Treatment of ulcerative colitis and symptoms thereof

Abstract

Methods of treating UC and IBD are disclosed herein. Particularly, it has been found that by blocking the redox activity of APE1 / Ref-1, the elevated expression of HIF-1α in HSPCs is inhibited and the dynamics of HSPCs both in normal and Dnmt3a mutant mice under inflammatory milieu of UC is reversed. It has further been found that blocking the redox activity of APE1 / Ref-1 is useful for treating IBD. The use of an APE1 / Ref-1 inhibitor in a medicament for treating UC and IBD are disclosed herein.

Description

[0001] 29715.00029 (2024-027-02) PCT TREATMENT OF ULCERATIVE COLITIS AND SYMPTOMS THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority and the benefit under 35 U.S.C. § 119(e) to U.S. Provisional Application No.63 / 598,589, filed on November 14, 2023, which is incorporated by reference in its entirety. BACKGROUND OF THE DISCLOSURE The present disclosure relates generally to the methods of treating ulcerative colitis (UC). Particularly, it has been found herein that by blocking the APE1 pathway, and particularly, the redox-activity of APE1 / Ref-1, through the administration of an apurinic / apyrimidinic endonuclease 1 redox factor 1 (APE1 / Ref- 1) inhibitor (e.g., APX3330), the expression of HIF-1α in hematopoietic stem and progenitor cells (HSPCs; also referred to as hematopoietic stem cells (HSCs)) isinhibited and the dynamics of HSPCs is reversed. Particularly, it has now been foundthat chronic UC contributes to the differentiation of HSPCs toward myelopoiesis in an APE1 / Ref-1 / HIF-1α / IL-1r1 pathway-dependent manner. Ulcerative colitis (UC) is a chronic recurring inflammatory disease of the gastrointestinal (GI) tract. While UC can occur across all ages, studies suggest that approximately 20% of patients with UC present before the age of 20 years, and the incidence rate continues to rise in the U.S. The etiopathogenesis of UC development is unclear; however, accumulating data suggests a role for the pathological immune response against microbial and environmental antigens in genetically predisposed individuals. It appears that both innate and adaptive immune systems in response to microbiota are involved in maintaining intestinal homeostasis. Studies have advocated that aberrant immune cells and their cytokines are thought to increase intestinal permeability by disrupting intestinal epithelial integrity. All the blood cells and immune cells are consistently replenished lifelong by a rare population of hematopoietic stem cells (HSCs) in a hematopoietic hierarchical manner in the bone marrow (BM). HSCs must maintain self-renewal and 29715.00029 (2024-027-02) PCT differentiation potential over time to preserve the hematopoiesis of the hematopoietic system. In the steady-state, HSCs are relatively quiescent; however, inflammatory conditions generated due to infection or stress, can activate HSCs to meet the demand of total blood cell pool called “emergency hematopoiesis”. The fate of HSCs renewal and differentiation is tightly controlled by both cell-intrinsic and cell-extrinsic factors. Under stress-induced tissue homeostasis, infection, and inflammatory conditions, HSCs can give rise to various lineage-restricted hematopoietic stem and progenitors (HSPCs) through the pool of intermediate hematopoietic cells called multipotent progenitors (MPPs). HSPCs are further stepwise differentiated into various subtypes of blood cells, including granulocytes, monocytes, and lymphocytes, which participate in the host immune response. The interaction between evolutionary conserved pathogen-associated molecular patterns (PAMPs) of bacteria and viruses and pathogen recognition receptors (PRRs), e.g., Toll-like receptors (TLRs) and the nucleotide-binding oligomerization domain-like receptors (NLRs) expressed by HSCs may lead to “emergency myelopoiesis”. Infection-induced chronic inflammation with a variety of pathogenic organisms, such as bacteria, viruses, and parasites, can result in profound alternations in hematopoiesis in the BM, including expansion of early progenitor compartment (Lineage-c-Kit+ Sca1+; also called LSK cells), changes in HSC differentiation and long-term reconstitution properties and migration patterns. Recently, gut microbiota have emerged to play a critical role in the manifestation and maintenance of many diseases, including UC, rheumatic arthritis, metabolic syndrome, neurodegeneration, and malignancy. However, several studies suggest that intestinal microbiota serve as a key regulator of hematopoiesis as germ- free and antibiotic-treated mice presented increased pathogen burden and a decrease in the HSPC pool, suggesting some of the responses associated with diverse infections are context dependent which, likely reflect different host-pathogen interactions. Genetic mutations in several genes have been associated with clonal hematopoiesis. DNA methyltransferase 3A (DNMT3A) is the most commonly mutated gene among epigenetic regulator genes such as DNMT3A, TET2, and 29715.00029 (2024-027-02) PCT ASXL1 in clonal hematopoiesis, and is associated with the risk of developing hematological malignancies like myelodysplastic syndrome (MDS). Dysregulated immune and inflammatory signaling pathways have been implicated in hematopoietic abnormalities related to aging, cardiovascular disease leading to MDS development. Although clonal hematopoiesis has been most intensively studied, only recently, an epidemiological sequencing study showed that the inflammatory environment of ulcerative colitis promoted positive selection of hematopoietic clones with DNMT3A mutation. Microbial infections drive pro- inflammatory cytokines, interferons (IFN-α / γ), tumor necrosis factor (TNF)-α, interleukin (IL)-1, IL-6, growth factors (G-CSF / M-CSF). These proinflammatory cytokines and bacterial endotoxins (e.g., LPS) released by leaky and inflamed gut enter into systemic circulation, causing inflammatory stress conditions in the bone marrow (BM). However, it remains largely unknown how pathophysiologic conditions of chronic ulcerative colitis contribute to hematopoiesis in normal and in Dnmt3a mutation settings and what underlying molecular mechanism(s) involved in the regulation of hematopoiesis, and whether and how hematopoietic defects could be corrected to its normalcy both in normal and mutation settings. Thus, at a more systems level, how signals that originate in the gut get translated into involving cells of the innate and adaptive immune system as well as those in the BM that give rise to these cells at the very primitive level are not known. Importantly, whether there is a way to reverse this systemic tissue based global changes that initially originate in the colon is not known. The present disclosure demonstrates that APE1 / Ref-1 / HIF-1α / IL-1r1 signaling pathway plays an important role in aberrant hematopoiesis and that APE- 1 / Ref-1 / HIF-1α / IL-1r1 signaling, and thus, could serve as a potential therapeutic target for chronic UC. Further, the blockade of the APE1 / Ref-1 / HIF-1α pathway mitigates the underlying pathological conditions of chronic UC and rescues aberrant hematopoiesis in normal and mutational settings of Dnmt3a. 29715.00029 (2024-027-02) PCT BRIEF DESCRIPTION The present disclosure relates generally to methods of treating UC in subjects in need thereof. Particularly, it has been found herein that by blocking the redox activity of APE1 / Ref-1, through the administration of APX3330 (and / or analogs thereof), the elevated expression of HIF-1α in HSPCs is inhibited and the dynamics of HSPCs both in normal and Dnmt3a mutant mice under inflammatory milieu of UC is reversed. The APE1 / Ref-1 inhibitor treatment also inhibited pro- inflammatory Ly6Chi monocytes. Based on the foregoing, in one aspect, the present disclosure is directed to a method of treating ulcerative colitis (UC)) in a subject in need thereof, the method comprising administering to the subject an effective amount of an apurinic / apyrimidinic endonuclease 1 redox factor 1 (APE1 / Ref-1) inhibitor, pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof. BRIEF DESCRIPTION OF THE DRAWINGS FIGS.1A-1GG show APE-1 / Ref-1 inhibitor, APX3330, rescues chronic UC mediated hematopoietic defects. FIGS.1A and 1B depict an experimental scheme illustrating the treatment protocol for the APX3330 drug and DSS (3%, w / v). Mice were gavaged with the drug APX3330 at 50 mg / Kg, mouse body weight, two times a day. The treatment regimen was started 7 days before the initiation of DSS treatment and continued throughout the cycles of DSS treatment. Veh – vehicle control, DSS+Veh, and APX3330+DSS. FIG. 1C depicts the total number of bone marrow (BM) cells and FIG.1D depicts the frequency of LSK cells (Lin-Sca-1+c- Kit+). FIG. 1E depicts the total bone marrow (BM) cells in DSS treated and untreated control mice. FIG. 1F shows representative flow cytometry profiles showing gating of bone-marrow (BM) Lineage (Lin)-, Sca-1(S)+, c-Kit (K)+(LSK cells, left panel) and characterization of LSK various hematopoietic stem cell subsets (right panel). FIG. 1G depicts the frequency of LSK, Lin-Sca-1+c-Kit+cells in the BM of DSS treated and non-DSS treated control mice. FIG. 1H depicts gating strategy for LSK’s various hematopoietic stem cell