SOXIO; and (ii) a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is free of or substantially free of profibrotic fibroblasts. In some embodiments, the pharmaceutical composition is free of or substantially free of fibroblasts deficient in SOX9 and / or SOXIO.Cells

[0166] The disclosure relates to cells and, specifically ectodermal progenitor cells and well as differentiated forebrain, midbrain, and hindbrain cells. In some embodiments, the cells are mammalian cells. In some embodiments, the cells are human. The disclosure also relates to transformed cells lines that are optionally exposed to cell culture medium. In some embodiments, cells of the disclosure comprise or express an amino acid sequence comprising one or a combination of two or more biomarkers disclosed in Table 1.

[0167] In some embodiments, cells of the disclosure comprise or express an amino acid sequence comprising one or a combination of variants or functional fragments of the biomarkers disclosed in Table 1. In such embodiments, the functional fragment may retain 99%. 98%. 97%. 96%. 95%. 94%. 93%. 92%. 91%. or 90% sequence identity to the wild-type human sequence upon which the sequence is derived. In some embodiments, the functional fragment may retain 85%, 80%, 75%, 70%, 65%, or 60% sequence identity to the wild-type sequence upon which the sequence is derived. In some embodiments, the functional fragment may retain about 99%. 98%. 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 85%, 80%, 75%, 70%, 65%, or about 60% sequence identity to the amino acid sequence of Table 1.

[0168] Compositions of the disclosure include embodiments that comprise cells comprising ROBO2 or variants or functional fragments thereof that comprise no less than about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity to ROBO2.

[0169] In some embodiments cells of the disclosure comprise SEQ ID NO:4 or variants or functional fragments thereof that possess no less than about 70%. about 75%, about 80%, about 85%. about 90%. about 95%, about 96%, about 97%, about 98%, or about 99% sequence identity, to SEQ ID NO:4.

[0170] In some embodiments the cells comprise biologically active SOX9 and / or biologically active SOX 10. In some embodiments, the cells comprise a phenotype o?ROBO2 ' + Cdh4+. Ncaml+. Gpc3, but are deficient in biologically active SOX9 and / or SOX10.SOX-9 [Homo sapiens]NCBI Reference Sequence: NP_000337.1MNLLDPFMKMTDEQEKGLSGAPSPTMSEDSAGSPCPSGSGSDTENTRPQE NTFPKGEPDLKKESEEDKFPVCIREAVSQVLKGYDWTLVPMPVRVNGSSK NKPHVKRPMNAFMVWAQAARRKLADQYPHLHNAELSKTLGKLWRLLNESE KRPFVEEAERLRVQHKKDHPDYKYQPRRRKSVKNGQAEAEEATEQTHISP NAIFKALQADSPHSSSGMSEVHSPGEHSGQSQGPPTPPTTPKTDVQPGKA DLKREGRPLP EGGRQPPIDF RDVDIGELSS DVISNIETFD VNEFDQYLPP NGHPGVPATHGQVTYTGSYGISSTAATPASAGHVWMSKQQAPPPPPQQPP QAPPAPQAPPQPQAAPPQQPAAPPQQPQAHTLTTLSSEPGQSQRTHIKTE QLSPSHYSEQQQHSPQQIAYSPFNLPHYSPSYPPITRSQYDYTDHQNSSSYYSHAAGQGTGLYSTFTYMNPAQRPMYTPIADTSGVPSIPQTHSPQHWEQ PVYTQLTRPSOX- 10 [Homo sapiens]

[0171] MAEEQDLSEVELSPVGSEEPRCLSPGSAPSLGPDGGGGGSGLRASPGPGEL GKVKKEQQDGEADDDKFPVCIREAVSQVLSGYDWTLVPMPVRVNGASKSKPHVKRP MNAFMVWAQAARRKLADQYPHLHNAELSKTLGKLWRLLNESDKRPFIEEAERLRM QHKKDHPDYKYQPRRRKNGKAAQGEAECPGGEAEQGGTAAIQAHYKSAHLDHRHP GEGSPMSDGNPEHPSGQSHGPPTPPTTPKTELQSGKADPKRDGRSMGEGGKPHIDFG NVDIGEISHEVMSNMETFDVAELDQYLPPNGHPGHVSSYSAAGYGLGSALAVASGHS AWISKPPGVALPTVSPPGVDAKAQVKTETAGPQGPPHYTDQPSTSQIAYTSLSLPHYGSAFPSISRPQFDYSDHQPSGPYYGHSGQASGLYSAFSYMGPSQRPLYTAISDPSPSGPQ SHSPTHWEQPVYTTLSRP

[0172] The disclosure relates to an isolated fibroblast comprising, or cell line comprising a fibroblast that comprises biologically active ROBO2 or a functional variant thereof. In some embodiments, the isolated fibroblast comprises one or a combination of two or more of: Cdh4+. Ncaml+. Gpc3 or functional fragments thereof that comprise about 75%, 80%, 85%, 90%. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequences below.

[0173] Human Cadhenn 4 (CDH4), NCBI Ref No. NM_001794.5, is as follows:MTAGAGVLLLLLSLSGALRAHNEDLTTRETCKAGFSEDDYTALISQNILEGEKLLQVK FSSCVGTKGTQYETNSMDFKVGADGTVFATRELQVPSEQVAFTVTAWDSQTAEKWD AVVRLLVAQTSSPHSGHKPQKGKKVVALDPSPPPKDTLLPWPQHQNANGLRRRKRD WVIPPINVPENSRGPFPQQLVRIRSDKDNDIPIRYSITGVGADQPPMEVFSIDSMSGRM YVTRPMDREEHASYHLRAHAVDMNGNKVENPIDLYIYVIDMNDNRPEFINQVYNGS VDEGSKPGTYVMTVTANDADDSTTANGMVRYRIVTQTPQSPSQNMFTINSETGDIVTVAAGLDREKVQQYTVIVQATDMEGNLNYGLSNTATAIITVTDVNDNPPEFTASTFAGE VPENRVETVVANLTVMDRDQPHSPNWNAVYRIISGDPSGHFSVRTDPVTNEGMVTVVKAVDYELNRAFMLTVMVSNQAPLASGIQMSFQSTAGVTISIMDINEAPYF PSNHKLIRLEEGVPPGTVLTTFSAVDPDRFMQQAVRYSKLSDPASWLHINATNGQITTA AVLDRESLYTKNNVYEATFLAADNGIPPASGTGTLQIYLTDINDNAPELLPKEAQICE KPNLNAINITAADADVDPNIGPYVFELPFVPAAVRKNWTITRLNGDYAQLSLRILYLE AGMYDVPIIVTDSGNPPLSNTSIIKVKVCPCDDNGDCTTIGAVAAAGLGTGAIVAILICILILLTMVLLFVMWMKRREKERHTKQLLIDPEDDVRDNILKYDEEGGGEEDQDYD LSQLQQPEAMGHVPSKAPGVRRVDERPVGAEPQYPIRPMVPHPGDIGDFINEGLRAA DNDPTAPPYDSLLVFDYEGSGSTAGS VS SENS S S S GDQDYDYLNDWGPRFKKL ADM YGGGEED.

[0174] The amino acid sequence of human NCAM1, Accession Number NP_001387541.1, is as follows:1 mlqtkdli t Ifflgtavsl qvdivpsqge isvgeskffl cqvagdakdk diswfspnge61 kltpnqqris vvwnddssst Itiynanidd agiykcvvtg edgseseatv nvkifqklmf 121 knaptpqefr egedavivcd vvsslpptii wkhkgrdvil kkdvrfivls nnylqirgik 181 ktdegtyrce grilargein fkdiqvivnv pptiqarqni vnatanlgqs vtlvcdaegf 241 peptmswtkd geqieqeedd ekyifsddss qltikkvdkn deaeyiciae nkageqdati301 hlkvfakpki tyvenqtame leeqvtltce asgdpipsit wrtstmiss eekaswtrpe361 kqevhapwnw qvgrqkgqag sagfpgshet Idghmvvrsh arvssltlks iqytdageyi421 ctasntigqd sqsmylevqy apklqgpvav ytwegnqvni tcevfaypsa tiswfrdgql481 Ipssnysnik iyntpsasyl evtpdsendf gnynctavnr igqeslefil vqadtpssps541 idqvepysst aqvqfdepea tggvpilkyk aewravgecv whskwydake asmegivtiv601 glkpettyav rlaalngkgl geisaasefk tqpvhspppp asassstpvp Isppdttwpl661 palatepar epsapklegq mgedgnsikv nlikqddggs pirhylvryr alssewkpei721 rlpsgsdhvm Iksldwnaey evywaenqq gkskaahfvf rtsaqptaip atlggnsasy781 tfvsllfsav tllllc

[0175] The amino acid sequence of human glypican 3 [ Homo sapiens (human) ](GPC3), Acession No. KX533474.1, is as follows:MAGTVRTACLVVAMLLSLDFPGQAQPPPPPPDATCHQVRSFFQRLQPGLKWVPETPVPGSDLQVCLPKGPTCCSRKMEEKYQLTARLNMEQLLQSASMEL KFLIIQNAAVFQEAFEIVVRHAKZNYTNAMFKNNYPSLTPQAFEFVGEFFTDVSLYILGS DINVDDMVNELFDSLFPVIYTQLMNPGLPDSALDINECLRGARRDLKVFGNFPKLIM TQVSKSLQVTRIFLQALNLGIEVINTTDHLKFSKDCGRMLTRMWYCSYCQGLMMVK PCGGYCNVVMQGCMAGVVEIDKYWREYILSLEELVNGMYRIYDMENVLLGLFSTIH DSIQYVQKNAGKLTTTIGKLCAHSQQRQYRSAYYPEDLFIDKKVLKVAHVEHEETLSSRRRELIQKLKSFISFYSALPGYICSHSPVAENDTLCWNGQELVERYSQKAARNGMKN QFNLHELKMKGPEPVVSQIIDKLKHINQLLRTMSMPKGRVLDKNLDEEGFESGDCGD DEDECIGGSGDGMIKVKNQLRFLAELAYDLDVDDAPGNSQQATPKDNEISTFHNLGN VHSPLKLLTSMAISVVCFFFLVH

[0176] The isolated fibroblast can be one or a plurality of cells in a cell line. Generally, many techniques are known to isolate and propagate cells and develop cell lines. Briefly, somatic cells transfected with retroviral vectors that express OCT4, SOX2, KLF4 and cMYC to generate induced pluripotent stem cells ("iPSCs") express the same pluripotency markers as control H9 ESCs.

[0177] A subpopulation of human dermal fibroblasts that express the marker ROBO2 demonstrates enhanced iPSC generation efficiency. ROBO2 -positive and ROBO2-negative populations can be transduced with the same retroviral vectors, under identical experimental conditions, and seeded onto inactivated mouse embryonic fibroblasts (MEFs). After three weeks of culture under standard hESC conditions, plates can be examined in a double-blind analysis by three independent hESC biologists for iPSC colony formation. All three biologicalreplicates with the transduced R0B02 -negative cells can form many large background colonies but no iPSC colonies are likely to emerge. In contrast, all three biological replicates with the transduced R0B02 -positive cells can result in the formation of iPSC colonies (4-5 per replicate) but very few large background colonies (0-1 per replicate). Further characterization of the cell lines derived from the iPSC-like colonies showed that they possessed hESC-like morphology, growing as flat colonies with large nucleo-cytoplasmic ratios, defined borders and prominent nucleoli. When five lines were further expanded and characterized, all demonstrated expression of key pluripotency markers expressed by hESCs, which included alkaline phosphatase, Nanog, ROBO2, Cadherin 4, GPC3, NC AMI . If no iPSC colony formation or line derivation from the transduced ROBO2 -negative cells was observ ed, this indicates that these cells possess significantly lower or even no reprogramming potential relative to the ROBO2-expressing cells.

[0178] Additionally, a 10-fold enrichment of primary fibroblasts that strongly express ROBO2 results in a significantly greater efficiency (8-fold increase) of iPSC line derivation compared to the control derivation rate (p<0.05).

[0179] The ROBO2 -positive cells can be assessed morphologically, from the ROBO2 - negative fibroblasts; furthermore, expression of the ROBO2 antigen is not considered a marker of other cell ty pes such as mesenchymal or epidermal adult stem cells.

[0180] A rare subpopulation of ROBO2 expressing cells can be isolated that exists in the dermis of adult human skin. These ROBO2-expressing cells undergo a significant increase in cell number in response to injury, indicating a role in regeneration. These ROBO2-expressing cells were derived through primary cell culture, purified by fluorescence activated cell sorting (FACS) and characterized. The cells can be assessed for a global transcriptional state most similar to bone marrow and fat derived mesenchymal stem cells (MSCs) and the highest expressing ROBO2 expressing cells and can optionally co-express Cadherin 4, GPC3, and / or NCAM1. These cells represent a population for use in skin repair or regeneration. In some embodiments, the cells are mammalian cells. In some embodiments, the cells are human.Methods

[0181] The disclosure relates to methods of preventing or methods of treating fibrosis in a subject in need thereof comprising administering to the subject an effective amount of the pharmaceutical composition disclosed herein. In some embodiments, the pharmaceutical composition comprises one or a plurality of fibroblasts that are SOX9+ and SOX10+. In someembodiments, the method comprises a step of administering a second agent that has a therapeutic active agent that treats or prevents fibrosis.

[0182] In some embodiments, the methods are free of administration of second active agent and the step of administering consists of administering a single active agent in a therapeutically effective amount.

[0183] The disclosure relates to a method of inducing wound healing in a subject in need thereof comprising administering to the subject a composition comprising a EP300 modulator or any of the disclosed compositions, including composition comprising cells. In some embodiments, the EP300 modulator is chosen from any of the disclosed E300 inhibitors, or a pharmaceutically acceptable salt thereof or a prodrug thereof. In some embodiments, the EP300 modulator is chosen from: EP300i, Y08197, CCS1477. CPI-1612, DS17701585, DS- 9300, or NEO3734, or a pharmaceutically acceptable salt thereof, analog thereof or a derivative thereof. In some embodiments, the methods of the disclosure relate to administering to the subject a composition comprising an effective amount of a EP300 modulator and an effective amount of an opioid. Methods of the disclosure also relate to a method of potentiating an effect of an opioid in a subject in need thereof, the method comprising administering to the subject one or more of the compositions disclosed herein.

[0184] The disclosure relates to a method of treating fibrosis and methods of preventing fibrosis of a wound in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising an amount of from about 0.1 mg / kg / day to about 100 mg / kg / day of a EP300 modulator. In some embodiments, the method comprises a step of administering to the subject a pharmaceutical composition comprising an amount of from about 0.2 mg / kg / day to about 100 mg / kg / day of a EP300 modulator. In some embodiments, the method comprises a step of administering to the subject a pharmaceutical composition comprising an amount of from about 0.3 mg / kg / day to about 100 mg / kg / day of a EP300 modulator. In some more particular embodiments, methods, as provided herein, comprise administering a compound of any formula disclosed herein or a pharmaceutically acceptable salt thereof, to a subject twice a day, daily, every’ other day. three times a week, twice a week, weekly, every other week, twice a month, or monthly. In some embodiments, the method comprises administering a composition comprising a therapeutically effective amount of a polypeptide disclosed herein, or a pharmaceutically acceptable salt thereof, or a variant thereof to a subject daily.

[0185] The disclosure also relates to a method of preventing fibrosis and simultaneously accelerating wound healing in a subject in need thereof comprising administering to a subjectin need thereof a pharmaceutical composition comprising an effective amount of an EP300 modulator, or administering to the subject a first pharmaceutical composition comprising an EP300 modulator and a second pharmaceutical composition comprising a fibrosis treatment. In some embodiments, the fibrosis treatment is one more treatments chosen from: vitamin E, retinol, silicone sections or sheets, allantoin, bleomycin, and a corticosteroid.

[0186] In some embodiments, the EP300 modulator is administered over a period of about 3 days to about 12 months. In some embodiments, the EP300 modulator is administered during a surgical procedure, and / or once within about one, two, three, four, five, six, seven, eight, nine or fewer hours within the subject having a surgical procedure. In some embodiments, the method comprises administering the EP300 modulator once per day for about 1. 2, 3, 4. 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 or more days.

[0187] The disclosure relates to methods of promoting wound healing in a subject in need thereof comprising administering to the subject an effective amount of the pharmaceutical composition comprising an EP300 modulator or one or a plurality of cells disclosed herein.

[0188] The disclosure also relates to methods of reducing severity of scarring or preventing scarring in a subject comprising administering to the subject an effective amount of the pharmaceutical composition comprising an EP300 modulator or one or a plurality of cells disclosed herein.

[0189] The disclosure relates to methods of enhancing SOX10 and / or SOX9 expression in a wound of a subject in need comprising administering to a wound of the subject an effective amount of the pharmaceutical composition comprising an EP300 modulator or one or a plurality of cells disclosed herein.

[0190] The disclosure also relates to methods of inhibiting acetyltransferase in a wound of a subject in need thereof comprising administering to the subject an effective amount of the pharmaceutical composition comprising an EP300 modulator or one or a plurality of cells disclosed herein.

[0191] The disclosure also relates to methods of reducing a population of profibrotic fibroblasts in a subject in need thereof comprising administering to the subject an effective amount of the pharmaceutical composition comprising an EP300 modulator or one or a plurality of cells disclosed herein.

[0192] Suitable dosage ranges for intravenous (i.v.) administration are about 0.01 milligram to aboutlOO milligrams per kilogram body weight, about 0.1 milligram to about 35 milligrams per kilogram body weight, and 1 milligram to 10 milligrams per kilogram body weight. Suitable dosage ranges for intranasal administration are generally about 0.01 ng / kgbody weight to 1 mg / kg body weight. Suppositories generally contain 0.01 milligram to 50 milligrams of a compound of the disclosure per kilogram body weight and comprise active ingredient in the range of about 0.5% to about 10% by weight. Recommended dosages for intradermal, intramuscular, intraperitoneal, subcutaneous, epidural, sublingual, intracerebral, intravaginal, transdermal administration or administration by inhalation are in the range of about 0.001 milligrams to about 200 milligrams per kilogram of body weight. Suitable doses of the compounds of the disclosure for topical administration are in the range of 0.001 milligram to 1 milligram, depending on the area to which the compound is administered. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems. Such animal models and systems are well known in the art.

[0193] The disclosure also provides pharmaceutical packs or kits comprising one or more containers filled with one or more compounds of the disclosure. Optionally associated with such container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration. In a certain embodiment, the kit contains more than one compound of the disclosure. In some embodiments, the kit comprises a composition comprising an EP300 modulator and a composition comprising a second active agent. In some embodiments, the EP300 modulator is a EP300 modulator selected from any polypeptide disclosed herein or variants thereof, a pharmaceutically acceptable salt thereof, and a prodrug thereof. In some embodiments, the EP300 modulator is chosen from: EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS- 9300, or NEO3734, or a pharmaceutically acceptable salt thereof, analog thereof or a derivative thereof. In some embodiments, kit comprises a container containing frozen cells disclosed herein and instructions on how to culture and propagate such cells. In some embodiments, the kit comprises a container comprising a pharmaceutical composition disclosed here and, optionally, a syringe or device for administration of the pharmaceutical composition.

[0194] The compounds of the disclosure can be assayed in vitro and in vivo, for the desired therapeutic or prophylactic activity, prior to use in humans. For example, in vitro assays can be used to determine whether administration of a specific compound of the disclosure or a combination of compounds of the disclosure is for treating fibrosis or promoting wound healing. The compounds of the disclosure may also be demonstrated to be effective and safe using animal model systems.

[0195] The disclosure relates to a method of culturing fibroblast, plurality’ of fibroblasts or any cells disclosed herein. In some embodiments, method of culturing comprises (a) exposinga fibroblast to one or a combination of EP300 modulators for about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 24. 36 or about 48 hours. In some embodiments, the method comprises culturing a differentiated fibroblast disclosed herein for a time period sufficient to differentiate the cell or cells into expressing SOX9 and / or SOXIO or any functional variants of the foregoing. In some embodiments, the fibroblasts cells are cultured or exposed to an EP300 modulator for a time period sufficient to differentiate the fibroblast into exhibiting a phenotype comprising ROBO2, EID1, CDH4, NCAM1. GPC3 or one or a combination of functional variants thereof that comprise at least about 75% sequence identity to ROBO2, EID1, CDH4, NCAM1, GPC3. In some embodiments, the fibroblasts cells are cultured or exposed to an EP300 modulator for a time period sufficient to differentiate the fibroblast into exhibiting a phenotype comprising comprises one or a combination of ROBO2, EID1, CDH4, NCAM1. GPC3, SOX9, and SOXIO, or one or a combination of functional variants thereof that comprise at least about 75% sequence identity to ROBO2, EID1, CDH4, NCAM1, GPC3, SOX9, and SOXIO. In some embodiments, the fibroblasts cells are cultured or exposed to an EP300 modulator for a time period sufficient to differentiate the fibroblast into exhibiting a phenotype comprising ROBO2, EID1, SOX9, and SOXIO. or one or a combination of functional variants thereof that comprise at least about 75% sequence identity to ROBO2, EID1, SOX9, and SOXIO. In some embodiments, the methods of the disclosure comprise a method of differentiating a cell into a fibroblast disclosed herein comprising one or a combination of ROBO2, EID1, CDH4, NCAM1. GPC3 or any functional variant thereof. In some embodiments, the methods of culturing and differentiating the cell or cells are exposed to a media comprising an EP300 modulator disclosed herein.

[0196] In some embodiments, the method further comprises exposing the cell or cells to medium comprising RPM1, DMEM for about 18 hours, 19 hours, 20, hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hour, 28 hours, 29 hours, 30, 31, 32, 33, 34, 35, 36 hours or more of time. In some embodiments, the method further comprises differentiating the stem cell into a fibroblast that is nonfibrotic. In some embodiments, the method further comprises a step of trypsinizing the cells from a solid support such as a plastic surface and spinning the cells before aliquoting the cells into a separate container. In some embodiments, the cell compositions are manufactured by aliquoting from about 1 million to about 100 million cells into a container comprising PBS or other neutral buffer. Compositions of the disclosure include pharmaceutical compositions comprising aliquots of cells suspended in sterile and neutralized phosphate buffer. In some embodiments, the pH of the solution suspending the disclosed cells is from about 6 to about 7.9.

[0197] Although the disclosure has been described with reference to exemplary embodiments, it is not limited thereto. Those skilled in the art will appreciate that numerous changes and modifications may be made to the preferred embodiments of the disclosure and that such changes and modifications may be made without departing from the true spirit of the disclosure. It is therefore intended that the appended claims be construed to cover all such equivalent variations as fall within the true spirit and scope of the disclosure. All referenced journal articles, patents, and other publications disclosed herein are incorporated by reference herein in their entireties.REFERENCES

[0198] All references cited herein, below or otherwise, are incorporated herein by reference as if fully set forth.1. Henderson, N. C , Rieder, F. & Wynn, T. A. Fibrosis: from mechanisms to medicines. Nature 587, 555-566, doi: 10.1038 / s41586-020-2938-9 (2020).2. Brockes, J. P., Kumar, A. & Velloso, C. P. Regeneration as an evolutionary’ Brockes, J. P., Kumar, A. & Velloso, C. P. Regeneration as an evolutionary.3. Ferguson. M. W. & O'Kane, S. Scar-free healing: from embryonic mechanisms to adult therapeutic intervention. Philos Trans R Soc Land B Biol Sci 359, 839-850, doi: 10. 1098 / rstb.2004. 1475 (2004).4. Marshall, C. D. el al. Cutaneous Scarring: Basic Science, Current Treatments, and Future Directions. Adv Wound Care (New Rochelle) 7,29-45. doi: 10.1089 / wound.2016.0696 (2018).5. Forsch, R. T. Essentials of skin laceration repair. Am Bam Physician 78, 945-951 (2008).6. Gauglitz, G. G., Korting, H. C., Pavicic, T., Ruzicka, T. & Jeschke, M. G. Hypertrophic scarring and keloids: pathomechanisms and current and emerging treatment strategies. Mol Med 17, 113-125, doi: 10.2119 / molmed.2009.00153 (2011).7. Aarabi, S. et al. Mechanical load initiates hypertrophic scar formation through decreased cellular apoptosis. FASEB J 21, 3250-3261. doi:10. 1096 / fi.07-8218com (2007).8. Edlich, R. F. & Carl, B. A. Predicting scar formation: from ritual practice (Langer's lines) to scientific discipline (static and dynamic skin tensions). J Emerg Med 16. 759-760. doi: 10.1016 / s0736-4679(98)00070-5 (1998).9. Wong, V. W., Levi, K., Akaishi, S., Schultz, G. & Dauskardt, R. H. Scar zones: regionspecific differences in skin tension mav determine incisional scar formation. Plast Reconstr Surg 129, 1272-1276, doi:10.1097 / PRS.0b013e31824eca79 (2012).10. Rinkevich, Y. et al. Skin fibrosis. Identification and isolation of a dermal lineage withintrinsic fibrogenic potential. Science 348, aaa2151, doi:10. 1126 / science.aaa2151 (2015).11. Mascharak, S. et al. Preventing Engrailed-1 activation in fibroblasts yields wound regeneration without scarring. Science 372, doi: 10.1126 / science.aba2374 (2021).12. Chang, H. Y. et al. Diversity, topographic differentiation, and positional memory in human fibroblasts. Proc Natl Acad Sci U S A 99, 12877-12882, doi: 10. 1073 / pnas. 162488599 (2002).13. Rinn, J. L. et al. A dermal HOX transcriptional program regulates site-specific epidermal fate. Genes Dev 22, 303-307, doi: 10.1101 / gad.l610508 (2008).14. Myung, P., Andi, T. & Atit, R. The origins of skin diversity': lessons from dermal fibroblasts. Development 149, doi: 10.1242 / dev.200298 (2022).15. Leavitt. T. et al. Prrxl Fibroblasts Represent a Pro-fibrotic Lineage in the Mouse Ventral Dermis. Cell Rep 33, 108356, doi:10.1016 / j.celrep.2020.108356 (2020).16. Jiang, X., Iseki, S., Maxson, R. E., Sucov, H. M. & Morriss-Kay, G. M. Tissue origins and interactions in the mammalian skull vault. Dev Biol 241, 106-116 (2002).17. LaBonne, C. & Bronner-Fraser, M. Molecular mechanisms of neural crest formation. Anna Rev Cell Dev Biol 15, 81-112, doi: 10. 1146 / annurev. cellbio. 15. 1.81 (1999).18. Zalc. A. et al. Reactivation of the pluripotency program precedes formation of the cranial neural crest. Science 371, doi: 10.1126 / science.abb4776 (2021).19. Selleri, L. & Rijli, F. M. Shaping faces: genetic and epigenetic control of craniofacial morphogenesis. Nat Rev Genet, doi: 10. 1038 / s41576-023-00594 w (2023).20. Crane, J. F. & Trainor, P. A. Neural crest stem and progenitor cells. Annu Rev Cell Dev Biol 22, 267-286, doi: 10.1146 / annurev.cellbio.22.010305. 103814 (2006).21. Candido-Ferreira, I. L., Lukoseviciute, M. & Sauka-Spengler, T. Multi-layered transcriptional control of cranial neural crest development. Semin Cell Dev Biol 138. 1-14. doi : 10. 1016 / j . semcdb.2022.07.010 (2023).22. Ransom, R. C. et al. Mechanoresponsive stem cells acquire neural crest fate in jaw regeneration. Nature 563, 514-521, doi: 10.1038 / s41586-018-0650-9 (2018).23. D Bellard. M. E., Rao. Y. & Bronner-Fraser, M. Dual function of Slit2 in repulsion and enhanced migration of trunk, but not vagal, neural crest cells. J Cell Biol 162, 269-279, doi: 10. 1083 / jcb.200301041 (2003).24. Jia, L., Cheng, L. & Raper, J. Slit / Robo signaling is necessary' to confine early neural crest cells to the ventral migratory pathway in the trunk. Dev Biol 282, 411-421, doi: 10.1016 / j.ydbio.2005.03.021 (2005).25. Li. Y. et al. Robo signaling regulates the production of cranial neural crest cells. Exp Cell Res 361, 73-84, doi: 10.1016 / j.yexcr.2017. 10.002 (2017).26. Mascharak, S. et al. Multi-omic analysis reveals divergent molecular events in scarring and regenerative wound healing. Cell Stem Cell 29, 315-327 e316,doi : 10.1016 / j . stem.2021.12.011 (2022).27. Gulati, G. S. et al. Single-cell transcriptional diversity is a hallmark of developmental potential. Science 367. 405-411, doi: 10.1126 / science.aax0249 (2020).28. Kelsh, R. N. Sorting out SoxlO functions in neural crest development. Bioessays 28, 788-798, doi: 10.1002 / bies.20445 (2006).29. Li, J., Geraldo, L. H., Dubrac, A., Zarkada, G. & Eichmann, A. Slit2- RoboSignaling Promotes Glomerular Vascularization and Nephron Development. J Am Soc Nephrol 32, 2255-2272, doi:10.1681 / ASN.2020111640 (2021).30. MacLellan, W. R., Xiao, G., Abdellatif, M. & Schneider, M. D. A novel Rb- and p300- binding protein inhibits transactivation by MvoD. Mol Cell Biol 20, 8903-8915, doi: 10. 1128 / MCB.20.23.8903-8915.2000 (2000).31. Miyake, S. et al. Cells degrade a novel inhibitor of differentiation with ElA-like properties upon exiting the cell cvcle. Mol Cell Biol 20. 8889-8902, doi: 10.1128 / MCB.20.23.8889-8902.2000 (2000).32. Visel, A. et al. ChlP-seq accurately predicts tissue-specific activity of enhancers. Nature 457, 854-858, doi: 10. 1038 / nature07730 (2009).33. Fu, X. et al. Transcriptome profiling in Eidl-KO mice brain shows that Eidl links cell proliferation in the brain. Gene 717, 143998, doi: 10. 1016 / j.gene.2019.143998 (2019).34. Fu, X. et al. EID1 plays a crucial role in proliferation of neural stem cell. Biochem Biophys Res Commun 512, 763-769, doi: 10.1016 / j.bbrc.2019.03.138 (2019).35. Chen, X. et al. Protein Palmitoylation Regulates Neural Stem Cell Differentiation by Modulation of EID1 Activity. Mol Neurobiol 53, 5722-5736. doi: 10.1007 / sl2035-015- 9481-y (2016).36. Shahbazi, J. et al. The Bromodomain Inhibitor JQ1 and the Histone Deacetylase Inhibitor Panobinostat Synergistically Reduce N-Mvc Expression and Induce Anticancer Effects. Clin Cancer Res 22, 2534-2544, doi: 10.1158 / 1078-0432.CCR-15-1666 (2016).37. Gokani, S. & Bhatt. L. K. Bromodomains: A novel target for the anticancer therapy. Eur J Pharmacol 911, 174523, doi:10.1016 / j.ejphar.2021.174523 (2021).38. Picaud, S. et al. Generation of a Selective Small Molecule Inhibitor of the CBP / p300 Bromodomain for Leukemia Therapy. Cancer Res 75, 5106-5119, doi: 10. 1158 / 0008-5472. CAN-15-0236 (2015).39. Cheung, M. & Briscoe, J. Neural crest development is regulated by the transcription factor Sox9. Development 130, 5681-5693. doi: 10.1242 / dev.00808 (2003).40. Yao, T. P. et al. Gene dosage-dependent embryonic development and proliferation defects in mice lacking the transcriptional integrator p300. Cell 93. 361-372, doi : 10.1016 / s0092-8674(00)81165 -4 ( 1998).41. Ross, A. P. & Zarbalis, K. S. The emerging roles of ribosome biogenesis in craniofacial development. Front Physiol 5, 26, doi: 10.3389 / fphys.2014.00026 (2014).42. Foster, D. S. et al. Integrated spatial multiomics reveals fibroblast fate during tissue repair. Proc Natl Acad Sci U S A 118, doi: 10.1073 / pnas.2110025118 (2021).43. Griffin. M. F. et al. Piezo inhibition prevents and rescues scarring by targeting the adipocyte to fibroblast transition. bioRxiv, doi: 10. 1101 / 2023.04.03.535302 (2023).44. Wong, V. W. et al. Focal adhesion kinase links mechanical force to skin fibrosis via inflammatory signaling. Nat Med 18, 148-152, doi: 10.1038 / nm.2574 (2011).45. Talbott, H. E., Mascharak, S., Griffin, M., Wan, D. C. & Longaker, M. T. Wound healing, fibroblast heterogeneity', and fibrosis. Cell Stem Cell 29, 1161-1180, doi: 10. 1016 / j.stem.2022.07.006 (2022).46. Plikus, M. V. et al. Fibroblasts: Origins, definitions, and functions in health and disease. Cell 184, 3852-3872, doi: 10.1016 / j. cell.2021 .06.024 (2021).47. Griffin, M. F., Huber, J., Evan, F. J., Quarto, N. & Longaker, M. T. The role of Wnt signaling in skin fibrosis. Med Res Rev 42, 615-628, doi: 10. 1002 / med.21853 (2022).48. Black, S. et al. CODEX multiplexed tissue imaging vith DNA-conjugated antibodies. Nat Protoc 16, 3802-3835, doi: 10.1038 / s41596-021-00556-8 (2021).49. Bannister, A. J. & Kouzarides. T. Regulation of chromatin by histone modifications. Cell Res 21, 381-395, doi: 10.1038 / cr.2011.22 (2011).50. Creyghton, M. P. et al. Histone H3K27ac separates active from poised enhancers and predicts developmental state. Proc Natl Acad Set U SA 107, 21931- 21936, doi : 10. 1073 / pnas. 1016071 107 (2010).51. Kaya-Okur, H. S., Janssens. D. H., Henikoff. J. G., Ahmad, K. & Henikoff, S. Efficient low-cost chromatin profiling with CUT&Tag. Nat Protoc 15, 3264-3283. doi: 10.1038 / s41596-020-0373-x (2020).52. Skene, P. J. & Henikoff, S. An efficient targeted nuclease strategy for high- resolution mapping of DNA binding sites. Elife 6 doi: 10.7554 / eLife.21856 (2017).53. Dickinson, R. E. & Duncan, W. C. The SLIT-ROBO pathway: a regulator of cell function with implications for the reproductive system. Reproduction 139, 697- 704. doi: 10.1530 / REP-10-0017 (2010).54. Phesse, T. J. & Sansom. O. J. Responding to R-spondin: slit2 potentiates intestinal regeneration. Cell Stem CelllS, 512-514, doi: 10.1016 / j.stem.2013.10.006 (2013).55. Park, J. S., Cho, R., Kang, E. Y. & Oh, Y. M. Effect of Slit / Robo signaling on regeneration in lung emphysema. Exp Mol Med 53, 986-992, doi: 10.1038 / sl2276-021-00633-8 (2021).56. Pilling, D., Zheng, Z., Vakil, V. & Gomer, R. H. Fibroblasts secrete Slit2 to inhibit fibroevte differentiation and fibrosis. Proc Natl Acad Sci U S A 111, 18291- 18296, doi: 10.1073 / pnas. 1417426112 (2014).57. Watanabe, K., Yumimoto, K. & Nakayama, K. I. FBXO21 mediates the ubiquitylation and proteasomal degradation of EID 1. Genes Cells 20, 667-674,doi: 10.1111 / gtc. 12260 (2015).58. Conery, A. R. et al. Bromodomain inhibition of the transcriptional coactivators CBP / EP300 as a therapeutic strategy to target the IRF4 network in multiple myeloma. Elife 5, doi: 10.7554 / eLife.10483 (2016).59. Tang, X. et al. Assessment of Brd4 inhibition in idiopathic pulmonary’ fibrosis lung fibroblasts and in vivo models of lung fibrosis. Am J Pathol 183, 470-479, doi: 10.1016 / j.ajpath.2013.04.020 (2013).60. Ding. N. et al. BRD4 is a novel therapeutic target for liver fibrosis. Proc Natl Acad Sci USA 112, 15713-15718, doi: 10.1073 / pnas,1522163112 (2015).61. Chen, E. Y. et al. Enrichr: interactive and collaborative HTML5 gene list enrichment analvsis tool. BMC Bioinformatics 14, 128. doi: 10.1186 / 1471-2105-14-128 (2013).62. Gerstner. N. et al. GeneTrail 3: advanced high-throughput enrichment analysis. Nucleic Acids Res 48, W515-W520, doi: 10.1093 / nar / gkaa306 (2020).63. Wolf, F. A., Angerer, P. & Theis, F. J. SCANPY: large-scale single-cell gene expression data analysis. Genome Biol 19, 15, doi: 10. 1186 / sl3059-017-1382-0 (2018).64. Trapnell, C. et al. The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells. Nat Biotechnol 32, 381-386, doi: 10.1038 / nbt.2859 (2014).65. Ruifrok, A. C. & Johnston, D. A. Quantification of histochemical staining by color deconvolution. Anal Quant Cytol Histol 23, 291-299 (2001).66. Yu, G., Wang. L. G. & He, Q. Y. ChlPseeker: an R / Bioconductor package for ChIP peak annotation, comparison and visualization. Bioinformatics 31, 2382-2383, doi : 10.1093 / bioinformatics / btv 145 (2015).EXAMPLES