subsets. FIG.1I depicts the frequency of LT-HSCs (Lin-Sca- 29715.00029 (2024-027-02) PCT 1+c-Kit+CD150+CD48-). FIG.1J shows frequencies of LT-HSCs, CD150+CD48- among LSK cells in the BM of DSS treated and non-DSS treated control mice. FIG. 1K depicts the frequencies of MPPs or ST-HSCs (Lin-Sca-1+c-Kit+CD150-CD48-). FIG.1L depicts the frequencies of MPPs, CD150-CD48- among LSK cells in the BM of DSS treated and non-DSS treated control mice. FIG.1M depicts the frequencies of myeloid-biased hematopoietic progenitor (HPC2) cells (Lin-Sca-1+c- Kit+CD150+CD48+). FIG.1N depicts frequencies of HPC2, CD150+CD48+ among LSK cells in the BM of DSS treated and non-DSS treated control mice. FIG.1O shows spleen pictures of veh, DSS+veh, and APX3330+DSS treated mice. FIG. 1P depicts spleen (SP) weight of veh, DSS+veh, and APX3330+DSS treated mice. FIG. 1Q depicts total number of splenocytes in of veh, DSS+veh, and APX3330+DSS treated mice. FIG.1R depicts spleen images, FIG.1S depicts spleen (SP) weight in grams (gm), and FIG.1T depicts SP cellularity in DSS treated and control mice. FIG. 1U depicts total number of splenocytes in in DSS treated and control mice. FIGS. 1V, 1W, 1Y depict frequencies of LSKs, Lin-Sca-1+c-Kit+(FIG.1V) and LT-HSCs, CD150+CD48- (FIG.1W), HPC2, CD150+CD48+(FIG. 1Y) among LSK cells in the SP of DSS treated and non-DSS treated control mice. FIGS. 1X and 1Z-1CC depict frequencies of LSK cells (FIG.1X), of MPP cells (FIG.1Z), of granulocyte- macrophage progenitors, GMPs (Lin-Sca-1-c-Kit+CD16 / 32+CD34+) (FIG.1AA), of common-myeloid progenitors, CMPs (Lin-Sca-1-c-Kit+CD16 / 32loCD34+) (FIG.1BB), of megakaryocyte-erythrocyte progenitors, MEPs (Lin-Sca-1-c-Kit+CD16 / 32-CD34-) (FIG.1CC) in the BM of Veh, DSS+Veh, and APX3330+DSS treated mice. FIGS. 1DD-1FF depict the frequencies of LKs (FIG.1DD), of GMPs (FIG.1EE), and of MEPs (FIG.1FF) in the SP of Veh, DSS+Veh, and APX3330+DSS treated mice. FIGS.1GG & 1HH depict frequencies of Ly6Chiand Ly6Clomonocytes characterized by Ly6G; CD11b+; and Ly6Chiandloexpression in the BM and SP of Veh, DSS+Veh, and APX3330+DSS treated mice. Results are either representative data of two independent experiments or aggregate data of two independent experiments and each dot represents a single mouse. Data are shown as the mean ±SD. Statistical significance was determined using one-way ANOVA (FIGS. 1C-1FF) or two-way ANOVA (FIGS.1GG & 1HH) with Tukey’s multiple comparison test. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. 29715.00029 (2024-027-02) PCT FIG.2 depicts a schematic presentation of primary competitive bone- marrow transplantation (cBMT) assay. FIG.3 depicts a schematic presentation of primary competitive bone- marrow transplantation (cBMT) assay. FIGS.4A-4C show the absolute numbers of LSKs (FIG.4A), LT- HSCs (FIG.4B), and HPC2 (FIG.4C) among LSK cells in the BM of control and DSS treated mice). FIGS.4D-4F show the absolute numbers of LSKs (FIG.4D), LT- HSCs (FIG.4E), and HPC2 (FIG.4F) in the BM of Veh, DSS+Veh, and APX3330+DSS treated mice. FIGS.4G-4J depict the absolute numbers of LSKs (FIG.4G), LT- HSCs (FIG.4H), MPPs (FIG.4I), and HPC2 (FIG.4J) in the SP of control and DSS treated mice. FIGS.4K-4N depict the absolute numbers of LSKs (FIG.4K), LT- HSCs (FIG.4L), MPPs (FIG.4M), and HPC2 (FIG.4N) among LSK cells in the SP of Veh, DSS+Veh, and APX3330+DSS treated mice. FIGS.5A-5T show that the APE-1 / Ref-1 inhibitor, APX3330, rescues chronic UC mediated hematopoietic defects. Particularly, FIG.5A shows flow- cytometric analysis of bone-marrow (BM) immature Lin-, c-Kit+Sca-1- (LK cells, left panel) and various hematopoietic stem and progenitor cell (HSPC) subsets (right panel). FIGS.5B-5F depict the frequencies of immature LKs, Lin-c-Kit+Sca-1- (FIG. 5B), of granulocyte-macrophage progenitors GMPs, LKgatedCD16 / 32+CD34+(FIG. 5C), of common-myeloid progenitors CMPs, LKgatedCD16 / 32loCD34+(FIG. 5D), of megakaryocyte-erythroid progenitors MEPs, LKgatedCD16 / 32-CD34- (FIG.5E), and of common-lymphoid progenitors CLP, Lin-CD127+c-KitloSca-1lo(FIG. 5F) in the BM of DSS treated and non-DSS treated control mice. FIGS.5G-5J show the absolute numbers of immature LKs, Lin-c-Kit+Sca-1- (FIG.5G), GMPs, LKgatedCD16 / 32+CD34+(FIG.5H), CMPs, LKgatedCD16 / 32loCD34+(FIG.5I), and 29715.00029 (2024-027-02) PCT CLP, Lin-CD127+c-KitloSca-1lo(FIG. 5J) in the BM of DSS treated and non-DSS treated control mice. FIGS.5K-5O depict the frequencies of immature LKs, Lin-c- Kit+Sca-1- (FIG.5K), and GMPs, LKgatedCD16 / 32+CD34+(FIG. 5L), and the absolute numbers of immature LK, Lin-c-Kit+Sca-1- (FIG.5M), GMPs, LKgatedCD16 / 32+CD34+(FIG. 5N), and CMPs, LKgatedCD16 / 32loCD34+(FIG.5O) in the SP of DSS treated and non-DSS treated control mice. FIGS.5P-5T show the absolute numbers of immature LKs, Lin-c-Kit+Sca-1- (FIG.5P), GMPs, LKgatedCD16 / 32+CD34+(FIG. 5Q), and CLP, Lin-CD127+c-KitloSca-1lo(FIG.5R) in the BM and LKs, Lin-c-Kit+Sca-1- (FIG.5S), GMPs, LKgatedCD16 / 32+CD34+(FIG. 5T), in the SP of Veh, DSS+Veh, and APX3330+DSS treated mice. Results are pooled data of two independent experiments, and each dot represents a single mouse (B-O n=10, and P-T n=7-8 mice per group). Data are presented as the mean ±SD. Statistical significance was obtained using an unpaired, two-tailed Student’s t-test (B- O) or one-way ANOVA with Tukey’s multiple comparison test (P-T) for the analysis of differences between the experimental groups. **P<0.01, ***P<0.001 and ****P<0.0001; n.s., not significant. FIGS.6A-6Q show the APE-1 / Ref-1 inhibitor, APX3330, rescues chronic UC mediated hematopoietic defects. FIG.6A depicts flow-cytometric analysis of apoptosis of BM LSK cell by staining with 7-AAD and annexin-V. FIG. 6B depicts the frequency of BM LSK apoptotic (7-AAD+Annexin-V+) cells between DSS treated and non-DSS treated control mice. FIG. 6C is a representative flow-plot showing analysis of cell-cycle of BM LSK cells using DAPI staining. FIG.6D shows quantification of BM LSK cells in G0 / G1, S and G2 / M phase. FIG.6E depicts the changes in body weight and FIG.6F depicts the disease activity index (DAI) during 4 cycles of DSS treatment. FIG.6G are representative graphs for colon tissue, and FIG. 6H shows the quantification of colon length expressed in centimeters (cm) of DSS treated and untreated control mice. FIG.6I depict mice showing diarrhea and rectal bleeding with 3% DSS (see red arrow). FIGS.6J-6Q depict the frequencies of CD11b+Gr1+neutrophils (FIG. 6J), CD11b+F4 / 80+macrophages (FIG. 6K), CD11c+myeloid cells (FIG.6L), c-DC (CD11c+CD11b+B220-) and p-DC (CD11c+B220+CD11b-) (FIG.6M), B220+B cells (FIG.6N) and CD3+T cells (FIG. 29715.00029 (2024-027-02) PCT 6O), CD4+and CD8+T cells (FIG.6P), T reg cells (FIG. 6Q), among CD45+ leukocytes in the colon of DSS treated and untreated control mice. Results are either representative data of a single experiment (FIGS. 6B & 6D, n=5 mice per group), two independent experiments (FIGS. 6E-6F, n=5 mice per group) or cumulative data of two independent experiments (FIG.6H & FIGS. 6J-6Q, n=5-10 mice per group), and each dot represents a single mouse. Data are presented as the mean ±SD. Statistical significance was determined using either an unpaired, Student’s two-tailed t-test (FIGS.6B, 6D, 6H, 6J-6L, 6N-6O and 6Q) two-way ANOVA with Sidak’s multiple comparison test (E-F, & M & P) for the analysis of differences between the experimental groups. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.7A-7O show that the APE-1 / Ref-1 inhibitor, APX3330, rescues DSS-induced chronic UC associated phenotypes via inhibition of G-CSF, chemokine KC and hypoxia-inducible transcription factor-1α (HIF-1α). FIGS.7A is a representative graph presenting changes in body weight, FIG.7B depicts disease activity index (DAI) during 4 cycles of DSS treatment, FIG. 7C are images of colon tissue, and FIG.7D depict the quantification of colon length expressed in centimeters (cm) of Veh, DSS+Veh, and APX3330+DSS treated mice. FIGS.7E & 7F depict the frequency CD11b+Gr1+neutrophils (FIG.7E), and CD3+ T cells (FIG. 7F) in the colon of Veh, DSS+Veh, and APX3330+DSS treated mice. FIGS.7G-7L depict the frequencies of neutrophils (CD11b+Gr-1+) (FIG. 7G), B220+B cells (FIG.7H), and CD3+T cells (FIG.7I) in peripheral blood (PB) of Veh, DSS+Veh, and APX3330+DSS treated mice. Frequencies of neutrophils (CD11b+Gr-1+) (FIG.7J), B220+B cells (FIG. 7K), and CD3+T cells (FIG.7L) in the bone marrow (BM) of Veh, DSS+Veh, and APX3330+DSS treated mice. FIGS.7M-7O depict the frequencies of neutrophils (CD11b+Gr-1+) (FIG. 7M), B220+B cells (FIG.7N), and CD3+T cells (FIG.7O) in the spleen (SP) of Veh, DSS+Veh, and APX3330+DSS treated mice. Results are either representative data of two independent experiments (FIGS.7A-7F; n= 4 mice per group) or are cumulative data of two independent experiments (FIGS.7G-7O; n= 7-8 mice per group) and each dot represents a single mouse. Data are shown as the mean ±SD. Statistical significance was determined 29715.00029 (2024-027-02) PCT using two-way ANOVA (A-B, & D) or one-way ANOVA (G-O) with Tukey’s multiple comparison test. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.8A-8H show that APE-1 / Ref-1 inhibitor, APX3330, rescues DSS-induced chronic UC associated phenotypes via inhibition of G-CSF, chemokine KC and hypoxia-inducible transcription factor-1α (HIF-1α). FIG. 8A represents cytokine G-CSF concentration expressed in ng / ML, and chemokine KC expressed in pg / mL in Veh, DSS+Veh, and APX3330+DSS treated mice serum. FIG.8B is a flow- cytometric profile, showing gating strategy for pNF-ĸBp65+LK cells using intracellular staining. FIG.8C depicts the frequency of pNF-ĸBp65+LK (Lin-Sca-1-c- Kit+) cells from the BM of Veh, DSS+Veh, and APX3330+DSS treated mice. FIG. 8D depicts a flow-cytometric profile showing gating strategy for HIF-1α+LK cells using intracellular staining. FIGS.8E & 8F depict the frequencies of HIF-1α+LK (Lin-Sca-1-c-Kit+) cells from the BM (FIG.8E), and SP (FIG. 8F); and FIGS. 8G & 8H show the absolute numbers of HIF-1α+LK (Lin-Sca-1-c-Kit+) cells from the BM (FIG.8G), and SP (FIG. 8H). For FMO staining, an equal concentration of fluorophore matched isotype control antibody was used. Results are from a single experiment with mice (n=4) in each experimental group. Each dot represents a single mouse. Data are shown as the mean ±SD. Statistical analysis was performed using a one-way ANOVA with Tukey’s multiple comparison test. *P<0.05, and **P<0.01; n.s., not significant. FIGS.9A-9E depict APE-1 / Ref-1 inhibitor, APX3330, rescues DSS- induced chronic UC associated phenotypes via inhibition of G-CSF, chemokine KC and hypoxia-inducible transcription factor-1α (HIF-1α). FIG.9A represents various cytokine and chemokine analyses in sera of DSS untreated control and DSS treated mice. FIG. 9B is a schematic presentation of primary competitive bone-marrow transplantation (cBMT) assay. FIG.9C shows an analysis of PB at 4, 8, 12, and 16 weeks post-cBMT. FIGS.9D & 9E depict BM (FIG. 9D) and SP (FIG.9E) at 16 weeks post-cBMT showing engraftment of donor derived CD45.2 cells. Results are either pooled (FIG.9A, n=8-10) or representative data (FIGS. 9B-9E, n=5) of two independent experiments. Each dot represents a single mouse. Data are presented as 29715.00029 (2024-027-02) PCT the mean ±SD. Statistical analysis was performed using either an unpaired two-tailed Student’s t-test for the analysis of differences between the experimental groups (FIGS.9A, 9D & 9E) or two-way ANOVA with Sidak’s multiple comparison test (FIG.9C). *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001. FIGS.10A-10N show that the blockade of redox activity of APE- 1 / Ref-1 by its inhibitor APX3330 improves repopulating ability of chronic UC triggered defective HSC under transplant setting. Particularly, FIG.10A is a schematic presentation of a primary competitive bone-marrow transplantation (cBMT) assay. FIGS.10B & 10C depict the analysis of PB at 4, 8, 12,16, and 20 weeks post-cBMT (FIG.10B), and BM (FIG.10C) at 20-week post-cBMT showing engraftment of donor-derived CD45.2 cells. FIG.10D depicts total BM cells. FIGS. 10E-10H depict the frequencies of donor-derived LSK cells (FIG.10E), LT-HSCs (FIG.10F), MPPs (FIG. 10G), and HPC2 (FIG.10H) in the BM of F1 recipient mice transplanted with CD45.2+total BM cells of Veh, DSS+Veh, and APX3330+DSS mice. FIGS.10I-10K depict frequencies of donor-derived LKs (FIG.10I), GMPs (FIG.10J), and MEPs (FIG.10K) progenitors in the BM of F1 recipient mice transplanted with CD45.2+total BM cells of Veh, DSS+Veh, and APX3330+DSS mice. FIG.10L depicts the frequency of donor-derived neutrophils (CD11b+Gr1+) cells in the BM of F1 recipient mice transplanted with CD45.2+total BM cells of Veh, DSS+Veh, and APX3330+DSS mice. FIG.10M is representative graphs for colon tissue of F1 recipient mice transplanted with CD45.2+total BM cells of Veh, DSS+Veh, and APX3330+DSS mice, and FIG.10N shows the quantification of colon length expressed in centimeters (cm). Results are from a single experiment with mice (n=5) in each experimental group. Each dot represents a single mouse. Data are presented as the mean ±SD. Statistical significance was determined using either two- way ANOVA (FIG. 10B) or one-way ANOVA (FIGS.10C-10L & FIG.10N) with Tukey’s multiple comparison test for the analysis of differences between the experimental groups. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.11A-11P depict HIF-1α specific inhibitor, echinomycin, reduces accumulation of proinflammatory CD4+ T helper 1 (Th1) and Th17 cells in the colon 29715.00029 (2024-027-02) PCT by reducing IL-1β and IL-6 production. Mice were treated with echinomycin at 10 µg / Kg, mouse body weight, on alternate days through intraperitoneal injection. The treatment regimen started 7 days before the initiation of DSS treatment and continued throughout the cycles of DSS (3% w / v) treatment. FIG.11A showshanges in body weight, and FIG.11B depicts disease activity index (DAI) during 4 cycles of DSS treatment. FIG.11C are reprentative graphs for colon tissue of Veh, DSS+Veh, and DSS+Echin treated mice, and quantification of colon length expressed in centimeters (cm). FIG.11D are representative graphs for spleen (SP), FIG. 11E show SP weight expressed in gm, and FIG.11F show total number of splenocytes of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.11G depict frequency of CD4+T cells, FIG. 11H depict frequency of T reg cells, FIG.11I depict frequency of Th1 cells, FIG.11J depict frequency of Th17 cells, and FIG.11K depict ratio of Treg / CD4+T cells among CD45+leukocytes in the colon of Veh, DSS+Veh, and DSS+Echin treated mice. FIG. 11L depict the percentage of CD11b+myeloid cells, and FIG. 11M depict CD3+T cells expressing IL-1β, IL-6, and TNF-α cytokines among CD45+leukocytes in the colon of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.11N depict the percentage of GM-CSF+Th1 cells, FIG. 11O depict the percentage of G-CSF+Th17 cells, and FIG.11P depict GM-CSF+Th17 cells among CD45+leukocytes in the colon of Veh, DSS+Veh, and DSS+Echin treated mice. Results are from a single experiment with mice (n=5 to 8) in each experimental group. Each dot represents a single mouse. Data are shown as the mean ±SD. Statistical significance was determined using either two-way ANOVA (FIGS.11A & 11B & FIGS. 11M & 11N) or a one-way ANOVA (FIGS.11C-11L & FIGS.11O-11P) with Tukey’s multiple comparison test for the analysis of differences between the experimental groups. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.12A –12Z show that the HIF-1α specific inhibitor, echinomycin, restores chronic UC induced defective hematopoiesis. FIG.12A represents absolute BM cellularity of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.12B represents frequency of LSK cells in the BM of Veh, DSS+Veh, and DSS+Echin treated mice. FIGS.12C-12E depict the frequency of LT-HSCs (FIG.12C), frequency of MPPs (FIG.12D), frequency of HPC2 (FIG.12E) among LSK progenitors in the BM of 29715.00029 (2024-027-02) PCT Veh, DSS+Veh, and DSS+Echin treated mice. FIG. 12F depicts the frequency of LSK cells in the SP of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.12G depicts the frequency of LT-HSCs, and (FIG.12H) frequency of MPPs among LSK progenitors in the SP of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.12I depicts the frequency of LK cells in the BM of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.12J depicts the frequency of GMPs, FIG.12K depicts the frequency of CMPs, and FIG.12L depicts the frequency of MEPs among LK progenitors, and FIG. 12M depicts the frequency of CLP (Lin-CD127+c-KitloSca-1lo) in the BM of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.12N depicts the frequency of LK cells in SP of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.12O depicts the frequency of GMPs, FIG.12P depicts the frequency of CMPs, and FIG. 12Q depicts the frequency of MEPs among LK progenitors in the SP of Veh, DSS+Veh, and DSS+Echin treated mice. FIGS.12R-12Z depict the frequencies of CD11b+Gr1+neutrophils (FIG.12R), B220 B cells (FIG. 12S), and CD3+T cells (FIG.12T) in the peripheral blood (PB), CD11b+Gr1+neutrophils (FIG.12U), B220 B cells (FIG.12V), and CD3+T cells (FIG. 12W) in the BM, and CD11b+Gr1+neutrophils (FIG.12X), B220 B cells (FIG.12Y), and CD3+T cells (FIG.12Z) in the SP of Veh, DSS+Veh, and DSS+Echin treated mice. Results are from a single experiment with mice (n=5-6) in each experimental group. Each dot represents a single mouse. Data presented as the mean ±SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test for the analysis of differences between the experimental groups. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.13A-13Q depict HIF-1α specific inhibitor, echinomycin, reduces accumulation of proinflammatory CD4+T helper 1 (Th1) and Th17 cells in the colon by reducing IL-1β and IL-6 production. FIGS.13A-13D show the absolute numbers of LSKs (FIG.13A), LT-HSCs (FIG.13B), MPPs (FIG.13C), and HPC2 (FIG.13D) in the BM of Veh, DSS+Veh, and DSS+Echin treated mice. FIGS.13E- 13H show the absolute numbers of LSKs (FIG.13E), LT-HSCs (FIG.13F), MPPs (FIG.13G) and HPC2 (FIG.13H) in the SP of Veh, DSS+Veh, and DSS+Echin treated mice. FIGS.13I-13M show the absolute numbers of immature LKs (FIG. 13I), 29715.00029 (2024-027-02) PCT GMPs (FIG. 13J), CMPs (FIG.13K), MEPs (FIG.13L), and CLP (FIG.13M) in the BM of Veh, DSS+Veh, and DSS+Echin treated mice. FIGS.13N-13Q depict the absolute numbers of immature LKs (FIG.13N), GMPs (FIGS.13O), CMPs (FIG. 13P), and MEPs (FIG.13Q) in the SP of Veh, DSS+Veh, and DSS+Echin treated mice. Results are from a single experiment with mice (n=5-8) in each experimental group. Each dot represents a single mouse. Data represent mean ±SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.14A-14G depict that the HIF-1α specific inhibitor, echinomycin, restores chronic UC induced defective hematopoiesis by regulating Interleukin- 1receptor1 (IL-1r1) signaling in HSPCs. FIG.14A is a flow-cytometric profile showing gating strategy for HIF-1α+LK cells from the BM using intracellular staining (left panel) and the frequency of HIF-1α+LK (Lin-Sca-1-c-Kit+) cells (right panel), FIG.14B is a flow-cytometric profile showing gating strategy for HIF-1α+Lin-Sca-1-c-Kit+CD34+HSPCs from the BM using intracellular staining (left panel) and the frequency of HIF-1α+Lin-Sca-1-c-Kit+CD34+HSPCs (right panel). FIG.14C depicts the frequency of IL-1r1+LK (Lin-Sca-1-c-Kit+) cells, FIG.14D depicts the frequency of IL-1r1+Lin-Sca-1-c-Kit+CD34+HSPC cells from the BM of Veh, DSS+Veh, and DSS+Echin treated mice. FIG.14E is a flow-cytometric profile showing gating strategy for Ly6Chiand Ly6Clocells from the BM (left panel) and the frequencies of Ly6Chiand Ly6Clocells in the BM (right panel). FIGS.14F & 14G depict the frequencies of Ly6Chiand Ly6Clocells in the PB (FIG.14F), and spleen (FIG.14G) of Veh, DSS+Veh, and DSS+Echin treated mice. Results are from a single experiment with mice (n=5 to 6) in each experimental group. Each dot represents a single mouse. Data are presented as the mean ±SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test for the analysis of differences between the experimental groups (A-J). *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001; n.s., not significant. FIGS.15A-15P shows that chronic UC drives expansion of myelopoiesis, suppresses erythropoiesis, and profoundly impairs stem cell functions 29715.00029 (2024-027-02) PCT in pre-leukemic Dnmt3a mutant mice.8-10 weeks old Dnmt3afl / flMx1Cre+mice were injected with 3 doses of poly I:C (10µg / gm mouse body weight) to generate Dnmt3a mutant (D3a- / -) mice. Similarly, C57BL / 6 (B6) mice were also treated with poly I: C. After 2 weeks of poly I:C, D3a- / -and B6 mice were treated with and without 2% DSS (w / v) for 3 cycles of 5 days with DSS and 7 days of resting (drinking water) between each cycle. After the 3rd cycle of DSS, mice were sacrificed and analyzed for clonal hematopoiesis. FIG.15A depicts the total number of BM cells (pooled data from two independent experiments; n=7-9 mice per group). FIG.15B is representative flow cytometry profiles showing gating of BM LSK cells in B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIG.15C depicts the frequency of LSK cells in the BM of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS.15D-15F depict the frequencies of LT-HSCs (FIG.15D), MPPs (FIG.15E), and HPC2 cells (FIG.15F) within LSKs cells in the BM of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. (FIG.15G depicts the frequency of immature LKs in the BM of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS. 15H and 15I depict the frequencies of GMPs (FIG.15H), and MEPs (FIG.15I) within LKs in the BM of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS. 15J & 15K depict frequencies of Ly6Chiand Ly6Clomonocytes characterized by Ly6G; CD11b+; and Ly6Chiandloexpression in the BM and SP of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS.15L-15N show an analysis of PB chimerism of donor- derived CD45.2 cells at 4, 8, 12, 20 and 24 weeks post-cBMT. Analysis of chimerism of donor-derived CD45.2 cells at 24 weeks post-cBMT in the BM is shown in FIG. 15O, and in the SP in FIG. 15P of F1 recipient mice transplanted with CD45.2+ BM cells of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. Results are either aggregate data of two independent experiments (FIGS.15A, & FIGS.15J & 15K; n=6-10 mice per group) or the representative data of two independent experiments (FIGS.15C-15I; n=3-5 mice per group), or of a single experiment (FIGS. 15M-15P; n=5 mice per group), and each dot represents a single mouse. Data are shown as the mean ±SD. Statistical significance was determined using either one-way ANOVA (FIGS.15A, FIGS.15C-15I, & FIGS.15N-15P) or two-way ANOVA (FIGS. 15J & 15K, & FIG.15M) with Tukey’s multiple comparison test for the analysis of 29715.00029 (2024-027-02) PCT differences between the experimental groups. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.16A-16O depict chronic UC drives expansion of myelopoiesis, suppresses erythropoiesis, and profoundly impairs stem cell functions in pre-leukemic Dnmt3a mutant mice. FIGS.16A-16D depict the absolute numbers of LSK (FIG. 16A), LT-HSCs (FIG.16B), MPPs or ST-HSCs (FIG. 16C), and HPC2 (FIG.16D) in the BM of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS.16E depicts the absolute numbers of immature LKs, and FIG.16F depicts the absolute numbers of immature GMPs in the BM of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS.16G-16I depict the absolute numbers of immature LKs (FIG. 16G), GMPs (FIG.16H), and MEPs (FIG.16I) in the SP of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS.16J-16O show the frequency of CD11b+Gr-1+neutrophils in the BM (FIG. 16J) and SP (FIG.16K), frequency of B220+B cells in the BM (FIG.16L) and SP (FIG.16M), frequency of CD11b+Gr-1+neutrophils in the colon (FIG.16N), and frequency of CD3+T cells in the colon (FIG. 16O) of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. Results are representative data of two independent experiments with mice (n=3-) in each experimental group. Each dot represents a single mouse. Data represent mean ±SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.17A-17I depict chronic UC drives expansion of myelopoiesis, suppresses erythropoiesis, and profoundly impairs stem cell functions in pre-leukemic Dnmt3a mutant mice. Analysis of donor-derived HSCs functions in the BM of F1 recipient mice at 24 weeks post-TP. FIGS.17A-17D depict the frequencies of donor- derived LSKs (FIG. 17A), LT-HSCs (FIG.17B), MPPs (FIG.17C), and HPC2 (FIG. 17D) in the BM of F1 recipient mice transplanted with CD45.2+donor-derived cells of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIG.17E depicts the frequency of donor-derived MEPs progenitors in the BM of F1 recipient mice transplanted with CD45.2+donor-derived cells of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIG.17F depicts the frequency of donor-derived neutrophils 29715.00029 (2024-027-02) PCT (CD11b+Gr1+) in the BM of F1 recipient mice transplanted with CD45.2+donor- derived cells of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIG.17G depicts the frequency of donor-derived B220+B cells in the BM of F1 recipient mice transplanted with CD45.2+donor-derived cells of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. FIGS.17H & 17I depict the frequency of donor-derived myeloid-blast (CD11b+c-Kit+) cells in the BM and SP of F1 recipient mice transplanted with CD45.2+donor-derived cells of B6-control, B6+DSS, D3a- / --control, and D3a- / -+DSS mice. Results are from a single experiment with mice (n=4-5) in each experimental group. Each dot represents a single mouse. Data are presented as the mean ±SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test for the analysis of differences between the experimental groups. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIGS.18A-18P depict the APE1 / Ref-1 inhibitor, APX3330, restores chronic UC driven defective hematopoiesis and mitigates gut damage in B6 mice and D3a mutant mice in an APE1 / Ref-1 / HIF-1α axis dependent manner. FIG.18A is an experimental scheme illustrating the APX3330 and the cycles of DSS treatment. FIG. 18B are representative graphs for colon tissue of Veh, DSS+Veh, and DSS+APX3330 treated B6 and Dnmt3a- / - mice. FIG.18C depicts the quantification of colon length expressed in centimeters (cm). FIG. 18D shows the colon weight expressed in grams (gms). FIG.18E shows the colon weight / length ratio. FIGS.18F-18H depict the frequency of CD11b+Gr-1+neutrophils, FIGS.18I-18K depict the frequency of B220+B cells, and (FIGS.18L-18N) show the frequency of CD3+T cells in the PB, BM, and SP of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. FIGS. 18O & 18P depict the frequency of CD11b+c-Kit+myeloid blasts in the PB and the BM of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. Results are of a single experiment with mice (n=4-7) in each experimental group. Each dot represents a single mouse. Data represent mean ±SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. 