[0199] Using a four-site wounding model in mice, it was observed that neural crest- derived facial wounds heal with less scarring than cephalic mesoderm-derived scalp wounds, lateral plate mesoderm-derived ventral wounds, and paraxial mesoderm-derived dorsal wounds. Single-cell RNA sequencing identified increased expression of Robo2 and downstream Eidl in neural-crest derived facial fibroblasts compared to fibroblasts from other sites. Subsequent fibroblast transplantation experiments showed that Robo2 and Eidl are required for facial fibroblasts’ intrinsic reduced fibrotic potential. This is maintained by the EIDl-mediated inhibition of EP300 histone acetyltransferase, leading to a more transcriptionally silent chromatin landscape, including around extracellular matrix genes. Mimicking EID1 ’s endogenous activity, both small-molecule and transgenic repression of EP300 in dorsal wounds promoted facial -like healing with reduced scarring.Overall, these data highlight the importance of the R0B02-EID1- EP300 signaling axis in facial wound healing and demonstrate our ability- to modulate the embryologically determined fibrogenic potential of fibroblasts as a therapeutic approach to minimize scar formation.EXAMPLE 1MATERIALS AND METHODSMouse strains

[0200] The following mouse strains were acquired from Jackson Laboratories (Bar Harbor, ME): B6 (C57BL / 6J, Stock: 000664), CollalCre-ERT (B6.Cg-Tg(Collal- cre / ERT2)lCrm / J, Stock: 016241), Sox9-CreERT (Stock: 035092), SoxlO-CreERT (Strain#27651), EP300fl / fl (Strain #:025168). mTmG(B6.129(Cg)- Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo / J Stock: 007676), and Ai9. Mice were housed at the Stanford University Comparative Medicine Pavilion per Stanford APLAC guidelines, under the supervision of the Veterinary- Service Center (VSC). ROSA26mTmG mice utilize a dual-fluorescence reporter system that irreversibly substitutes Tomato red fluorescent protein (RFP) with membrane-bound green fluorescent protein (GFP) after recombination. Sox9-CreERT;mTmG and Sox 10-CreERT;mTmG mice were generated by crossing Sox9-CreERT and SoxlO-CreERT mice with mTmG mice, respectively. Collal- CreERT;mTmG;EP300+ / - mice were generated by crossing CollalCre-ERT with mTmG to generate homozygous Collal-Cre-ERT;mTmG progeny, which were then crossed with EP300fl / fL Equal numbers of female and male mice ranging from the age of 8-12 weeks were allocated to experimental groups. Sample size for any given experiment is detailed in the figure legend.

[0201] Four- region murine wounding model

[0202] Prior to wounding mice were anesthetized (2% isoflurane), their dorsal hair was removed with depilatory cream, and the dorsal skin was prepped with alternating betadine and alcohol wipes. Full-thickness 2.5mm excisional wounds were created on the facial, scalp, ventral, and dorsal skin. Silicone rings were sutured (eight simple interrupted 8-0 nylon (Ethicon) sutures per ring) to all excisional wounds to stent the wound open and minimize wound contraction throughout the healing process. Following wounding, postoperative analgesia was accomplished with buprenorphine SR 0.05 mg / kg every four hours for three doses, and then as indicated. Wounds were dressed using 3M TegadermTransparent Film 1626w dressings (3M™, Cat: 1626W), and dressing changes took place every 48 hours under anesthesia. Wounds were harvested at post-operative day 7 and 14 by excising a >5mm margin around the wound border; unwounded skin was also harvested from face, scalp, ventral, and dorsal skin. For mice receiving treatments with EP300 bromodomain inhibitor, treatments consisted of a single administration of 250 pg I- CBP112 (Selleck Chem, Inc; Radnor, PA) at POD 0, via local intradermal injections into the wound edge; PBS was injected for vehicle controls.

[0203] Histology and immunohistochemistry

[0204] Samples for histology' were fixed by incubation in 10% neutral buffered formalin phosphate for 16 hours at 4 °C. The samples were washed with PBS, dehydrated through serial ethanol washes, cleared with xylene, infiltrated with paraffin through serial incubations, and embedded in paraffin. Sections were cut at a thickness of 8 pm and incubated at 37 °C overnight to affix sections to slides prior to staining. Sections for immunohistochemistry were soaked in 30% sucrose dissolved in PBS at 4°C. After one week, samples were removed from the sucrose solution and embedded as tissue blocks using Tissue Tek O.C.T. (Sakura Finetek, Torrance, CA) over dry ice and 100% ethanol to achieve rapid freezing. Frozen blocks were mounted on a Thermo Scientific CryoStar NX70 cry ostat, and 8 pm-thick sections w ere transferred to Superfrost / Plus adhesive slides (ThermoFisher Scientific™, Waltham, MA). Hematoxylin and eosin (Vector Laboratories, Cat:H-3502). Masson’s Trichrome (Abeam, abl50686). and Picrosirius Red (Abeam, abl50681) staining was performed per standard protocols from the manufacturers. For immunofluorescent staining, samples were cleared with xylene, re-hydrated, and treated with antigen retrieval buffer per established protocols (Abeam, ab970). Samples w ere then permeabilized in 0.25% Triton-X (15 minutes), blocked for 2 hours (Powerblock), and stained overnight with primary antibodies in O. lx Powerblock (COL1 (ab270993), COL3 (22734-1-AP), aSMA (ab5694)), VIM (ab92547), ROBO2 (ab244331), EID1 (abl80133), EP300 (ab275378), SOX10 (ab227680)). Samples were then washed 3 times with O.lx Powerblock, stained for 1 hr with secondary antibodies (Invitrogen), washed another 3 times, and finally mounted with Fluoromount-G containing DAPI. Slides were imaged using a Leica TCS SP8 confocal microscope. Fluorescence intensity was quantified using ImageJ softw are to quantify pixel densify of each antibody channel.

[0205] Mouse fibroblast and skin transplantation

[0206] For mouse fibroblast transplantations, full-thickness skin was harvested from the face, scalp, ventrum, and dorsum of GFP+ mice. Tissues were processed andfibroblasts were isolated as described in the Fluorescence activated cell sorting Methods section. Sorted fibroblasts from each region were then injected (100,000 cells / site) subcutaneously into the unwounded dorsal skin or at the wound edges of full-thickness dorsal excisional wounds of separate wildtype mice. After 14 days, full-thickness skin at the injection sites was harvested for downstream analyses. For mouse skin transplantations, Full thickness (10mm x 10mm) skin grafts were harvested from the face, scalp, ventrum and dorsum from RFP+ mice and embedded into subdermal space of the dorsum of wildtype mice. Following skin graft integration (at post operative day 10), a 2 mm incisional wound was made on the RFP+ skin grafts. After 14 days of healing, healed wounds were harvested as above and processed for histological and immunohistochemical analyses.

[0207] Fluorescence activated cell sorting

[0208] Facial, scalp, ventral, and dorsal mouse skin was harvested using dissecting scissors by separation along fascial planes. Next, the subcutaneous fat was trimmed with a scalpel, and the skin was rinsed in betadine, followed by 5 rinses in cold PBS. To achieve a cell suspension, the harvested skin was finely minced using sharp scissors, enzymatically digested (Liberase DL, 0.5 mg / mL, 1 hour), and filtered through a 40 mm nylon mesh. Fibroblasts were isolated from experimental mice via a previously reported FACS strategy .10Briefly, a lineage gate (Lin) for hematopoietic (CD45, Ter- 119), endothelial (CD31. Tie2), and epithelial (CD326, CD324) cell markers was used as a negative gate to isolate fibroblasts (Lin-) and non-fibroblasts (Lin+). ROBO2+ (ab244331), CDH4+( MA5-41143), NCAM1+ (14255-1-AP), GPC3+(MA5- 16368) and EP300+(ab275378) fibroblasts were isolated from the Lin- population using their respective antibodies.

[0209] Single-cell RNA sequencing and analysis

[0210] Full-thickness skin from the face, scalp, ventrum and dorsum of unwounded animals and of excisional wounds at POD 7 and 14 were harvested and mechanically digested with sharp dissecting scissors (n = 6 per condition). Tissue was added to an enzymatic digestion consisting of Collagenase II (ThermoFisher, Cat: 17101015) and IV (ThermoFisher, Cat: 17104019) in DMEM-F 12 (GIBCOTM, Fischer Scientific. Hampton, NH). Samples were added to an orbital shaker at 150 rpm for 90 minutes at 37°C. FACS buffer was added to quench the digest and samples were filtered through 70 pm cell strainers. Samples were then centrifuged at 1500 for 5 minutes at 4°C, resuspended in FACS buffer, filtered for a second time using 40 pm cell strainers, and cell numbers were quantified on a hemocytometer. Cells from each sample were tagged with hashtag oligos(HTOs) per the manufacturer’s protocol and then pooled. Quality control and single cell RNAseq were performed on unsorted cells using the lOx Chromium Single Cell platform (Single Cell 3’ v3, lOx Genomics, USA) at the Stanford Functional Genomics Facility (SFGF), Stanford University, Palo Alto (NextSeq 500 Illumina at 50,000 reads per cell). Base calls were converted to reads using the Cell Ranger (10X Genomics; version 3.1) implementation mkfastq and then aligned against the Cell Ranger mmlO reference genome, available at: http: / / cf.10xgenomics.com / supp / cell-exp / , using Cell Ranger’s count function with SC3Pv3 chemistry and 5,000 expected cells per sample. A maximum percent mitochondrial RNA cutoff of 15% was employed. Unique molecular identifiers (UMIs) from each cell barcode were retained for all downstream analysis, normalized with a scale factor of 10,000 UMIs per cell, and subsequently natural log transformed with a pseudocount of 1 using the R package Seurat (version 4.0.5). The first 15 principal components of the aggregated data were then used for uniform manifold approximation and projection (UMAP) analysis. Cell annotations were ascribed using SingleR (version 3.11) against the Mouse-RNAseq reference dataset, available at https: / / rdrr.io / github / dviraran / SmgleR / man / mouse.maseq.html. Cell-type marker lists were generated using Seurat’s native FindMarkers function with a log fold change threshold of 0.25 using the ROC test to assign predictive power to each gene. The 200 most highly ranked genes from this analysis for each cluster were used to perform gene set enrichment analysis in a programmatic fashion using EnrichR (version 2. 1).61

[0211] GeneTrail analysis

[0212] Using GeneTrail 3, an over-representation analysis was performed for each cell using the 500 most expressed protein-coding genes on gene sets from Gene Ontology.62P values were adjusted using the Benjamini-Hochberg procedure, and gene sets were required to have between 2 and 1,000 genes. Stacked violin plots were generated using the Scanpy package.63

[0213] Cyto TRACE analysis

[0214] CytoTRACE was used to compare differentiation states among fibroblasts (https: / / cytotrace.stanford.edu / ).27 This tool analyzes the number of uniquely expressed genes per cell, as well as other factors like distribution of mRNA content and number of RNA copies per gene, to calculate a score assessing the differentiation and developmental potential of each cell (lowest differentiation and highest developmental potential at 1; highest differentiation and lowest developmental potential at 0). Cells arethen ordered by their predicted differentiation status. CytoTRACE analysis was performed using default parameters for each fibroblast in the dataset.

[0215] Pseudotime analysis

[0216] Pseudotime analysis was performed using the Monocle 3 package in R (version 3 0.2.0).64Counts for individual cells were preprocessed using PCA with 15 dimensions following log-normalization. Dimensional reduction was performed using a UMAP reduction with min_dist = 0.5. n_neighbors = 30, and repulsion. strength = 2.0. Cells were then clustered using Monocle 3’s Louvain implementation with a resolution of le- 5. A principal graph was then learned from the reduced dimension space using reversed graph embedding with default parameters, and cell order selection was made from the two elements at either end of the trajectory. Pseudotime trajectory heatmaps were created using the Monocle 2 package in R.

[0217] Picrosirius red histologic analysis

[0218] We used a previously described image-processing algorithm to quantify ECM ultrastructure of Picrosirius red-stained tissue slices.26The algorithm profiles 294 ultrastructural features (e.g., fiber length, width, persistence, alignment, overall dimensionality) to provide a quantitative comparison of extracellular matrices. Each group (n = 10) was randomly imaged at 100 separate locations at 40x. Color deconvolution following previously described methods66was performed to characterize each stain byabsorbance in three RGB channels. Ortho-normal transformation was then used to determine each color’s contribution to the captured image. Red and green images were produced, representing mature and immature ECM fibers, and analyzed as separate groups. A Matlab script was used to analyze the images, including performing noise reduction, preferential selection for smooth regions with low variance, and ■‘skeletonization” of the fiber networks.

[0219] RT-qPCR

[0220] RNeasy mini kit (Qiagen LLC, Germantown MD) was used for RNA extraction for real time quantitative polymerase chain reaction (qPCR). Transcription was then performed using Moloney murine leukemia virus reverse transcriptase. An ABI Prism PCR7500 sequence detection system (Applied Biosystems, Waltham, MA) using TaqMan expression assay (ThermoFisher C A) was used for transcriptional level quantification.

[0221] ELISA of wound tissue supernatant

[0222] Full-thickness skin was harvested from unwounded skin at the respective sites and wounds at POD 14 after various treatments and mechanically digested with sharpdissecting scissors. Tissue preparation for ELISA was done using ab210579 kit. Tissue was placed in microfuge tubes and immersed in liquid nitrogen to snap-freeze and subsequently kept on ice. Tissue homogenization was performed by adding 300 pl / 5mg tissue of complete extraction buffer to the tube and mechanically homogenized using sharp dissecting scissors. Constant agitation for 2 hours at 4°C was maintained thereafter on an orbital shaker. Tissue homogenates were then centrifuged for 20 min at 13,000 x rpm at 4°C. Supernatant for each homogenate was then aliquoted to a fresh, chilled tube and stored at -80°C until used for ELISA. Tissue homogenate supernatant was tested for COLIal levels using the Abeam mouse / human enzy me linked immunoassay (ELISA) kits (Abeam, Cat: abl08785, ab210579, ab229389, USA) according to manufacturer's protocol. Standards and samples were placed into wells coated with COLIal-specific antibody. Enzyme-linked polyclonal antibody was then added to wells, followed by the addition of substrate. A spectrophotometer captured absorbances, and concentrations of protein target were calculated and modeled using Prism.

[0223] Mouse primary fibroblast cell culture, and EP300 inhibitor and SLIT2 treatments

[0224] Full-thickness skin samples (10mm x 10mm dimensions) were separately harvested from the face and dorsum of B6 mice and mechanically digested with sharp dissecting scissors (n = 6 per condition). Tissue was then added to an enzymatic digestion consisting of Collagenase II (ThermoFisher, Cat: 17101015) and IV (ThermoFisher, Cat: 17104019) in DMEM-F12 (GIBCOTM, Fischer Scientific, Hampton, NH). Samples were added to an orbital shaker at 150 rpm for 90 minutes at 37°C and FACS buffer was added to quench the digest. Samples were then filtered through 20 pm cell strainers and cells were placed in T75 culture flasks in DMEM + Glutamax media (ThermoFisher, Cat: 10569010) enriched with 10% fetal bovine serum (ThermoFisher, Cat: 10082147) and 1% Antibiotic-Antimycotic (ThermoFisher, Cat: 15240062) at 37°C and 5% CO2. Cells were passaged when reaching >70% confluence with Trypsin- EDTA (ThermoFisher, Cat: 25200056), and were used at passages 2-5. For EP300 inhibition, cells were treated with 250 pg of I-CBP112 (EP300-INH) (Selleck Chem. Inc; Radnor. PA) for 48 hours, then harvested for downstream processing, including CUT&Tag. For SLIT2 in vitro experiments, dorsal fibroblast cultures were treated with either 250ug of SLIT recombinant protein (R&D, Cat no. aa 26-900) or PBS vehicle control for 48 hours, then used for downstream analyses.

[0225] CRISPR-Cas9 gene-editing of cultured fibroblasts

[0226] Predesigned TrueGuide™ Synthetic sgRNAs (SantaCruz Biotechnology) for mouse Robo2 knockout, Robo2 knock-in, Eid knockout. Eidl knockin. non-targeting guide RNAs for negative controls and TrueCut Cas9 Protein v2 (SantaCruz Biotechnology) were used for CRISPR-Cas9-mediated gene editing of >70% confluent primary' mouse dorsal and facial fibroblast cultures from GFP and B6 mice. The respective sgRNA and Cas9 were transfected into cultured fibroblasts using the Lipofectamine™ CRISPRMAX™ Cas9 Transfection Reagent (SantaCruz Biotechnology) per manufacturer’s protocol. Transfected cells were kept in culture for 48 hours before harvesting for injection or downstream analyses, including CUT&RUN. Gene editing efficiency for Robol- and Eidl- knockout fibroblasts was assessed via FACS using ROBO2 and EID1 antibodies, and via RT-qPCR using Robo2 and Eidl primers.

[0227] Wound treatment with SLIT2 and EP300 inhibitor

[0228] Recombinant SLIT2 (R&D, Cat no. aa 26-900) was injected into the base of dorsal excisional wounds at PODO at a concentration of 250ug / wound. I-CBP112 EP300 inhibitor (EP300-INH) (Selleck Chem, Inc; Radnor, PA) was injected into the base of dorsal excisional wounds at a concentration of 250 pg inhibitor / 30 pl PBS per wound. PBS was used as negative control for both SLIT2 and I-CBP112 wound injections at the same volumes as their respective treatment volumes.

[0229] Lentiviral Eidl Overexpression and Knockdown

[0230] Eidl overexpression and knockdown studies were conducted in CollalCre- ERT;Ai9 mice (fibroblasts express tdTomato). For Eidl overexpression, FLEXon Eidl:T2A:EGFP lentiviral vectors encoding a floxed mEidl.EGFP transgene with a proteolyzable T2A linker (control lentivirus contained EGFP alone; successfully transduced fibroblasts expressing transgene are EGFP+) were utilized. For Eidl knockdown, FLEXon EGFP:miR30-mEidl lentiviral vectors encoding a floxed miR30- based Eidl shRNA (control lentivirus contained scrambled miR30 sequence; successfully transduced fibroblasts expressing transgene are EGFP+) were utilized. Ultra-purified overexpression or knockdown Eidl lentivirus was delivered via four 15 pl intradermal injections at the wound site (>109 TU / ml). For the early transduction experiments, two applications were delivered at three days prior to wounding (POD -3) and two days post wounding (POD +2). For late transduction experiments, two applications were delivered at seven and ten days post wounding (POD 7 and 10).

[0231] Nuclei isolation from mouse dermis

[0232] For nuclei isolation, 100 mg of flash-frozen mouse dermis was finally minced with surgical scissors for 4 min. O.lx lysis buffer was used for a 2-minute incubation, followed by quenching with wash buffer and centrifugation at 4°C at 500 ref for 5 min. Following two wash and centrifugation steps, nuclei were resuspended in nuclei buffer (lOx Genomics), counted, and then processed using lOx Genomics’ Chromium platform. During nuclei retrieval, samples were filtered using 40pm (Flowmi) and 20pm filters (PuriSelect).

[0233] Single-cell ATAC sequencing

[0234] scATAC-seq was performed following lOx Genomics protocols. In brief, nuclei were isolated as described above from dermal mouse wounds. Mouse wounds from n = 6 mice were pooled for each condition (unwounded facial dermis, unwounded dorsal dermis, dl4 facial scars, dl4 facial scars treated with EP300 inhibitor, dl4 dorsal scars, and d!4 dorsal scars treated with EP300 inhibitor). Nuclei then underwent transposition, GEM generation and barcoding, post GEM incubation cleanup, library construction, and qualitative control on an Agilent Bioanalyzer High Sensitivity DNA chip (lOx genomics protocol CG000496 Rev B). Libraries were pooled and cDNA libraries sequenced on the Illumina platform.

[0235] scATAC-seq Data Processing and Analysis

[0236] lOx Genomics Cell Ranger tool cellranger-atac mkfastq was used to demultiplex raw base call (BCL) to fastq files. Using default parameters, cellranger-atac count was used to align sequencing files to the mouse genome (mm 10). Signac (v. 1.10.0) was then used to conduct downstream analysis. The following cutoffs were used: minimum 3000 peaks, maximum 30000 peaks, percent reads in peaks greater than 15%, blacklist ratio less than 0.05, nucleosome signal less than 4. and TSS enrichment score greater than 3. Following normalization, linear and non-linear dimensional reduction, nuclei were clustered using dimensions 2 to 30 and a gene activity matrix generated. Integration with scRNA-seq data w as performed using Signac’s “Integrating with scRNA- seq data” vignette.

[0237] Visium spatial analysis

[0238] Wound specimens were rapidly harvested and flash frozen in OCT. Using the Visium Tissue Optimization Slide and Reagent Kit, permeablization time was optimized at a thickness at 10 pm section and 37 minutes for mouse tissue. Following cryo-sectioning at -20 degrees onto gene expression slides the expression slide and reagent kit was used to produce sequencing libraries. NextSeq (Illumina) was used to sequence the libraries.Following demultiplication the raw FASTQ files and histology' images were processed by sample with the Space Ranger software for genome alignment. The raw space ranger output files for each sample was then read into a Seuratclass object in R using Seurat’s Load 1 Ox function. Data was normalized using the SCT transform with default parameters. To ascertain the integration of our scRNAseq and Visium spatial analysis we employed the FindTransferAnchors function from Seurat, which allowed the alignment of data using the two datasets. This cross platform linkage is performed serially in an unconstrained and constrained fashion.

[0239] CODEX spatial analysis

[0240] To spatially phenoty pe the mouse wound specimens, Co-Detection by Indexing (CODEX), a novel assay in which markers are labeled with oligonucleotide- conjugated antibodies and iteratively imaged between cyclic additions and washouts of dye-labeled oligonucleotides, was used. A custom CODEX panel was designed to assess wound cells within the tissue (Table 1). In brief, primary' antibodies were individually barcoded and validated using the commercial supplier's protocols. OCT for mouse or paraffin blocks for human xenograft (n = 3 per group) were sectioned at 8 pm thickness onto coverslips for CODEX antibody staining. Antigens were retrieved by standard citrate-EDTA processing prior to addition of CODEX antibodies. Using a CODEX integrated Keyence BZ-X instrument (Akoya Biosciences) image acquisition was then performed. Using software from Akoya Biosciences the raw images were processed, with cell segmentation, and rendering. 92 The CODEX was visualized using Akoya Biosciences Multiplex Analysis Viewer (MAV) in ImageJ. The resulting .fcs files were then concatenated in FlowJo and imported into the Monocle3 and STvEA R packages for further analysis. After debris removal, the processed UMAP manifold was analyzed through Monocle3 with a postmanifold threshold of >10,000 cells per cluster. Analysis of the protein staining patterns was then used to assign cell ty pes. The cell interactions were then inferred using STvEA for cell types at >2.5% of total abundance at k=20 nearest neighbors to quantify cell spatial interactions, and difierential interaction maps were generated using ggraph scores.