29715.00029 (2024-027-02) PCT FIGS.19A-19P depict that the APE1 / Ref-1 inhibitor, APX3330, restores chronic UC driven defective hematopoiesis and mitigates gut damage in B6 mice and D3a mutant mice in an APE1 / Ref-1 / HIF-1α axis dependent manner. FIGS. 19A depicts the total number of BM cells of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. Frequencies of LSKs (FIG. 19B) in the BM of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. Frequency of MEPs (FIG. 19C) in the BM of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. FIG.19D shows the number of RBC, FIG.19E shows the concentration of HGB, FIG.19F shows the percentage of HCT, FIG.19G shows the percentage of RDW-CV, FIG.19H shows the number of neutrophils, FIG.19I shows the percentage of neutrophils, FIG.19J shows the number of lymphoid, FIG.19K shows the percentage of lymphoid cells in the PB of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. FIG.19L is representative graphs for spleen (SP), FIG.19M depicts SP weight in gms, FIG. 19N depicts SP cellularity. FIG.19O depicts the frequency of CD11b+c-Kit+myeloid blasts in the SP of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. FIG.19P depicts the frequency of HIF-1α expressing CD34+LK cells in the BM of Veh, DSS+Veh, and DSS+APX3330 treated B6 and D3a- / -mice. Results are of a single experiment with mice (n=4-7) in each experimental group. Each dot represents a single mouse. Data are presented as the mean ±SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test. *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001; n.s., not significant. FIG.20 is a schematic model showing possible mechanism of chronic UC induced hematopoietic defects in mice. DETAILED DESCRIPTION Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosure belongs. Although any methods and materials similar to or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are described below. 29715.00029 (2024-027-02) PCT The present disclosure relates generally to methods of treating ulcerative colitis (UC) in a subject in need thereof. Particularly, it has been found herein that by blocking the redox activity of APE1 / Ref-1, through the administration of APX3330 (and / or analogs thereof), elevated expression of HIF-1α in HSPCs is inhibited and the dynamics of HSPCs reversed. The APE1 / Ref-1 inhibitor treatment further inhibits pro-inflammatory Ly6Chimonocytes. Further, as APE1 / Ref-1 is a dual functioning protein that acts as an essential regulator of cellular responses to oxidative stress, which tress plays an important role in pathophysiological mechanisms involved in inflammation induced enteric neuronal loss and damage, blocking APE1 through administration of APX3330 further reduces oxidative stress, thereby further reducing inflammation and chronic pain. In suitable embodiments, the present disclosure includes administering to a subject in need thereof an effective amount of an APE1 inhibitor, pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof, the APE1 inhibitor capable of interacting with the APE1 protein such to cause unfolding of the APE1 protein in the amino terminal portion of APE1, inhibiting the ability of APE1 to interact with other proteins in the neurons or to perform its redox signaling function. More particularly, APE1 inhibitors used in the present disclosure have anti- inflammatory effects, blocking the ability of APE1 / Ref-1 to convert NF-κB and AP-1 from an oxidized state to reduced state, thereby altering their transcriptional activity. These inhibitors have been shown to suppress the production of pro-inflammatory cytokines and inflammatory mediators in murine macrophages. This results in the inability of NF-κB and AP-1 to bind to their target DNA sequence. Moreover, the inhibition allows direct down regulation of inflammatory cytokine secretion and ROS activation. Targeting the specific inhibition of APE1 / Ref-1 redox pathways and utilizing the DNA repair domain can lead to a UC therapy. More particularly, it has now been found that the APE1 / Ref-1 / HIF-1α / IL-1r1 signaling cascade plays a significant role in aberrant hematopoiesis that contributes to the pathophysiological conditions of chronic UC in a feed-forward loop manner. By targeting the APE1 / Ref- 1 / HIF-1α pathway, a better therapeutic target for ulcerative colitis can be obtained. 29715.00029 (2024-027-02) PCT Accordingly, in particular suitable embodiments, the APE1 inhibitor has the formula: R3YFormula (I) consisting of alkyl, alkoxy, hydroxyl, and hydrogen; R2 is an alkyl; R3 and R6 are independently selected from the group consisting of a substituted or unsubstituted alkoxy, a substituted or unsubstituted aryl and an oxo; R4 and R5 are independently selected from the group consisting of an alkoxy and aryl, or both R4 and R5 taken together form a substituted or unsubstituted napthoquinone; X is selected from the group consisting of CH=CR2and NCH, wherein R2is selected from the group consisting of C1-C10 alkyl and CF3CH2CH2; and Y is selected from the group consisting of N(Rz)R2 or NR^OR^, wherein each Rz is independently selected from the group consisting of C1-C6alkyl, heteroalkyl, cycloalkyl and cycloheteroalkyl, straight or branched chain or optionally substituted, or both Rz and R2 taken together with the attached nitrogen form an optionally substituted heterocycle; where each R^ is independently selected from the group consisting of hydrogen, alkyl, heteroalkyl, cyclohexyl, and cycloheteroalkyl, each of which is optionally substituted, or both R^ are taken together with the attached nitrogen and oxygen to form an optionally substituted heterocycle. Particularly suitable APE1 inhibitors include 3-[(5-(2,3-dimethoxy-6- methy11,4-benzoquinoy1)]-2-nony1-2-proprionic acid, (hereinafter "E3330" or "3330" or "APX3330"), and / or its analogs (e.g., [(2E)-2-[(3-methoxy-1,4-dioxo-1,4- dihydronaphthalen-2-yl)methylidene]-N,N-diethylpentanamide] (hereinafter 29715.00029 (2024-027-02) PCT "APX2009"), (2E)-2-[(3-methoxy-1,4-dioxo-1,4-dihydronapthalen -2- yl)methylidene]-N,N-dimethylpentanamide] (hereinafter "APX2007"), (2E)-2-[(3- methoxy-1,4-dioxo-1,4-dihydronapthalen -2-yl)methylidene]-N- methoxypentanamide] (hereinafter "APX2014"), (2E)-2-(3-methoxy-1,4-dioxo-1,4- dihydronaphthalen-2-yl)-N,N,2-trimethylprop-2-enamide (hereinafter "APX2032")). Additional suitable analogs are shown below and in Table 1. Further information on APX3330 may be found in Abe et al., U.S. Pat. No. 5,210,239, and information on APX2009 may be found in Kelley et al., J Pharmacol Exp Ther.2016 Nov, 359(2): 300-309, each incorporated herein by reference to the extent they are consistent herewith. 5 W4.5 83. 833. 873.2 74.7 53.0 14.0 54.M7 1 2 2 5 2 252 3 2) 2H e)e t)t2 3 3 C H 2 MEH H H M(E(CC(C O N N N N N N N 2 2 2 2 2 2 R R R R R R R2 2)R20X C= C= C= C= C= C= C= C=-7H H H H H H H H2C C C C C C C C0-41 323 3 30R HOeOeOeOeH H H2C M M M M C C C(92N0U00.:O D 03 60 70 80 90 01 1 251PI317TelM D 3 02 02 02 02 0 1 2 0 1 2 02 O XP XP XP XP XP XP XP XP92CbPaC T A A A A A A A A 6 4.353 3 O N72 H22 C O = enoniuq ohtpanO = 9 H4 C eeonily n lodMoh)t2eor epiPxerP -tHeO p)eproE(M Oc- M OE-c- iM(MN N N N N NP- 2 N - E R2R2R2R2R2R2 2 2 2)R R R20C= C= C= C= C= C= C= C= C=-7H H H H H H H H H2C C C C C C C C C0-423 3 3 3 3 3 3 30HOeH H H H H H H2C M C C C C C C C(920003 4 5 6 7.150 1 2 0 1 1 1 81 91 02 12 2 02 02 02 02 02 02 0217T XP XP XP XP XP X X X X92C P P P P P A A A A A A A A A 4 4.763 4 O N52 H22 C O = enoniuq ohtpanO = 9 H4 C onil o H H h C2 C p 2 roH He2 )t2)t2)eM 2 C C - M HN NOE(HE(M(Z N - E - Z N N O N N R2R2R2R2 2 2 2 2)R R R R R20C= C= C= C= C= C= C= C= C=-7H H H H H H H H H2C C C C C C C2C C0-423 3 3 30H H H HOe2C C C ClClHeC O M M(920002 3 4 5 6 7 8 9 0.250 2 2 0 2 2 0 2 2 0 2 2 0 2 2 2 3 2 02 02 02 0217T XP XP XP XP XP XP X X X92C P P P P A A A A A A A A A 9 2.582 4 O N51 H61 C O = enoniuq ohtpanO = 3 H C )2 23 3)3 2 2 2 2 H H H)C Ct)Et)Et)EtC(H H( ( ( (E(N N O O N N N N N 22 2 2 2 2 2 22)R R R R R R R R R20C= C= C= C= C= C= C= C= C=-7H H H H H H H H H2C C C C C C C C C0-4OeOeOeOeO33 3320 eF H H H2M M M M M C O C C C(920001 2 3 4 3 4 5 6 7.350 3 2 0 3 2 0 3 2 0 4 2 0 4 2 0 4 4 4 2 02 02 0217T XP XP XP XP XP XP XP X X92C P P P A A A A A A A A A 1 5.793 O = OeOeOeM M M e e e M O M O M O O = O = O = 919191H9 H9 H9 C C C62O C)O(C)O(C)O(C)O(3 H C C 3CH)3O)3 C H H OC(C(N N H N 2 2 2)R R R20C= C= C= -7H H H2C C C0-43 3 320H H H2C C C(920008.4 9 050 4 2 0 5 2 0217T X X X C P P P92P A A A Suitable dosages of the APE1 inhibitor, pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof, for use in the methods of the present disclosure will depend upon a number of factors including, for example, age and weight of an individual, severity of the UC symptoms, nature of a composition, route of administration and combinations thereof. Ultimately, a suitable dosage can be readily determined by one skilled in the art such as, for example, a physician, a veterinarian, a scientist, and other medical and research professionals. For example, one skilled in the art can begin with a low dosage that can be increased until reaching the desired treatment outcome or result. Alternatively, one skilled in the art can begin with a high dosage that can be decreased until reaching a minimum dosage needed to achieve the desired treatment outcome or result. Pharmaceutically acceptable salts are known to one skilled in the art and acceptable salts include salts of benzoquinone derivatives with inorganic acids, such as hydrochloride, hydrobromide, sulfate, and phosphate; those with organic acids, such as acetate, maleate, tartrate, methanesulfonate, benzenesulfonate, and toluenesulfonate; and those with amino acids, such as arginine, aspartic acid, and glutamic acid. Pharmaceutically acceptable salts are also in the form of metallic salts such as sodium (Na), potassium (K), calcium (Ca), and magnesium (Mg) salts, and these metallic salts as well are within the scope of the pharmacologically acceptable salts of the present disclosure. In one particularly suitable embodiment, the APE1 / Ref-1 inhibitor is APX3330, and the subject is administered from about 1.0 µM to about 50 µM APX3330. In another embodiment, the subject is a human patient and is the APE1 / Ref-1 inhibitor is administered in vivo in amounts of from about 10 mg / day to about 600 mg / day. In some embodiments, the APE1 inhibitor is administered via a composition that includes the APE1 inhibitor and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may be, for example, excipients, vehicles, diluents, and combinations thereof. For example, where the compositions are to be administered orally, they may be formulated as tablets, capsules, granules, powders, or syrups; or for parenteral administration, they may be formulated as injections (intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intravitreal), drop infusion preparations, or suppositories. These compositions can be prepared by conventional means, and, if desired, the active compound (e.g., APX3330) may be mixed with any conventional additive, such as an excipient, a binder, a disintegrating agent, a lubricant, a corrigent, a solubilizing agent, a suspension aid, an emulsifying agent, a coating agent, or combinations thereof. It should be understood that the pharmaceutical compositions of the present disclosure can further include additional known therapeutic agents, drugs, modifications of the synthetic compounds into prodrugs, and the like for alleviating, mediating, preventing, and treating the diseases, disorders, and conditions described herein. For example, in one embodiment, the APE1 inhibitor can be administered with one or more of current therapeutic agents and drugs for treating UCC (e.g., echinomycin, 5-aminosalicylic acid (5-ASA), corticosteroids, azathioprine, 6-mercaptopurine, methotrexate, cyclosporine, tacrolimus, anti-TNF drugs (e.g., infliximab, certolizumab, adalimumab, and golimumab), vedolizumab, natalizumab, ustekinumab, probiotics, antibiotics, anti-inflammatories (e.g., mesalamine (Asacol HD, Delzicol, others), balsalazide (Colazal) and olsalazine (Dipentum), and the like). In one particular embodiment, the APE1 inhibitor is administered with the HIF-1α inhibitor, echinomycin. As discussed below, it was shown that the HIF-1α inhibitor, echinomycin, inhibited IL- 1r1 signaling. HSCs from chronic UC mice bearing Dnmt3a- / -mutation were functionally impaired and differentiated into more myeloid-biased HPC-2 progenitors and fewer MEPs under the transplant settings, indicating that chronic UC contributes to the manifestation of age-associated HSC phenotypes in Dnmt3a- / -mice The pharmaceutical compositions including the APE1 inhibitor, alone or in a combination therapy, and / or pharmaceutical carriers used in the methods of the present disclosure can be administered to a subset of individuals / subjects in need. As used herein, a "subject in need" refers to an individual at risk for or having UC. As such, in some embodiments, the methods disclosed herein are directed to a subset of the general population such that, in these embodiments, not all of the general population may benefit from the methods. Based on the foregoing, because some of the method embodiments of the present disclosure are directed to specific subsets or subclasses of identified individuals (that is, the subset or subclass of subjects "in need" of assistance in addressing one or more specific conditions noted herein), not all individuals will fall within the subset or subclass of individuals as described herein. In particular, the individual in need is a human. The individual in need can also be, for example, a research animal such as, for example, a non-human primate, a mouse, a rat, a rabbit, a cow, a pig, and other types of research animals known to those skilled in the art. Various functions and advantages of these and other embodiments of the present disclosure will be more fully understood from the examples shown below. The examples are intended to illustrate the benefits of the present disclosure, but do not exemplify the full scope of the disclosure. EXAMPLE In this Example, the APE1 / Ref-1 / HIF-1α / IL-1r1 signaling pathway was analyzed for its role in aberrant hematopoiesis and the APE-1 / Ref-1 / HIF-1α / IL-1r1 signaling was analyzed for its ability to serve as a potential therapeutic target for chronic UC. Materials and Methods Mice 8–10-week-old C57BL / 6 (B6) male mice, procured from the In Vivo Core facility at Indiana University, were housed in pathogen-free standard laboratory conditions (22±1°C Temp, 12:12-h light / dark cycle) at the Indiana University School of Medicine. All mice were given ad libitum access to a normal chow diet, and water throughout the study. All animal procedures were conducted in accordance with the Guidelines for the Care and Use of Laboratory Animals and protocols approved by the Institutional Animal Care and Use Committee at Indiana University School of Medicine. Chronic DSS-induced Ulcerative Colitis model The Chronic Ulcerative Colitis (UC) model was developed by oral administration of DSS (MW 36-50 kDa colitis grade, MP Biomedicals, Santa Ana, CA). After acclimation, mice were randomly assigned to two groups (n=5 per group): For the chronic DSS-induced UC, mice were treated with 4 cycles of DSS for 5 days and 7 days of resting (drinking water) between each cycle. The day DSS started was considered day 0. Body weight, stool softness, and blood in the rectum or stool were recorded daily during DSS and recovery cycles. After the 4th cycle of DSS treatment, mice were sacrificed, and tissue samples were harvested for analysis (FIGS.1A-1G). Assessment of Ulcerative Colitis (UC) severity To evaluate the progression of ulcerative colitis, a disease activity index (DAI) was assessed during the DSS treatment cycles as described in Horuluoglu, B.H., et al., PAM3 protects against DSS-induced colitis by altering the M2:M1ratio. Sci Rep, 2020.10(1): p.6078. The DAI is an aggregate score determined by factors such as body weight loss, stool softness, and blood in the rectum or stool. Body weight loss was scored as follows: score 0, no weight loss compared to initial weight; score 1, weight loss within 1-5%; score 2, weight loss within 5-10%; score 3, weight loss within 10-20%; and score 4; weight loss >20%. Stool consistency score was determined as follows: score 0, normal (solid pellet); score 1, soft but in pellet shape; score 2, loose stool but with some solidity; score 3, loose stool with sign of liquid consistency; and score 4, watery diarrhea. Rectal bleeding was scored as follows: score 0, no sign of blood, score 1, no bleeding; score 2, slight bleeding; score 3, bloody diarrhea; and score 4, gross bleeding. Assessment of progression of myelopoiesis The progression of myelopoiesis in mice was assessed in resting phase after each DSS treatment cycle by counting white blood cells (WBCs) using Element HT5-Heska and flow-cytometry analysis of peripheral blood stained with Mac1, Gr1 and B220 antibodies. The progression of myelopoiesis in mice was assessed in resting phase after each DSS treatment cycle by counting white blood cells (WBCs) using Element HT5-Heska and flow-cytometry analysis of peripheral blood stained with Mac1, Gr1 and B220 antibodies. Colon lengths measurement Colon lengths were measured on sacrifice day from the ileocecal junction to the rectum and the unit expressed in centimeters (cm). Isolation of lymphocytes from the lamina propria (LP) of the colon Colons were opened longitudinally and washed gently with PBS. Then colons were excised into small pieces and incubated at 37 °C in RPMI 1640 (Lonza) supplemented with 5% heat-inactivated fetal bovine serum (FBS), 100 U / ml penicillin, 100 mg / ml streptomycin, 25 nmol / L HEPES, 2mM L-glutamate, 55mM 2-ME and 2mM EDTA (to remove epithelial cells). After three 20 min of washing to remove ETDA, tissue was transferred to complete RPMI 1640 containing 1 mg / ml Collagenase IV (Sigma) and 200 U / ml DNase I (Sigma) and incubated at 37 °C for 1hr. The suspension was passaged through 70 μM nylon filter, centrifuged at 450×g for 10 min at 4°C. Then, cells pellet washed one more time with ice-cold complete RPMI 1640 to remove Collagenase. LP lymphocytes were enriched on a 40 / 70% percoll (GE Healthcare) gradient centrifugation for 20 min at 750×g speed at 21°C without acceleration and brake