[0241] CUT&Tag

[0242] CUT&Tag experiments were performed on cultured fibroblasts treated with vehicle control or LCBP112 inhibitor as described in Kaya-Okur, et al., 202051using the CUT&Tag-IT Kit (Active Motif, Catalog No. 53160). Briefly, primary' dorsal and facial mouse fibroblast adherent cultures were dissociated with Accutase Cell Detachment Medium (Thermo Fisher Scientific) at 37 °C for 5 minutes and resuspended in PBS. Cellnumber was determined on hemocytometer and 100.000 cells per condition were used. Cells were bound to activated Concanavalin A beads permeabilized in 0.05% digitonin, incubated in Ipg per sample of histone H3K27ac primary antibody (Active Motif, Catalog No. 39034) at 4 °C overnight with orbital mixing (no primary antibody used for negative control). After antibody buffer washes, cells were incubated in 1: 100 dilution of guinea pig anti-rabbit secondary antibody at room temperature for 60 minutes. After antibody buffer washes, CUT&Tag-IT Assembled pA-Tn5 were added to reaction and tagmentation performed as described in the protocol. After stopping tagmentation reaction, DNA was isolated from supernatant and PCR amplification was performed using the provided i5 and i7 indexed primer pairs and run for 14 cycles, as described in the protocol. Double-sided size selection of the library was performed with SPRI beads (0.9X), and library was quantified using QuBit fluorometer and size distribution was visualized via TapeStation. Individual libraries were then pooled and a one-sided SPRI bead selection (0.9X) was performed on the pooled library. Sequencing was performed with NexSeq MID output (2 x 75bp PE) platform. Sequencing data was analyzed per Zheng Y et al (2020) (https: / / yezhengstat.github.io / CUTTag_tutorial). Briefly. adaptor sequences were trimmed using cutadapt and then aligned to mm39 mouse genome using Bowtie2. Spikein calibration and scale factor normalization was performed with E. coli genome carried over with pA-Tn5 and mapped to ASM584v2. Peak calling was performed with SEACR and peak annotation performed with ChIPseeker66 and differential H3K27ac analysis over merged peaks was performed with DESeq2.

[0243] CUT&RUN

[0244] CUT&RUN was performed on primary culture dorsal and facial control and Eici\ - knockout mouse fibroblasts per EpiCypher® CUTANA™ CUT&RUN protocol. Briefly, adherent cells were cross-linked in situ with 0.1% formaldehyde for 1 minute at room temperature, then detached with Accutase Cell Detachment Medium (Thermo Fisher Scientific) at 37 °C for 8-10 minutes and resuspended in PBS. Cell number w as determined on hemocytometer and 100,000 cells per condition were used. After binding to activated ConcanavalinA beads, cells were permeabilized in 0.005% digitonin containing antibody buffer and incubated with Ipg per sample of histone H3K27ac primary antibody (Active Motif, Catalog No. 39034) at 4 °C overnight with orbital mixing (Ipg per sample of rabbit IgG (EpiCypher SKU: 13-0042) antibody used for negative control). After XL digitonin buffer washes binding to pAG-MNase was performed, activated, and the reaction stopped with Stop Buffer Master Mix containing 0.027ng per sample of CUTANA™ E. coli Spike-in DNA (EpiCypher SKU: 18-1401). Cross-link was reversed overnight as described in the protocol and DNA fragments were isolated from supernatant with CUTANA™ DNA Purification Kit (EpiCypher SKU: 14-0050). Libraries were prepared with NEBNext® Ultra™ II DNA Library Prep Kit for Illumina (E7645S) and NEBNext® Multiplex Oligos for Illumina® and amplified as described in the protocol for 14 cycles. Double-sided size selection of the library was performed with SPRI beads (0.9X), and library was quantified using QuBit fluorometer and size distribution was visualized via TapeStation. Individual libraries were then pooled and a one-sided SPRI bead selection (0.9X) was performed on the pooled li brary. Pooled library' was sequenced on NovaSeq PEI 50 platform (Novogene) and sequencing analysis was performed as described under Methods in CUT&Tag.RESULTS

[0245] Embryonic origins determine fibroblasts’ intrinsic fibrogenic potential in cutaneous wound healing

[0246] To determine if the embryonic origins of fibroblasts affect scar formation, a four-site wounding model was developed in which there were created 2.5mm fullthickness excisional skin wounds on the face, scalp, ventrum, and dorsum of the mouse (FIG. 2A). Given that murine skin wounds contract due to the activity' of the subcutaneous panniculus camosus, wounds were stented using silicone rings to prevent rapid contraction and to maximize healing by secondary intention.11’26After re-epithelialization at postoperative day (POD) 14, wounds were harvested for histology and immunohistochemical (IHC) staining. Hematoxylin and eosin (H&E) and Masson’s trichome staining revealed that facial wounds had less scar formation, with significantly decreased dermal thickness and collagen deposition, especially when compared with dorsal wounds (FIG. 2B). As scar width is only one high-level w ay of assessing scar formation, we additionally performed immunohistochemical (IHC) staining of healed POD- 14 w ounds using antibodies for canonical fibroblast activity markers, including collagen ty pe I (COL1), alpha smooth muscle actin (aSMA), and collagen type III (COL3). Imaging and quantification of IHC data showed that these fibrosis markers were significantly' increased in intensity in dorsal wounds compared to facial wounds, consistent with increased scar formation (FIG. 2C).

[0247] A delay in re-epithelialization itself can prolong inflammation (e.g., in chronic wounds) and lead to increased fibrotic response, 28 which could explain the decreased scarring phenotype observed in faster-healing facial and scalp wounds compared to ventral and dorsal wounds. However, although facial and scalp wounds healed at similar rates (fullre-epithelialization by POD 11) (FIGS. 7A-7E), facial wounds nevertheless healed with significantly decreased scarring compared to scalp wounds (FIG. 2B). These results indicated that varying rates of wound closure may not fully explain anatomically variant scar formation. To assess whether the fibrotic properties of the skin are specific to their dermal embry onic origins, two distinct transplantation experiments were performed. First, full-thickness skin grafts from the face, scalp, ventrum, and dorsum of RFP+ mice were harvested and transplanted onto the dorsum of wildtype (C57BL / 6J) mice (FIG. 3A). Following donor skin engraftment (PODIO), incisional wounds were made on the RFP+ skin grafts. After 14 days of healing, IHC staining showed significantly reduced COL1 deposition in the wounds of facial skin engrafted onto the dorsum compared to the wounds of dorsal, scalp, or ventral skin engrafted onto the same site (FIGS. 3B and 3C). These results showed that the native skin microenvironment, rather than the anatomical location per se, dictates the degree of wound fibrosis. Second, to determine whether such wound healing differences may be influenced by fibroblasts specifically, fluorescence-activated cell sorting (FACS) was used to isolate facial, scalp, ventral, and dorsal fibroblasts from a GFP+ mouse using a previously published lineage-negative gating.10and intradermally injected each population separately into the dorsum of wildly pe mice (FIG. 4A). Compared to scalp, ventral, and dorsal fibroblasts, facial fibroblasts showed the lowest COL1 deposition by IHC co- localization with GFP (FIGS. 4B and 4C). Together, these experiments demonstrated that fibroblasts of different embryonic origins display diverse fibrogenic potentials and retain their origin-specific cell-intrinsic fibrogenic potential even when transplanted onto a distant location.

[0248] Robo2 expression defines a subpopulation of cranial neural-crest-derived fibroblasts with reduced fibrogenic potential

[0249] To investigate the mechanisms underlying the disparate fibrogenic potential of fibroblasts, differences in transcriptional activity were next assessed. Using the ‘four- site wounding model,’ we performed single-cell RNA sequencing (scRNA-seq) of FACS- isolated fibroblasts (via lineage-negative gatinglO) from those four anatomical sites at POD 7 and POD 14 (FIG. 5 A). Fibroblasts from unwounded skin at the four sites were also evaluated as controls. What was found was distinct clusters of fibroblasts in unwounded skin, at POD 7, and at POD 14 containing a mix of cells from all four sites, with six transcriptionally distinct fibroblast clusters identified at POD 14 (FIG. 5B). Interestingly, facial fibroblasts harvested from wounds at POD 14 predominantly grouped into clusters 0, 2, and 4 (FIG. 5C, left). Pathway analysis showed that clusters 0, 2, and 4demonstrated neural crest-like features, with prominent expression of genes involved in TGF- and WNT signaling pathways (FIG. 5D). Furthermore, using CytoTRACE27to infer the cellular differentiation states based on sequencing data, it was found that clusters 2 and 4 had the lowest predicted differentiation states (FIG. 5C, right). Given these results, it was concluded that clusters 2, 4, and to a lesser extent cluster 0 may possibly contain more progenitor-like fibroblast populations.

[0250] To identify fibroblast subpopulations responsible for wound repair with reduced fibrosis, next looked for were genes encoding cell surface proteins that were both highly expressed in facial fibroblasts from clusters 0, 2, or 4 and broadly involved in neural crest and / or regeneration. The analysis revealed four genes that fit those criteria - Robo2. CdM, Ncami, and Gpc3 (FIG. 5E). Pseudotime trajectory analysis supported that these genes were highly expressed in these clusters (FIG. 5F). FACS analysis of fibroblasts isolated from the four anatomical regions at the time of wound closure (POD 14) showed enrichment of all these cell surface proteins on facial fibroblasts compared to fibroblasts from the other sites (FIG. 5G). In particular, ROBO2+cells were the predominant population in facial fibroblasts (p < 0.05) (FIG. 5G). Collectively, these results suggest that Robo2 expression may define a subpopulation of facial fibroblasts with reduced fibrotic potential within the cranial neural-crest derived dermis.

[0251] ROBO2 in facial fibroblasts promotes wound healing with reduced fibrosis

[0252] Since the greatest differences in scarring were observed between facial and dorsal wounds (FIGS. 2B-2C and FIGS. 3A-3B and 4A-4C), additional transplantation experiments were performed to gain insight into the role of ROBO2+ fibroblasts in fibrosis during wound healing. Facial fibroblasts from GFP mice were harvested, sorted into ROBO2+or ROBO2" populations by FACS, and each population (100,000 cells / wound) injected into the periphery of dorsal wounds on postnatal-day-60 wildtype mice (FIGS. 6A, 7A). Dorsal wounds injected with ROBO2+or ROBO2" fibroblasts were re- epithelialized by POD 14, at which time wounds were harvested and compared with untreated facial POD 14 wounds (FIG. 6 A). We confirmed engraftment of transplanted fibroblasts by FACS, which showed that 8% and 18% of fibroblasts were GFP+ in the ROBO2- and ROBO2+ fibroblast-injected wounds, respectively, at POD 14 (FIGS. 6D and 10B). ROBO2+fibroblast-treated dorsal wounds displayed decreased dermal thickness and collagen deposition compared to ROBO2 fibroblast- treated wounds on H&E and Masson's trichrome staining (FIG. 6B). Furthermore, ROBO2+fibroblast-treateddorsal wounds healed similarly as untreated facial wounds, with reduced fibrosis and increased hair follicle regeneration (FIG. 6B). The appearance of hair follicles in ROBO2+ fibroblast-treated dorsal wounds but not in untreated facial wounds (containing a mixture of ROBO2-low and ROBO2-high fibroblasts) indicates that a supraphysiological enrichment of ROBO2 signaling may be required for hair follicle regeneration not observed in native facial wound healing. However, it is also possible that additional pro- regenerative factors in ROBO2+ fibroblasts contribute to the increased regenerative outcome. Picrosirius red staining and image processing with Uniform Manifold Approximation and Projection (UMAP) visualization of extracellular matrix (ECM) ultrastructure revealed that ROBO2+fibroblast-treated wounds had ECM features overlapping with those of untreated facial wounds (FIGS. 6C and 28A). Moreover, their ECM were entirely distinct from the fibrotic ECM of ROBO2- fibroblast-treated dorsal wounds (FIG. 6C, 28A). Immunostaining demonstrated that SOXIO, a marker for neural crest cells and neural crest cell derivatives,28was upregulated in ROBO2+fibroblast- treated dorsal wounds compared to ROBO2 fibroblast-treated dorsal wounds, which also showed increased COL1 staining (FIGS. 6D and 28B). In addition, ROBO2+ fibroblast- treated wounds exhibited lower expression of fibrosis markers Co / 1, Co / 3, and aS ma than ROBO2" fibroblast-treated wounds (FIGS. 7B and 28C). Further supporting the reduced fibrotic phenotype of ROBO2+fibroblasts, ELISA of wound ly sates found lower COL1 levels in ROBO2 fibroblast-treated than in ROBO2" fibroblast-treated wounds (FIGS. 7C and 28D). Collectively, these results show that ROBO2 fibroblasts are less fibrogenic than ROBO2' fibroblasts, and when injected into dorsal ounds, are sufficient to promote healing with reduced fibrosis at levels similar to that in native facial wounds.

[0253] To determine if RoboZ expression itself is responsible for reduced scarring, CRISPR-Cas9-mediated Robo2 knockout (KO) and overexpression (OE) was performed in facial fibroblasts from GFP+ mice; then these cells were separately injected and CRISPR-control cells (with scrambled guide RNA) into the periphery of dorsal w ounds of wildtype mice (100,000 cells / wound) (FIG. 6E). At POD 14, a comparison was made of wound healing in these treated dorsal wounds to those of untreated facial and untreated dorsal wounds on other wildtype mice (FIG. 6E). FACS protein level assessment and RT- qPCR gene expression quantification confirmed 80% RoboZ KO and 75% Robo2 OE efficiencies (10-fold Robo2 KO and 1.5 to 2-fold Robo2 OE) by CRISPR-Cas9 compared to their respective KO and OE controls (FIGS. 23A and 23B). Compared to negative control fibroblast-treated dorsal wounds, untreated facial wounds, and untreated dorsalwounds, Robol KO fibroblast-treated dorsal wounds healed with significantly greater dermal thickness and collagen deposition, more fibrotic ECM ultrastructure, decreased SOXIO staining, and increased COL1 staining in the dermis (FIGS. 6F-H and 11C). Compared to untreated and CRISPR OE control fibroblast-treated dorsal wounds, Robo2 OE fibroblast-treated dorsal wounds healed with significantly reduced scar width, increased hair follicle number, less fibrotic ECM ultrastructure, increased SOXIO staining, and decreased COL1 staining in the dermis (FIGS. 6F-H. 11C). Furthermore, expression of fibrosis markers and COL1 levels in wound lysate were significantly increased in Robol KO fibroblast-treated dorsal wounds compared to all other groups (FIGS. 7D, E). Conversely, Robol OE fibroblast-treated dorsal wounds healed with decreased dermal thickness and collagen deposition, less fibrotic ECM ultrastructure, increased SOXIO dermal staining, decreased COL1 dermal staining, decreased fibrosis marker expression, and decreased wound supernatant COL1 levels when compared with negative control fibroblast-treated dorsal wounds and untreated dorsal wounds (FIGS. 6F-H, 7D, 7E, 29A, 29B). Notably, transplantation of Robo2 OE fibroblasts enabled dorsal wounds to heal with all the attenuated fibrotic features observed in untreated facial wounds (Fig. 6F-H, 7D, 7E, 11C, 29A, 29B). Although Robol KO fibroblast-treated dorsal wounds healed with similar scar width and lack of hair follicles compared to untreated and CRISPR KO control fibroblast-treated dorsal wounds (FIG. 6F), dorsal wounds injected with / o / w2- KO fibroblasts exhibited more fibrotic ECM ultrastructure, increased COL1 dermal staining, and increased fibrosis marker expression when compared with KO control fibroblast-treated dorsal wounds (FIGS. 6G, 6H, 11C, 29A). Whereas injection of CRISPR-control facial fibroblasts led to some reduced fibrotic features compared to untreated dorsal wounds (FIG. 6G, 6H. Fig. 29A), Robol OE and KO respectively- augmented and countervailed the antifibrotic effects of ectopic facial fibroblasts (FIGS. 6F-H, and 29A and B), demonstrating that transplantation of fibroblasts with disrupted fibrotic potential underpinned major changes in scarring phenotype. Mere injection of fibroblasts per se was not sufficient to drastically alter fibrosis (FIG. 6F). Together, these CRISPR- Cas9-mediated KO and OE experiments demonstrated that Robo2 is both necessary and sufficient to promote dorsal wound healing with reduced fibrosis (FIG. 61), akin to that of facial skin.