[0033] , and were used for subsequent analysis. Immunophenotyping Immunophenotyping of cells was executed as described in Ghosh, J., et al., S6K1 regulates hematopoietic stem cell self-renewal and leukemia maintenance. J Clin Invest, 2016.126(7): p.2621-5. A single cell suspension of bone marrow and spleen was prepared. Briefly, bone-marrows were flushed with Iscove Modified Dulbecco Medium (IMDM, Invitrogen). For making single cell splenocytes, spleens were crushed between the microscopic slides, flushed using syringe filled with IMDM and filtered through a 50 μm nylon filters. Red blood cells (RBCs) from bone-marrow and splenocytes were lysed in a 0.8% NH4Cl RBC lysis buffer for 5 min at room temperature and then cells were resuspended in phosphate buffered saline containing 1% bovine serum albumin (BSA, Sigma) and 10% rat serum (Sigma). After counting the cells in cell viability analyzer (Vi-CELL-XR, Bechmann Coulter), equal number of cells were used for staining with flow antibodies. Intracellular flow cytometry (ICFC) was performed as per Cai, Z., et al., Inhibition of Inflammatory Signaling in Tet2 Mutant Preleukemic Cells Mitigates Stress-Induced Abnormalities and Clonal Hematopoiesis. Cell Stem Cell, 2018. 23(6): p.833-849 e5. Briefly, freshly prepared bone marrow cells and colonic LPL cells were pre-stained by using cell surface antibodies for HSPCs or for mature cells and then fixed with BioLegend Cytofix and washed using BioLegend Cytoperm three times. Then, pre-stained cells were re-stained with the appropriate indicated antibodies of intracellular proteins. Staining with an Annexin-V and 7-AAD kit (BioLegend, Cat # 640922) was performed according to the manufacturer’s instruction for apoptosis analysis, along with labeling of LSK cells. A list of flow antibodies is provided in Table 2. Table 2 Reagents Company Catlog No. 29715.00029 (2024-027-02) PCT CD150 PerCPCy5.5 Biolegend Cat # 115922CD34 FITC eBiosciences Cat #11-0341-82g 29715.00029 (2024-027-02) PCT p-NF-kB p65 (Ser536) AF647 Cell Signaling Cat # 4887SRat IgG1, k Isotype BV785 Biolegend Cat # 400443 Serum cytokine profiling Serum was separated from peripheral blood and submitted to Eve Technologies (Canada) for serum cytokine analysis. Results are expressed as pg / mL or as ng / mL of serum. Competitive repopulation assay For competitive bone marrow transplantation (cBMT) assay, recipient (F1 mice, CD45.1+ / CD45.2+) mice were irradiated at 1100 cGy from a cesium source using a split two dose of 700cGy and 400 cGy at an interval of 4 hrs.0.2×106BM mono-nuclear donor cells from either B6-control or B6-mice treated with 3% DSS (w / v) (CD45.2+) along with 0.2×106 Boy / J (CD45.1+). BM mono-nuclear competitor cells were transplanted by intravenous injection into lethally irradiated F1 recipient mice (n=5 per group). Engraftment of donor cells (B6-control and B6-mice treated with DSS) was determined at an interval of 4 weeks by flow cytometry analysis of peripheral blood. At 16th week post transplantation, mice were sacrificed for complete analysis (a schematic presentation is provided in FIG.2). In another cBMT assay where 0.2×106BM mono- nuclear donor CD45.2+cells from either Veh, DSS+Veh or APX3330+3% DSS treated B6-mice used along with 0.2×106Boy / J (CD45.1+) BM mono-nuclear competitor cells were transplanted by intravenous injection into lethally irradiated F1 recipient mice (n=5 per group). Engraftment of donor cells was determined as above, and mice were sacrificed for complete analysis at 20th week post transplantation (a schematic presentation is provided in FIG.3). In vivo Ref-1 / APE-1 inhibitor APX3330 and HIF-1α specific inhibitor Echinomycin drug treatment Twenty, 8-10 weeks old male B6-mice were randomized into 4 groups: vehicle (Veh) control, DSS+Veh, and APX3330+DSS group (n=5 mice per group). APX3330 drug, a kind gift from Dr. Mark R. Kelley at Department of Pediatrics, Indiana University School of Medicine, Indianapolis, was dissolved in Cremophor: Ethanol (1:1) (Cremophor from Sigma, catalog # C5135) for stock solution preparation and diluted with 0.5% methylcellulose prior to use. Mice were treated either with vehicle or APX3330 drug at 50 mg / kg body weight, twice a day by oral gavage, one week before the start of DSS treatment and continued throughout the 4 cycles of 3% (w / v) DSS treatment. Similarly, mice were treated either with DMSO-vehicle or Echinomycin at 10 μg / kg body weight on alternate days through intraperitoneal injection (i.p.). The progression of disease was monitored as above. At the end of the last resting cycle, mice were sacrificed, and tissues were harvested for analysis. Statistical analyses All data were presented as mean ±SD. Statistical significance between groups was analyzed using GraphPad Prism software version 7.0 (GraphPad, San Diego, CA). For the analyses, the statistical significance was assessed by either one-way analysis of variance (ANOVA) with Tukey’s multiple comparison test, two-way ANOVA with Sidak’s multiple comparison test or an unpaired, Student’s two-tailed t-test. P values <0.05 were considered statistically significant. Results APE-1 / Ref-1 inhibitor, APX3330 rescues chronic UC mediated hematopoietic defects. Inflammatory signalings arise from the functional interactions between gut microbiota and the host immune system and may likely contribute to disease pathophysiology, not only in the intestine, but also in distant organs, including the lungs, liver, and the BM. Oxidative stress is a key factor intimately associated with the development and course of inflammation in UC. In this context, apurinic / apyrimidinic endonuclease 1 / reduction oxidation factor-1 (APE1 / Ref-1; also called Ref-1) protein regulates oxidative stress by modulating reduction-oxidation (redox)-activities of various transcription factors which are known to be involved in the cell survival and inflammation. It was hypothesized that chronic inflammatory conditions, like UC, would contribute to the abnormal hematopoiesis in mice, and the blockade of redox activity of the APE1 / Ref-1 protein by the specific inhibitor, APX3330, would reverse chronic UC- induced hematopoietic defects. The various subsets of hematopoietic stem cell (HSC) were analyzed as shown in the FIG.1H. The frequency of long-term hematopoietic stem cells (LT-HSCs; LSKgatedCD48-CD150+) (Fig.1D & FIGS.1I & 1J) and multipotent progenitors (MPPs; LSKgatedCD48-CD150-); also known as short-term hematopoietic stem cells (ST-HSCs) (Fig.1E & FIGS.1K & 1L) was reduced; however, the frequency of myeloid-biased hematopoietic progenitor cell-2 (HPC-2; LSKgatedCD48+CD150+) (Fig.1F & FIGS.1M & 1N) was elevated in the BM of DSS-treated mice compared to non-DSS treated control mice. The absolute numbers of various hematopoietic cells (LSKs, LT-HSCs, and HPC- 2) (FIGS.4A-4F) were increased in the BM of DSS-treated mice compared to control mice. Chronic UC mice treated with the ref-1 inhibitor APX3330 showed a significant reduction in the absolute number of bone marrow cells compared to control chronic UC mice (FIG.1C). Likewise, the ref-1 inhibitor, APX3330, corrected the frequencies of LSKs, LT-HSCs, MPPs and myeloid-biased HPC-2 cells in chronic UC mice to the level of veh-treated control mice (FIGS.1D, 1H, and 1M). Along these same lines, absolute numbers of LSKs, LT-HSCs and HPC-2 cells were also restored in the BM of chronic UC mice after APX3330 treatment to the levels seen in veh-treated mice (FIGS.4D-4F). Since it was observed that chronic UC mice showed splenomegaly characterized by increased spleen weight and elevated spleen cellularity (FIGS.1O-1T), it was suspected that splenomegaly in chronic UC mice might be due to the “extra- medullary hematopoiesis”. Therefore, the spleens of mice were analyzed, and as expected, an expansion in the frequencies of LSKs (FIGS.1U & 1V), LT-HSCs (FIG. 1W), MPPs (FIG.1X), and myeloid-biased HPC2 (FIG.1Y) were found in the spleen of chronic UC mice compared to control mice. Relative to control mice, absolute numbers of LSKs, LT-HSCs, MPPs, and HPC-2 cells were elevated in the spleen of DSS-treated mice (FIGS.4G-4N). These data suggest that chronic UC can lead to extramedullary hematopoiesis in the spleen. Then, it was investigated if ref-1 inhibitor, APX3330, corrected spleen abnormalities observed in chronic UC mice. It was found that APX3330 treatment also restored the spleen weight, spleen cellularity (FIGS.1O-1Q), frequency of LSKs (FIG.1U), and absolute numbers of various hematopoietic cells (FIGS.4K-4N). Downstream of HSCs and MPPs, c-Kit+ hematopoietic progenitors were examined and found that the frequencies of LKs (Lin-c-Kit+) (FIG.1Z, and FIGS.5A left panel & 5B) and granulocyte-monocyte progenitors (GMPs; LKgatedCD16 / 32hiCD34+) (FIG.1Z, and FIG.5C) were significantly increased; whereas, the frequencies of common-myeloid progenitors (CMPs; LKgatedCD16 / 32lowCD34+) (FIG.1BB and FIG.5D), megakaryocyte-erythroid progenitors (MEPs; LKgatedCD16 / 32-CD34-) (FIG.1CC and FIG.5E) and common-lymphoid progenitors (CLPs; Lin-CD127+c-KitloSca-1lo) (FIG. 5F) were significantly declined in the BM of chronic UC mice. Likewise, the absolute numbers of LKs and GMPs (FIGS.5G & 5P-5Q) were increased, and those of CMPs and CLPs (FIGS.5I-J and FIG.5R) were decreased in the BM of chronic UC mice as compared to control mice. Relative to control mice, the spleens of chronic UC mice revealed an increase in the frequencies of LKs (FIG.1DD and FIG.5K) and GMPs (FIG. 1EE and FIG.5L) as well as in the absolute numbers of LKs, GMPs, and CMPs (FIGS. 5M-5O & FIGS.5S & 5T). Although chronic UC mice treated with ref-1 inhibitor, APX3330 did not show any corrections in the frequency of LKs and CMPs (FIGS.1Z & 1BB); however, the frequencies of GMPs (FIG.1AA), and MEPs (FIG.1CC) were restored in the BM to levels observed in veh-control mice. Similarly, the treatment with the ref-1 inhibitor, APX3330, normalized the frequencies of LKs (FIG.1DD), GMPs (FIG.1EE) and MEPs (FIG.1FF); and the absolute numbers of LKs and GMPs (FIGS. 5S & 5T) in the spleens of chronic UC mice as compared to veh-control mice. Given the changes in the frequency and absolute numbers of HSC / HPSCs within the LSK pool of chronic UC, it was assessed if chronic inflammation of UC contributes to HSC / HSPC