[0254] Lastly, as SLIT2 is a known ligand of the ROBO2 receptor,29SLIT2 recombinant protein was used to examine the upstream regulation of ROBO2 signaling (FIGS. 1). Mouse dorsal fibroblast cultures were first treated with either SLIT2 or vehiclecontrol and compared their fibrotic activity to that of untreated mouse facial fibroblast cultures. Treating dorsal fibroblasts with SLIT2 increased their ROBO2 and SOXIO levels as detected by IHC and FACS, respectively (FIGS. 8A, 8B, 13B, 13C). Furthermore, SLIT- treated dorsal fibroblasts showed a reduction in the gene expression and protein levels of several fibrosis markers, including SMA, COL1, and TGF-betal, mirroring the phenotype of untreated facial fibroblasts (FIGS. 8A, 8C, 8D, 13C, 30A, 30B). To test the role of SLIT2 in vivo, SLIT2 was applied to dorsal wounds via local injection at the time of wounding to evaluate whether increasing SLIT2 levels in wounds alone could mimic facial healing with reduced fibrosis (FIGS. 9A, 14A). Consistent with the findings in wounds transplanted with ROBO2+fibroblasts, SLIT2 -treated dorsal wounds showed a reduced scarring phenotype akin to that seen in untreated facial wounds as assessed by histology, ECM ultrastructure analysis, IHC, fibrosis marker expression, and COL1 levels in the wound ly sate at POD 14 (FIGS. 9B-F, 14B, 14C, 31A-31C). SLIT2 increased ROBO2 levels in both cultured dorsal fibroblasts (FIGS. 13B, 13C) and dorsal wounds (FIG. 31A), thus promoting a positive ligand-receptor feedback loop that enhances downstream signaling. These results demonstrated that SLIT2. an upstream ligand of ROBO2, is also important in inhibiting fibrosis during wound healing (FIGS. 9G and 3 ID).

[0255] EID1 mediates the reduced fibrotic potential of ROBO2+ facial fibroblasts

[0256] To understand how ROBO2+facial fibroblasts promote wound repair with less fibrosis at a gene regulatory level, the scRNA-seq data was examined for facial fibroblast- enriched genes encoding transcriptional regulators. Four such genes - Eidl , Trspl, Tcfl / lZ, and GUI - were highly expressed in facial fibroblasts and in the previously identified fibroblast cluster 2 (FIGS. 10A, 15 A). As Eidl has been shown to inhibit cellular differentiation by mediating transcriptional repression,30’31it was interrogated. Immunostaining confirmed that there was a significantly higher proportion of cells with EID1 and ROBO2 co-localization in facial wounds than in dorsalwounds at POD 14 (FIG. 10B and 15B).

[0257] To determine whether EID1 acts downstream of ROBO2, CRISPR- Cas9 was used to suppress the expression of either RoboZ or Eidl in mouse facial fibroblast cell lines and quantified the reciprocal effects on their protein levels via IHC and FACS (FIGS. HA and 16A). Both IHC and FACS showed that EID1 levels were decreased in RoboZ KO fibroblasts while ROBO2 levels were unchanged in Eidl KO fibroblasts when compared with those in control facial fibroblasts (FIG. 11B, 11C, 16B, 16C). Furthermore, ROBO2 activation by SLIT2 treatment significantly increased EID1 transcript and proteinlevels in facial fibroblasts compared to control PBS (FIG. S16D). This helped confirm that Robol regulates the downstream expression of Eidl. Consistent with the anti- fibrogenic roles of Robol and Eid\ . suppression of either gene also increased collagen I production as assessed by IHC (FIGS. 11B, 16B). EID1 represses transcription by binding to and inhibiting the activity7of EP300,30,31a histone acety ltransferase.32Increased numbers of EP300+ fibroblasts were additionally observed in both Robol and Eidl KO cell lines compared to control facial fibroblasts (FIGS. 11B. 11C. 16B. 16C), demonstrating that EP300 synthesis is also negatively regulated by ROBO2 and EID1. EP300 level is also suppressed by ROBO2 and EIDE ROBO2- and EIDl-mediated suppression of EP300 occurs in part via reduction of Ep300 mRNA levels, as evident in the negative regulation of the amount of Ep300 transcripts by ROBO2 and EID1 in the CRISPR cell lines (FIG. 16E). In sum, these data helped delineate the epistatic hierarchy of the ROBO2-EID1- EP300 signaling axis.

[0258] To test the role of EID 1 in wound healing, CRISPR-Cas9-generated Eidl KO, Eidl OE, and negative (scrambled guide RNA) control facial fibroblast cell lines from GFP+mice were used and separately injected them (100,000 cells / wound) into the periphery7of dorsal wounds of wildtype P60 mice (FIG. 12A). Fibroblast-treated dorsal wound healing outcomes were compared with those of untreated facial and untreated dorsal wounds on wildtype mice (FIG. 12A). The efficacy of / A / l KO was confirmed by the decreased levels of EID1 and SOX10 and increased levels of COL1 in cultured fibroblasts, as assessed by IHC and FACS (FIGS. 13A, 13B). See also FIGS. 18A and 18B showing a 5 to 10-fold decrease and 1.2 to 2-fold increase in EID1 levels in cultured facial fibroblasts compared to their respective control cell lines, as assessed by RT-qPCR and ICC. Eidl KO fibroblast-treated dorsal wounds healed similarly as untreated dorsal wounds, with significantly greater dermal thickness and collagen deposition, more fibrotic ECM ultrastructure, decreased dermal SOX10 staining, increased dermal COL1 staining, increased fibrosis marker expression, and increased COL1 levels in the wound lysate when compared to negative control fibroblast-treated dorsal wounds and untreated facial wounds (FIGS. 12B-12D. 13C, 13D, 32A-32C). Conversely, Eidl OE fibroblast-treated dorsal wounds healed with decreased dermal thickness and collagen deposition, less fibrotic ECM ultrastructure, increased dermal SOX10 staining, decreased dermal COL1 staining, decreased fibrosis marker expression, and decreased wound supernatant COL1 levels when compared with negative control fibroblast-treated and untreated dorsal wounds (FIGS. 12B-12D, 13C, 13D, 32A-32C). In line with ROBO2-enrichment indorsal wounds, the additional CRISPR-mediated Eidl upregulation in the already ROBO2 / EID1 -enriched facial fibroblasts may partially account for hair follicle regeneration in treated dorsal wounds but not in untreated facial wounds, although other pro-regenerative factors in Eid 7-OE fibroblasts may also be at play. Conversely, although Eidl KO fibroblast-treated dorsal wounds healed with similar scar width and lack of hair follicles as untreated and KO control fibroblast-treated dorsal wounds, compared to injection of KO-control fibroblasts the addition of Eidl- O fibroblasts within dorsal wounds led to more fibrotic ECM ultrastructure, increased fibrosis marker expression, and increased wound lysate COL1 levels (FIGS. 12B-12C, 32A-32C). Notably, like Robol OE fibroblast-treated dorsal wounds (FIG. 6F-6H, 7D, 7E), Eidl OE fibroblast-treated dorsal wounds also healed with all the attenuated fibrotic features observed in untreated facial wounds (FIGS. 12B-12D, 13C, 13D). Together, these experiments revealed that fibroblast expression of Eidl downstream of Robol is also necessary and sufficient to decrease fibrosis in dorsal wounds and promote more facial-like healing characteristics (FIG. 12E).

[0259] Given the importance of EID1 in wound repair with reduced fibrosis, it was sought to further test the effect of Eidl knockdown (KD) and OE in fibroblasts in vivo. FLEXon eGfp:miR30-mEidl lentiviral vectors encoding floxed miR30- based Eidl shRNA were developed, that when injected into the dorsum of Collal-CreERT;Ai9 mice, enabled fibroblast-specific Eidl KD upon tamoxifen treatment. For control treatment (Eid 1 -KD control), we used the FLEXon eGfp: scramble miR30 lentivirus. Conversely, dorsal injection of FLEXon Eidl:T2A.eGfp lentiviral vectors encoding a floxed mEidF. eGfp transgene with a proteolyzable T2A linker enabled fibroblast- specific Eidl OE upon tamoxifen treatment. In this case, the FLEXon eGfp lentivirus was used as control (Eidl-O control). The effects of Eidl KD or OE were tested during early stages of wound healing by injecting lentiviruses 3 days before and 2 days after wounding (POD -3 / +2). Similarly, lentiviruses were injected 7 and 10 days after wounding (POD7 / 10) to test the effects of Eidl manipulation during later stages of wound healing. Using this strategy, we observed close to one-hundred percent viral transduction efficiency in wound fibroblasts, leading to an overall two-fold increase and a three-fold decrease in EID1 levels by the overexpression and knockdown constructs, respectively (FIG. 19A). At early stages of repair (POD -3 / +2) characterized by inflammation, granulation, and early ECM synthesis, Eidl OE in dorsal wounds led to reduced scarring with regeneration of hair follicles, less fibrotic ECM architecture, and increased dermal levels of SOX 10 compared to control(Eidl-OEC) treatment (FIGS. 14A-14C, 19B-19D). In contrast, Eidl KD in dorsal wounds had no impact in the fibrotic phenotype compared to control (Eidl-KDC) (FIGS. 14A-14C, 19B-19D). Similar effects in dorsal wound healing were observed when Eidl OE and KD were performed at later wound healing stages (POD 7 / 10), when processes like fibroblast migration, ECM deposition, and remodeling predominate (FIGS. 15A-15C, 20A-20C). Collectively, these findings further demonstrated that Eidl expression in fibroblasts is sufficient for wound healing with reduced fibrosis.

[0260] EP300 inhibition downstream of EID1 in dorsal fibroblasts promotes facial-like wound healing with reduced fibrosis

[0261] EID 1 -mediated EP300 inhibition prevents the dilferentiation of neural crest- derived cells, thus maintaining them in a more progenitor-like state.33-35Bromodomain inhibitors have been developed that likewise suppress EP300 activity, and these small molecules have emerged as a promising class of anticancer therapeutics.36,37In light of this, the ability of I-CBP112, a commercially available small- molecule EP300 inhibitor, to mitigate scar formation in dorsal skin wounds was investigated.

[0262] Mouse dorsal skin fibroblasts were cultured in vitro with 250 g of I-CBP112 (EP300i) (Selleck Chem, Inc; Radnor, PA)38and compared to cultured dorsal and facial fibroblasts treated with vehicle control (FIGS. 16 A, 21 A). Immunostaining showed that EP300 inhibition decreased EP300 levels in cultured dorsal fibroblasts to levels similar to those in cultured facial fibroblasts (FIGS. 16B, 21B). Compared to vehicle control. I- CBP112 also decreased the amount of Ep300 transcripts in primary cultures of dorsal fibroblasts (FIG. 21 C). Both transcript and protein levels of Collagen type I in cultured dorsal fibroblasts were also significantly reduced with I- CBP112 treatment (FIGS. 16C, 16D, 21D, and 33A). Furthermore. EP300 inhibition also increased the proportion of dorsal fibroblasts expressing SoxlO, as assessed by FACS, to a level similar to that observed in facial fibroblasts (FIGS. 16E and 33B). These data show that bromodomain inhibitors like I-CBP112 may decrease the fibrotic activity of dorsal fibroblasts and promote a more facial fibroblast-like phenotype.

[0263] Given the promising in vitro results. EP300 suppression in vivo was next evaluated. Splinted excisional dorsal wounds in wildtype mice were injected with 250 pg EP300i or PBS control at the time of wounding. EP300i treatment did not significantly alter the rate of wound re-epithelialization compared to PBS control treatment, as both groups healed by POD 14 (FIG. 22A). Wounds were harvested at POD 14 for analysis and compared with untreated facial wounds (FIG. 17 A). EP300i-treated dorsal wounds healedsimilarly as untreated facial wounds, with significantly decreased dermal thickness and collagen deposition, less fibrotic ECM ultrastructure, and decreased dermal COL1 staining (FIGS. 17B, 22B, 22C). Furthermore, increased dermal SOXIO staining, lower expression of fibrosis markers, and significantly lower COL1 levels in the wound lysate for EP300i- treated dorsal wounds were noted when compared with PBS-treated dorsal wounds (FIGS. 17B, 16F-16I, 22C, 34A, 34B). Because SoxlO is expressed in not just neural crest cells but also their derivatives (glia, melanocytes) in adult skin,28SOX9, an early neural crest cell specification marker,39was stained for to additionally assess for neural-crest-like identity7in wounds. At POD 14, SOX9 levels in EP300i-treated dorsal wounds were similar to those observed in untreated facial wounds, and virtually absent in PBS-treated dorsal wounds (FIG. 16G, bottom panel, 22C, bottom panel). Moreover, to trace the lineages of SOX9+ and SOX10+ cells following dorsal wounding, tamoxifen- induced Sox9- CreERT;mTmG and SoxlO-CreERT;mTmG mice were generated and similar wounding experiments were performed with or without EP300i (FIGS. 18A, 18B left, 24A, 24B left). EP300i -treated dorsal wounds demonstrated an increase in the number of GFP+SOX9+and SOXlCf cells following wounding compared to PBS-treated dorsal wounds, similar to those observed in untreated facial wounds (FIGS. 18 A, 18B right, 24A, 24B right). These results corroborate the fibroblast-specific increase in SOXIO levels noted in cultured and FACS-isolated dorsal fibroblasts treated with SLIT2 protein (FIGS. 8A, 8B, 13C, 13D) or EP300i (FIGS. 16E, 33B). In all. these data further demonstrated that EP300 inhibition in dorsal wounds can phenocopy facial wound healing with reduced scarring and increased neural crest cell- like features.

[0264] Given the potential off-target effects of the EP300 inhibitor, similar wounding experiments were performed in transgenic mice deficient in A / GOO. Because homozygous null Ep300 mutations lead to embryonic lethality,40Col lai- CreERT;mTmG:Ep300+ / - mice were generated with fibroblast-specific Ep300 KO upon tamoxifen treatment, and their dorsal wound healing was compared to that of PBS- and EP300i- treated dorsal wounds, as well as unwounded dorsal skin, of control mice (Called- CreERT;mTmG) (FIGS. 19A, 25 A). Compared to dorsal wounds on control mice, dorsal wounds on Ep300 KO mice healed with reduced scarring and regeneration of hair follicles, less fibrotic ECM architecture, and upregulation of dermal SOXIO, recapitulating the phenotypes seen in EP300i-treated wounds and unwounded dorsal skin (FIGS. 19B-19D, 25B). Compared to dorsal wounds on control mice, dorsal wounds on Ep300 KO mice healed with reduced scarring and presence of hair follicles, less fibrotic ECM architecture, and upregulation ofdermal SOXIO, recapitulating the phenotypes seen in EP300i-treated wounds (FIGS. 35A-C). Together, these data suggest that disrupting EP300, including with just a single dose of a small molecule inhibitor, may induce a typically scar-prone site like the dorsal skin to heal more like cranial neural crest-derived facial skin with reduced scarring (FIG. 17C).

[0265] Given the use of EP300i has the potential to translate to the clinical bedside, the facial skin-like wound healing of EP300i-treated dorsal wounds was further corroborated with scRNA-seq experiments. For scRNA-seq, facial and dorsal unwounded skin was harvested along with facial and dorsal wounds 14-days after PBS or EP300i treatment at the time of wounding (FIG. 17D). Cells within unwounded skin and wound samples segregated into 32 clusters based on gene expression profiles (FIG. 20 A. or 33 in FIG. 36A), and these were further annotated with cell type identities based on differentially expressed genes in each cluster (FIGS. 20B, 36B). Across different samples, the fibroblast cluster exhibited significant changes in distribution and was thus isolated for additional analysis (FIGS. 20C, 36C).

[0266] Fibroblasts across all samples segregated into 9 different clusters based on gene expression profiles (FIG. 17E). Fibroblast clusters 1-4 were the most over- represented subtypes in facial wounds compared to PBS-treated dorsal wounds (FIG. 17E, 17F). Among these, fibroblast clusters 1-3 prominently featured genes associated ribosomes and ribosomal function (FIG. 17G). In light of the importance of ribosomal biogenesis to proper neural crest cell function,41clusters 1-3 may thus represent neural-crest cell-like fibroblast subsets that harbor reduced fibrotic activity7. In contrast, cluster 6 was the most predominant in the PBS-treated dorsal wounds relative to facial wounds (FIGS. 17E, 17F), and may represent a pro-fibrotic fibroblast subpopulation. In fact, pathway analysis of genes enriched in cluster 6 implicated many in mechanotransduction and ECM synthesis (FIG. 17G), nominating cluster 6 as the '‘mechanoresponsive” fibroblasts associated with fibrosis in multiple contexts, including scarring and tumorigenesis.26,42,43EP300 inhibition in dorsal wounds dramatically decreased the number of cluster 6 fibroblasts (while increasing the numbers of cluster 1, 2, 3. 4, 5. and 7 fibroblasts), therefore shifting the fibroblast population profiles towards those of less fibrotic PBS- or EP300-treated facial wounds (FIGS. 17E, 17F). The attenuation of cluster 6 profibrotic fibroblasts in dorsal wounds by EP300 inhibition is consistent with the decreased scarring phenoty pe observed in the in vivo EP300i-treated stented excisional dorsal wounds (FIG. 17B). These results are notable as it has been demonstrated in other studies that suppressing mechanosensitivefibroblastpopulations through inhibition of cellular components of mechanotransduction (e.g., FAK. YAP. PIEZO) prevents or abrogates scar formation.11,43’44This study shows that EP300 may be yet another target for anti- fibrotic therapies.

[0267] To ascertain the transcriptional overlap between the FACS-isolated fibroblasts (FIGS. 2A-2G) and fibroblasts from the EP300i experiment (FIG. 17E), Seurat anchor label transfer was performed (FIGS. 20D left, 36D left). Facially enriched FACS-isolated fibroblast clusters 0, 2. and 4 (FIG. 5C, left) were highly represented in fibroblast clusters 1-5 and 7-9 but not in cluster 6 (mechanoresponsive) from the EP300i experiment (FIG. 20D right, FIG. 36D right), providing further evidence for the regenerative phenotype of fibroblast clusters 1-3 and the profibrotic phenotype of fibroblast cluster 6 from the EP300i experiment.

[0268] Complementing the scRNA-seq results, parallel single-cell assay for transposase-accessible chromatin with sequencing (scATAC-seq) revealed that cells from facial and dorsal unwounded skin and facial and dorsal wounds 14-days after PBS- or EP300i -treatment segregated into 32 clusters based on their chromatin profiles (FIGS. 21A. 21B, 37A, 37B). Integration of scATAC-seq data with our prior scRNA-seq results helped annotate these scATAC-seq clusters into distinct cell types (FIGS. 21C, 21D, 37C, 37D). Focus was made on the fibroblasts from the scATAC-seq data, and 12 clusters were identified within the fibroblast population based on their open chromatin profiles across experimental groups (FIGS. 22A, 22B, 38A. 38B). Notably, scATAC-seq fibroblast clusters 3 and 5 were expanded in dorsal PBS-treated wounds relative to facial PBS-treated wounds (FIGS. 22B, 22C, 38B, 38C). Conversely, scATAC-seq clusters 0 and 4 fibroblast populations were expanded in facial wounds relative to dorsal wounds (FIGS. 22B, 22C, 38B, 38C). Gene Ontology analysis showed that while dorsal wound-predominant clusters 3 and 5 were respectively enriched for genes involved in Wnt signaling and collagen fiber formation, facial wound-predominant clusters 0 and 4 were respectively associated with processes like epidermis development and blood vessel endothelial migration (FIG. 22D, 38D). In addition to promoting wound healing and hair follicle neogenesis, the pleotropic Wnt pathway is also known to increase collagen synthesis and fibrosis,1’45’47and its association with dorsal fibroblast subclusters is consistent with an increased scarring phenotype. On the other hand, the formation of the epidermis and vasculature is critical for regenerative healing and is in line with the reduced fibrotic phenotype of facial fibroblasts. Overall, the chromatin accessibility states between facial and dorsal wounds are distinct and readily distinguishable by the scATAC-seq profiles of fibroblasts (FIG.22C, top-middle and -right vs. bottom-middle and -right / FIG. 38C, top-middle and -right vs. bottom-middle and -right), supporting their disparate developmental origins and cell- intrinsic fibrogenic potential.

[0269] Spatial analysis identifies and defines regenerative facial fibroblasts

[0270] To further explore the significance of EP300 inhibition in wound repair, the lOx Genomics Visium platform was applied to analyze gene expression in the context of the spatial environment on wounds at POD 14 (FIG. 23 A). Anchor-based integration of the scRNA-seq fibroblast data and the spatial transcriptomics data was performed to assign fibroblast cluster identity' probabilities to each tissue coordinate and examined the spatial distribution of fibroblast subsets within each sample (FIG. 23B). Through this analysis, it was found that scRNA-seq-defined fibroblast cluster 3 (neural crest cell-like) was enriched in facial wounds compared to dorsal wounds (FIG. 23C, FW PBS vs. DW PBS). In contrast, scRNA-seq-defined fibroblast cluster 6 (mechanoresponsive) was enriched in dorsal wounds compared to facial wounds (FIG. 23C, DW PBS vs. FW PBS). Furthermore. EP300i treatment led to an attenuation of fibroblast cluster 6 identity and a slight induction of fibroblast cluster 3 identity within healed dorsal wounds at POD 14, when compared with PBS treatment (FIG. 23C, DW EP300i vs. DW PBS). In line with scRNA-seq results (FIG. 17F), EP300i treatment led to an attenuation of the spatial transcriptomic signature of fibroblast cluster 6 and a slight induction of the spatial transcriptomic signature of fibroblast cluster 3 within healed dorsal wounds at POD 14, when compared with PBS treatment (FIG. 23C, DW EP300i vs. DW PBS).

[0271] To assess putative communication networks among cell types in facial and dorsal wounds, differential interaction maps were generated comparing the groups based on cell-cell spatial co-localization. Dorsal wounds showed overall reduced interactions compared to facial wounds (FIG. 23D, left), which had greater interactions between fibroblast, monocytes, and epithelial cells. Following EP300i treatment, dorsal wounds showed greater cell-cell interactions compared to PBS-treated dorsal wounds (FIG. 23D, right). The specific cell-cell interactions in EP300i-treated dorsal wounds resembled those prominent in facial wounds, suggesting that EP300i reorganized the cellular interactome in dorsal wounds to mirror that observed in facial wounds healing with reduced fibrosis (FIG. 23D). Collectively, these data suggested that similar RNA-defined fibroblast subclusters existed in situ at a spatial level and contributed to the regenerative phenotype of facial wounds.

[0272] Spatial phenotyping by CODEX demonstrates that EID1+ROBO2+ fibroblasts exist in wounds that heal with reduced fibrosis

[0273] Building upon the spatial transcriptomic analyses described above, CO- Detection by indEXing (CODEX) was used to further spatially phenotype at the protein level facial and dorsal wounds treated with or without EP300i at POD 14 (FIG. 23A). For CODEX, a panel of 41 protein markers of different cell types was selected within the dermis (Table 1).Table X. CODEX protein panel. List of protein markers with their associated barcodes forCODEX experiments performed in this study.

[0274] These protein markers were sequentially labeled with and iteratively imaged via cyclic additions and washouts of dye-labelled oligonucleotide-conjugated antibodies.48Across all treatment groups. 15 cell clusters were defined based on CODEX protein signatures (FIGS. 23E, 24A, 24B, 39A).

[0275] Five CODEX-defined fibroblast clusters were identified based on protein expression levels, and the proportions of such fibroblast clusters varied among the treatment groups (FIG. 23F). CODEX fibroblast cluster 3 was most specifically enriched in PBS- treated dorsal wounds compared to facial wounds and unwounded skin, and was drasticallydiminished in EP300i-treated dorsal wounds (FIG. 23F, boxed). On the other hand, CODEX fibroblast cluster 4 was most enriched in EP300i-treated dorsal wounds compared with other groups (FIG. 23F, boxed). Moreover, assessment of combined levels of SLIT2, R0B02, and EID1 in each CODEX fibroblast cluster revealed that cluster 3 had the lowest levels and cluster 4 had the highest levels of these proteins (FIG. 6G, boxed), further suggesting that the former may represent a profibrotic fibroblast subtype and the latter may harbor a neural - crest-like pro- regenerative phenotype. Consistent with this. CODEX fibroblast cluster 3 displayed higher levels of many ECM proteins and fibroblast markers, including the myofibroblast protein aSMA, compared to CODEX fibroblast cluster 4 (FIG. 39B).

[0276] Differential cell-cell interaction scores were then calculated from CODEX spatial data based on K-nearest-neighbour localization. Compared to other experimental groups, PBS-treated dorsal wounds showed the greatest interactions between CODEX fibroblast cluster 3 and CD8 T-cells (FIG. 24B). In contrast, EP300i- treated wounds showed the greatest interactions between CODEX fibroblast cluster 4 and epithelial cells (FIG. 24B). These findings were consistent with more robust inflammation and re-epithelialization that occur in scarring and regenerative wounds, respectively.

[0277] Finally, examining the combined expression levels of Robo2 and Eidl in fibroblast clusters from scRNA-seq data (Fig. 17E) revealed that scRNA-seq-defined fibroblast clusters 5, 7, and to a lesser extent 6 are enriched for both these genes (FIG. 25 A), and may be functional and / or physical equivalents to CODEX fibroblast cluster 4 defined by high levels of ROBO2 and EID1 (FIG. 25B). It is possible that the bimodal expression of RoboZ and Eidl in scRNA-seq-defined fibroblast cluster 6 (FIG. 25A) could represent changes in proportion before and after EP300i treatment, indicative of fibroblasts in a transition state sensitive to chromatin remodelling. In comparison, scRNA-seq-defined fibroblast clusters 5 and 7 expanded in EP300i- treated dorsal wounds and facial wounds relative to PBS-treated dorsal wounds (FIG. 17E, 17F), providing additional evidence that RoboZ and Eidl underlie the fibroblasts’ regenerative capacity. Collectively, these data indicate that ROBO2 and EID1 are spatially enriched at a protein and an RNA level in wounds that heal with reduced fibrosis.

[0278] Chromatin profiling downstream of ROBO2-EID1-EP300 axis

[0279] In contrast to the notable shift in scRNA-seq fibroblast profiles between PBS- treated and EP300i-treated dorsal wounds (FIG. 17F, bottom middle vs. bottom right), the differences in chromatin accessibility in fibroblasts of PBS-treated and EP300i-treated dorsal wounds were less pronounced (FIG. 22C, bottom middle vs. bottom right / FIG. 38C,Il lbottom middle vs. bottom right). Because open chromatin is marked by several different histone modifications (H3K27ac, H3K4mel, H3K4me3, etc).49specific chromatin changes downstream of EP300 may not have been detectable via a global chromatin profiling assay such as ATAC-seq. It was hypothesized that a more direct interrogation of changes in EP300- dependent chromatin marks may be needed. Given the importance of the histone acetyltransferase EP300 in mediating wound fibrosis downstream of SLIT2-ROBO2, focus was place on assaying levels of histone 3 lysine 27 acetylation (EI3K27ac). which is a marker of EP300 activity and active enhancers.32,50

[0280] To quantify H3K27ac levels downstream of EP300, we performed CUT&Tag chromatin profiling of H3K27ac in primary mouse fibroblast cultures was performed and A. coll DNA was used for signal normalization.51H3K27ac levels were higher in dorsal fibroblasts compared to facial fibroblasts, both globally across the mouse genome (FIG. 26 A, - I- CBP112) and locally around 5 ’-end regions of genes active in fibroblasts (FIG. 26B, - 1- CBP112). Differential analy sis of gene-specific H3K27ac levels showed that dorsal fibroblasts maintained higher levels of H3K27ac across more fibrosis-related genes (Collagens. Fibulinl, Lysyl oxidase like 1. Platelet-derived growth factor receptors) than facial fibroblasts (FIG. 26C). Furthermore, H3K27ac associated with Robo2 and Eidl was decreased in dorsal compared to facial fibroblasts (FIG. 26C, left), consistent with their decreased expression levels in dorsal relative to facial fibroblasts (FIGS. 5E, 10A). Smallmolecule I-CBP112-mediated inhibition of EP300 activity significantly reduced H3K27ac globally and over fibrosis-associated genes in both cultured dorsal and facial fibroblasts (FIGS. 26A, 26B, + I-CBP112).

[0281] I-CBP112 is a potent and specific inhibitor of not only EP300 but also CREB- binding protein (CBP)38, another acetyltransferase. To more specifically assay H3K27ac downstream of the ROBO2-EID1-EP300 signaling axis, a similar chromatin profiling strategy was used, CUT&RUN,52 on CRISPR-mediated Eidl-knockout and control mouse fibroblasts cultured from dorsal or facial unwounded skin. E. coli spike- in DNA was used for signal normalization among samples. Depriving cells of Eidl increased H3K27ac globally across the genome (FIG. 26D) and around 5’-end regions of fibrosis-related genes (FIG. 26E) in both dorsal and facial fibroblasts, compared to the fibroblasts with preserved Eid\ . Specifically, Eidl knockout increased H3K27ac in facial fibroblasts to levels similar to that observed in control dorsal fibroblasts, both globally across the genome and near fibrosis-related genes (FIGS. 26D, 26E). Differential analysis of gene-specific H3K27ac showed significantly higher levels of H3K27ac associated with collagen genes in Eidl-knockout facial fibroblasts compared to control facial fibroblasts (FIG. 26F). Overall, the chromatin profiling data revealed that the reduced fibrogenic potential of facial fibroblasts is at least in part maintained by a more transcriptionally silent epigenetic state with decreased histone acetylation downstream of the ROBO2-EID1-EP300 axis, resulting in lower expression of ECM genes (FIG. 26G).

[0282] SLIT2, ROBO2, and EID1 are differentially expressed in human skin

[0283] To assess the translational potential of our findings to humans, we analyzed human skin samples derived from the face and the back. At baseline, unwounded human back skin contained thicker and denser collagen bundles and less prominent adnexal structures compared to unwounded human facial skin (FIG. 27A, left). The histological differences in the dermis were further accentuated within scars, which demonstrated much thicker collagen bundles more uniformly organized in an east-west orientation in the back than in the face (FIG. 27A, right). Consistent with our findings in mice (FIG. 15B), human un ounded skin and scars from the face exhibited significantly higher levels of ROBO2 and EID1 by IHC than unwounded skin and scars from the back (FIG. S27B). Furthermore, to investigate whether ROBO2 signaling may be more active in human tissues with more regenerative and less scar-forming potential, we compared existing single-cell fibroblast transcriptomes of unwounded second trimester human fetal skin60with those from unwounded human adult skin61,62(FIGS. S27C-27E). We found that overall, human fetal skin fibroblasts expressed statistically higher levels of both SLIT2 and ROBO2 than human adult skin fibroblasts (FIGS. S27F, 27G). The enrichment of SLIT2 and ROBO2 was especially pronounced in fibroblast clusters (4 and 5) with the highest proportions of fetal fibroblasts (FIGS. S27C, 27E, 27F). Given the ability of early fetal skin to heal scarlessly,63these results buttress our findings in mice showing that elevated expression of Slit2 and Robo2 is important for w ound healing with reduced scarring. Although we were not able to detect Eid J and Ep3()0 expression in these human datasets, this scRNA-seq data, along with our IHC analyses of human facial and dorsal skin, reveal that anatomically variant expression of components of the SLIT2-ROBO2-EID1 axis is conserved from mice to humans, and may also underlie the disparate scarring potential in human skin.DISCUSSION

[0284] It has long been known that not all fibroblasts are created equal and their developmental origins matter to their context-specific function. Embryologically determined topographic differentiation of human fibroblasts leads to distinct geneexpression profiles according to anatomical location.12,13Many of these topographically expressed genes are associated with site-specific anomalies seen in genetic diseases such as Ehler-Danlos and hand-foot-genital syndromes.12Using the novel four-site murine dermal injury model herein, it was shown that the ontology of fibroblasts also matters for adult wound healing: Neural-crest-derived facial fibroblasts differ from fibroblasts of other body sites by their intrinsic capacity to promote healing with less fibrosis. Further, we demonstrated that dorsal fibroblasts can be modulated towards a neural-crest-like state to produce a less fibrotic phenotype, highlighting their plasticity despite their embryologically determined fibrogenic potential.

[0285] Underlying the less fibrotic potential of neural-crest-derived facial fibroblasts is SLIT2-ROBO2 signaling, which maintains a repressed chromatin landscape and a less differentiated state via EID 1 -mediated inhibition of EP300. SLIT-ROBO signaling was originally identified as a guidance cue in axon branching and neuronal migration.53More recent reports have shown SLIT2 to be associated with organ regeneration including of the intestine and the lung.54,55In the skin, fibroblasts secrete SLIT2 to inhibit human fibrocyte differentiation.56In support of these data, it was found, as detailed herein, that upregulation of SLIT2-ROBO2 signaling in neural-crest-derived facial fibroblasts is both necessary and sufficient for the reduced fibrosis observed in facial cutaneous wounds.

[0286] scRNA-seq analysis identified Eidl as a downstream target of SLIT2-ROBO2 signaling in facial wound repair. EID1 is known to play a crucial role in myocyte and neural stem cell differentiation.35In the skin, fibroblasts with less scar forming behavior also express Eidl .15The in vitro and in vitro modulation of Eidl expression in fibroblasts herein showed that it is necessary and sufficient to reduce fibrosis in wounds. EID1 regulates differentiation and transcriptional activation through inhibition of EP300, a histone acetyltransferase that modulates expression of genes via chromatin remodelling.57The data herein show that expression of Eidl by Robo2+ fibroblasts was associated with a reduced fibrotic state. Consistent with this, hte chromatin profiling herein found higher I43K27ac levels in dorsal than in facial fibroblasts, both globally across the genome and more specifically associated with fibrosis-related genes. It is conceivable that unlike facial fibroblasts, dorsal fibroblasts are epigenetically primed at baseline to express higher levels of fibrotic ECM components and thus engender more scar formation. These epigenetic differences were maintained in culture after multiple cell passages, demonstrating the epigenetic stability of fibroblasts of disparate embryonic origins. Additionally, our spatial transcriptome and protein profiling revealed changes not only in fibroblast subpopulationsbut also in specific cell-cell interactions within profibrotic versus pro-regenerative wound milieu. Given the importance of fibroblast crosstalk with each other and with other cell types (e.g., immune cells), it is not surprising that alteration or manipulation of a relatively small percentage of fibroblasts in vivo (FIG. 10B) can effect significant changes in wound healing outcomes. How much of this occurs via direct cell-cell interactions versus paracrine signaling, and which factors are involved, are subjects for future investigation.

[0287] To pave the way towards clinical translation, a small-molecule EP300 inhibitor was used to mimic the role of EID1 in reducing fibrosis following skin injury. Pharmacological inhibition of chromatin co-regulatory factors has shown to be a selective and effective strategy to modulate oncogenic signaling, for example in multiple myeloma.58In the context of injury response, studies have also shown bromodomain proteins to be important in promoting fibrosis in the liver and lung.59,60In this study, bromodomain inhibition of EP300 was found to be an effective anti-fibrotic therapeutic target allowing dorsal scarring wounds to heal like facial wounds with less fibrosis. In line with this, our chromatin profiling in EP300i -treated cells demonstrated markedly reduced H3K27ac levels indicative of global chromatin repression, including around ECM genes. Furthermore, such changes in the epigenetic landscape were specific to EID1 -mediated EP300 inhibition and unlikely due to EP300i’s off-target effects, as similar trends in H3K27ac were observed by CRISPR-mediated Eidl knockout.

[0288] In summary, the reduced fibrotic neural-crest-like identity of adult facial fibroblasts is underpinned by the ROBO2-EID1 -EP300 axis, which can be pharmacologically activated to reduce scar formation during wound healing. This revealed that the embryologically predetermined fibrogenic capacity of fibroblasts can be modulated as a novel anti-scarring therapeutic approach. This study expands our understanding of cranial neural crest cells and their derivatives beyond their roles in facial morphogenesis, demonstrating that lessons learned from development can be exploited in adult animals to improve scarring in wound healing.

Claims

CLAIMS1. A method of stimulating wound healing in a subject in need thereof comprising administering an effective amount of an EP300 modulator.

2. The method of claim 1, wherein the EP300 modulator is chosen from:(i) SLIT2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) R0B02 or a functional variant thereof; and(iv) an EP300 inhibitor, an analog thereof, a derivative thereof or of an analog thereof; or a pharmaceutically acceptable salt of the foregoing.

3. The method of claim 2. wherein the EP300 modulator is an EP300 inhibitor comprising one or a combination of EP3001. Y08197, CCS1477, CPI-1612, DS17701585. DS-9300, or NEO3734, or a pharmaceutically acceptable salt thereof.

4. The method of any of claims 1 through 3, wherein the modulator of acetyltransferase is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof.

5. The method of any of claims 1 through 4, wherein the subject is a mammal.

6. The method of any of claims 1 through 5, wherein the subject is a human.

4. A method of treating or preventing fibrosis in a subject in need thereof comprising administering an effective amount of an EP300 modulator to the subject.

5. The method of claim 4, wherein the EP300 modulator is chosen from:(i) SLIT2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) R0B02 or a functional variant thereof; and(iv) an EP300 inhibitor, an analog thereof, a derivative thereof or of an analog thereof;or a pharmaceutically acceptable salt of the foregoing.

6. The method of claim 5, wherein the EP300 modulator is an EP300 inhibitor comprises at least one of EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS-9300, or NEO3734, or a pharmaceutically acceptable salt thereof.

7. The method of any of claims 4 through 6, wherein the modulator of acetyltransferase is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof.

8. The method of any of claims 4 through 7, wherein the subject is a mammal.

9. The method of any of claims 4 through 8, wherein the subject is a human.

10. A method of reducing severity of scarring or preventing scarring in a subject in need thereof comprising administering an effective amount of an EP300 modulator to the subject.

11. The method of claim 10, wherein the EP300 modulator is chosen from:(i) SL1T2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) ROBO2 or a functional variant thereof; and(iv) an EP300 inhibitor, an analog thereof, a derivative thereof or of an analog thereof; or a pharmaceutically acceptable salt of the foregoing.

12. The method of claim 11, wherein the EP300 modulator is an EP300 inhibitor comprising one or a combination of: EP300i, Y08197. CCS1477, CPI-1612, DS17701585, DS-9300, or NEO3734, and a pharmaceutically acceptable salt thereof.

13. A method of inhibiting acetyltransferase in a subject in need thereof comprising administering to the subject an EP300 modulator.

14. The method of claim 13, wherein the EP300 modulator is chosen from:(i) SLIT2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) R0B02 or a functional variant thereof; and(iv) an EP300 inhibitor, a derivative thereof or an analog thereof; or a pharmaceutically acceptable salt of the foregoing.

15. The method of claim 11, wherein the EP300 inhibitor comprises at least one of EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS-9300, or NEO3734, or a pharmaceutically acceptable salt thereof.

16. The method of any of claims 13 through 15, wherein the EP300 modulator is administered by oral adminstration. parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof.

17. The method of any of claims 13 through 16, wherein the subject is a mammal.

18. The method of any of claims 13 through 17, wherein the subject is a human.

19. A method of differentiating a fibroblast comprising contacting the fibroblast with an effective amount of an EP300 modulator.

20. The method of claim 19, wherein the EP300 modulator is chosen from:(i) SLIT2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) ROBO2 or a functional variant thereof; and(iv) an EP300 inhibitor, an analog thereof, a derivative thereof or of an analog thereof; or a pharmaceutically acceptable salt of the foregoing.

21. The method of claim 20, wherein the EP300 modulator is an EP300 inhibitor comprising one or a combination of: EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS- 9300, or NEO3734, and a pharmaceutically acceptable salt thereof.

22. The method of any of claims 19 through 21, wherein the method is performed in vitro.

23. The method of any of claims 19 through 21. wherein the method is performed in vivo within a mammal and further comprising administering to the mammal an effective amount of the EP300 modulator.

24. A method of reducing a population of profibrotic fibroblasts in a subj ect in need thereof comprising administering to the subject an EP300 modulator.

25. The method of claim 16, wherein the EP300 modulator is chosen from:(i) SLIT2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) R0B02 or a functional variant thereof; and(iv) an EP300 inhibitor, an analog thereof, a derivative thereof or of an analog thereof; or a pharmaceutically acceptable salt of the foregoing.

26. The method of claim 25. wherein the EP200 modulator is an EP300 inhibitor chosen from one or a combination of: EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS-9300, or NEO3734, and a pharmaceutically acceptable salt thereof.

27. The method of any of claims 24 through 26. wherein the EP300 modulator is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof.

28. The method of any of claims 24 through 27, wherein the subject is a mammal.

29. The method of any of claims 24 through 28, wherein the subject is a human.

30. A method of enhancing SOXIO and / or SOX9 expression in a wound of a subject in need thereof comprising administering to the wound of the subject an EP300 modulator.

31. The method of claim 30, wherein the EP300 modulator is chosen from:(i) SLIT2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) ROBO2 or a functional variant thereof; and(iv) an EP300 inhibitor, a derivative thereof or an analog thereof; or a pharmaceutically acceptable salt of the foregoing.

32. The method of claim 31, wherein the EP200 modulator is an EP300 inhibitor comprising one or a combination of: EP300i. Y08197, CCS1477, CPI-1612, DS17701585, DS- 9300, or NEO3734, and pharmaceutically acceptable salts thereof.

33. The method of any of claims 30 through 32, wherein the EP300 modulator is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof.

34. The method of any of claims 30 through 33, wherein the subject is a mammal.

35. The method of any of claims 30 through 34, wherein the subject is a human.

36. A pharmaceutical composition comprising: (i) an effective amount of an EP300 modulator or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

37. The pharmaceutical composition of claim 36, wherein the effective amount is from about 10 grams to about 1000 grams of the EP300 modulator.

38. The pharmaceutical composition of claim 36, wherein the EP300 modulator is chosen from one or a combination of:(i) SLIT2 or a functional variant thereof;(ii) EID1 or a functional variant thereof;(iii) ROBO2 or a functional variant thereof; and(iv) an EP300 inhibitor, a derivative thereof, an analog thereof; or a pharmaceutically acceptable salt of the foregoing.

39. The pharmaceutical composition of claim 38. wherein the EP300 modulator is an EP300 inhibitor chosen from: EP300i, Y08197, CCS1477, CPI-1612, DS17701585, DS- 9300, NEO3734, and pharmaceutically acceptable salts thereof.

40. The pharmaceutical composition of claim 36, wherein the EP300 modulator is an agent that increases expression of SLIT2, ROBO2 and / or EID1.

41. An isolated fibroblast comprising, or cell line comprising a fibroblast that comprises, Robo2 or a functional variant thereof.

42. The fibroblast or cell line of claim 41, further comprising one or a combination of: Cdh4+, Ncaml+, and Gpc3 or a functional fragment thereof.

43. The fibroblast or cell line of claim 27. wherein the fibroblast comprises SOX9 and / or SOXIO.

44. The fibroblast or cell line of any of claims 41 through 43, wherein fibroblast or cell line is frozen at about -20 degrees Celsius or about -212 degrees Celsius.

45. A pharmaceutical composition comprising an effective amount of fibroblasts of any of claims 41 through 44; and a pharmaceutically acceptable carrier.

46. A method of preventing or treating fibrosis in a subject in need thereof comprising administering to the subject an effective amount of the pharmaceutical composition of claim 45 or the fibroblasts of any of claims 41 through 44.

47. The method of claim 46, wherein the pharmaceutical composition of claim 45 or the fibroblasts of any of claims 41 through 44 is administered by oral adminstration, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof.

48. The method of any of claims 46 through 47, wherein the subject is a mammal.

49. The method of any of claims 46 through 48, wherein the subject is a human.

50. A method of promoting wound healing in a subject in need thereof comprising administering to the subject an effective amount of the pharmaceutical composition of claim 45 or the fibroblasts of any of claims 41 through 44.

51. The method of claim 50, wherein the pharmaceutical composition of claim 45 or the fibroblasts of any of claims 41 through 44 is administered by oral adminstrati on, parenteral administration, sublingual administration, transdermal administration, rectal administration, transmucosal administration, topical administration, inhalation, buccal administration, intrapleural administration, intravenous administration, intraarterial administration, intraperitoneal administration, subcutaneous administration, intramuscular administration, intranasal administration, intrathecal administration, and intraarticular administration, or combinations thereof.

52. The method of any of claims 50 through 51, wherein the subject is a mammal.

53. The method of any of claims 50 through 52, wherein the subject is a human.

54. The method of claim 50 wherein the pharmaceutical composition is administered directly into the wound of the subject.

55. The method of any of claims 50 through 54, wherein the method comprises administering from about 10,000 to about 10,000,000 cells of any of claims 41 through 44.

Images