survival and / or differentiation and apoptosis. Apoptosis in the LSK pool was analyzed by Annexin-V and 7-AAD staining, followed by flow-cytometry analysis. A remarkable (>2 fold) decrease in the apoptosis of LSK cells was found from chronic UC mice compared to control mice (FIGS.6A & 6B). In a subsequent analysis of cell cycle in LSK cells by DAPI (4’,6-diamidino-2-phenylindole) staining followed by flow-cytometry, it was noticed that LSKs have increased cell cycle at G0 / G1 and S-phase (FIGS.6C & 6D) in chronic UC mice, suggesting that chronic UC contributes to both enhanced survival and increased cell cycle progression in LSK cells compared to controls. In the subsequent experiment, it was determined if the hematopoietic defects observed in the BM and spleen of chronic UC mice were indeed due to chronic UC. DSS-induced colitis model is characterized by ulceration and submucosal inflammation provoked by disruption of epithelial barrier and exposure to luminal microbiota. Circulating monocytes are innate immune cells that play an important role in colon tissue homeostasis both in humans with UC and experimental model of colitis. These cells can be categorized into two subsets: patrolling monocytes (identified as CD11b+, CX3CR1lo, Ly6Clo, CCR2-, and CD115+) and inflammatory monocytes (identified as CD11b+, CX3CR1lo, Ly6Chi, CCR2+, and CD115+). Ly6Chi monocytes are generated from common monocyte progenitors (cMoPs) in the BM and released into circulation in a CC-chemokine receptor-2 (CCR2) dependent manner to infiltrate the affected tissue. As Ly6Chi monocytes are conventionally linked to inflammation and Ly6Clo monocytes with anti-inflammation, it was explored whether these subsets of monocytes were altered in the BM and spleen of mice under chronic UC. It was found that the frequency of Ly6Chi monocytes was significantly reduced and the frequency of Ly6Clo monocytes was increased in the BM (FIG.1GG), while the frequency of Ly6Chi monocytes was significantly elevated and the frequency of Ly6Clo monocytes was decreased in the spleen of chronic UC mice (FIG.1HH). The reduction in the frequency of Ly6Chi monocytes indicates the emigration of Ly6Chi monocytes from the BM to the spleen and inflamed colon. Surprisingly, APX3330 treatment inhibited the frequency of proinflammatory Ly6Chi monocytes and supported the frequency of anti-inflammatory Ly6Clo monocytes in chronic UC mice. Furthermore, in accordance with earlier reports, the mice treated with 3% DSS exhibited several features of chronic UC, including transient body weight loss (FIG.7A & 6E) and an increase in the disease activity index (DAI) (FIG.7B & 6F) which is an aggregate score determined by body weight loss, stool softness, and blood in the rectum or stool and shortening of the colon length (FIGS.7C & 7D FIGS.6G-6I). It was also found that mice with chronic UC showed a significant accumulation of pro-inflammatory cells such as, myeloid cells (FIG.7E and FIGS.6J- 6M), B220+cells (FIG.6N) and T cells (FIG.7F and FIGS.6O & 6P) in the colon. Since mice lacking regulatory T (Treg) cells develop more spontaneous colitis, Treg cells were analyzed in the colon lamina propria (cLP) of colons and found that the frequency of Tregs was severely reduced in the colon of mice with chronic UC as compared to control mice (FIG.6Q). Then, it was assessed if the administration of APX3330 would also correct pathophysiological conditions associated with defective gut. Interestingly, mice treated with APX3330 reduced body weight loss and disease activity index, and improved colon size (FIGS.7A-7D). Surprisingly, APX3330 completely reversed colon size close to the length of control groups treated either with veh or APX3330 alone (FIG.7C and FIG. 8D). Remarkably, the accumulation of proinflammatory myeloid (CD11b+Gr-1+) (FIG. 7E) and CD3+T cells (FIG.7F) was also highly reduced in the colon of chronic UC mice upon APX330 administration. Additionally, it was found that mature cell populations such as CD11b+Gr-1+neutrophils, B220+B cells, and CD3+T cells which were altered in the PB (FIGS.7G-7I), BM (FIGS.7J-7L) and spleen (FIGS.7M-7O) of UC mice, were also normalized upon APX3330 treatment. These data indicate that chronic inflammatory condition in UC drives the expansion of HSPCs and their differentiation toward myelopoiesis in a feed-forward loop and the blockade of redox activity of the ref-1 by its specific inhibitor, APX3330 restores chronic UC-induced hematopoietic defects to the normalcy. APE-1 / Ref-1 inhibitor APX3330 rescued chronic UC associated phenotypes via inhibition of G-CSF, chemokine KC and hypoxia-inducible transcription factor-1α (HIF-1α). In a subsequent analysis, a multiplex serum cytokine profiling from mice treated with veh or APX3330 and with / or without 3% DSS was performed. The expression of 31 cytokines and chemokines covering a broad spectrum of immune and inflammatory mechanisms was measured. The expression of 31 cytokines and chemokines covering a wide spectrum of immune and inflammatory mechanisms was measured. Among the 31 cytokines / chemokines, levels of 11 cytokines or chemokines (G-CSF, IL-1β, IL-6, IL-10, IP-10, IL-17, MCP-1, MIG, MIP-1α, MIP-1β, and TNF-α) were found to be significantly elevated (FIG.9A). Inflammatory signals, like IL-1β, IL-6, IL-17, and TNF-α have been implicated in accelerating proliferation and myeloid differentiation of human and murine HSPCs, were elevated in the serum of chronic UC mice. Notably, the level of the granulocyte-colony stimulating factor (G-CSF) was the most abundant in chronic UC compared to control mice (FIG.8A and FIG.9A). The high level of G-CSF expression has been shown in the serum of mice during systemic infection, which induces mobilization of HSPCs in the BM by repressing the CXCL12 / CXCR4 signaling axis in osteoblasts. Moreover, the high level of expression of chemokines was found responsible for recruiting neutrophils and monocytes / macrophages such as keratinocyte chemoattractant KC (CXCL1) (FIG.8A), IP-10 (CXCL10), MCP-1 (CCL2), MIG (CXCL9), MIP-1α (CCL3), and MIP-1β (CCL4) (FIG.9A) in the serum of chronic UC mice. Consistent with the corrections in the hematopoietic defects in chronic UC mice, APE-1 / Ref-1 inhibitor APX3330 also reduced the expression of G-CSF, and the chemokine KC induced by a chronic inflammatory condition of UC (FIG.10A). Since APE-1 / Ref-1 regulates redox-sensitive DNA binding activity of transcription factor NF-ĸB, and given that NF-ĸB is the central mediator of the inflammatory response, it was next analyzed if chronic UC impacted the expression of NF-ĸB in HS / PCs. Intracellular staining of phospho-NF-ĸBp65 was performed, followed by flow cytometry analysis. There was no detection of NF-ĸB activation in LK cells in the BM (FIGS.8B & 8C) or in the spleen of chronic UC mice (data not shown). Having seen elevated serum levels of G-CSF in chronic UC mice and, given the known role for G-CSF in stabilizing and activating hypoxia-inducible factor-1 α (HIF-1α), even under normoxic conditions, it was hypothesized that G-CSF secreted in chronic UC would activate HIF-1α in HSPC, and the ref-1 inhibitor, APX3330, would inhibit G-CSF- mediated activation of HIF-1α in these cells. Using HIF-1α intracellular staining followed by flow-cytometry, it was found that chronic UC mice showed highly enriched frequency as well as the absolute number of HIF-1α+LK cells in the BM (FIGS.8D, 8E and 8G) and in the spleen (FIGS.8F & 8H). As expected, the frequency and the absolute number of HIF-1α+LK cells in the BM (FIGS.8D, 8E and 8G) and in the spleen (FIGS.8F & 8H) of chronic UC mice were normalized upon APX3330 treatment. Collectively, the data show that chronic UC specifically induces HIF-1α expression in HS / PCs via APE- 1 / Ref-1 / G-CSF activation and drives differentiation of HSPCs toward granulopoiesis, whereas; the ref-1 inhibitor, APX3330, curtails the expression of HIF-1α by inhibiting G- CSF induction at the primitive hematopoietic progenitor cell level and resolves chronic UC. The blockade of redox activity of APE-1 / Ref-1 by its inhibitor, APX3330 improves repopulating ability of chronic UC triggered defective HSC under transplant setting. Given that under transplant setting, HSCs derived from chronic UC mice tendered reduced repopulating ability (FIGS.9C-9E) and that ref-1 inhibitor, APX3330, treatment rescued chronic UC-induced defective hematopoiesis, it was interrogated if APX3330 treatment would also restore HSC function in chronic UC mice. A competitive repopulation assay was performed using whole BM cells derived from veh control, chronic UC (DSS+veh), and chronic UC mice treated with APX3330 (APX3330+DSS), as described in the APX3330 treatment protocol (Materials and Methods). A schematic model for conducting competitive BM transplantation (cBMT) is shown in FIG.10A. In agreement with the previous observation (FIGS.9C-9E), reduced PB chimerism of CD45.2+donor cells derived from chronic UC mice was consistently observed as compared to veh treated control mice; whereas, CD45.2+donor cells obtained from chronic UC mice treated with APX3330 showed improved repopulating ability compared to chronic UC mice (FIG.10B). At 20 weeks post-transplantation, mice were sacrificed for analysis of chimerism, including for the presence of HSCs / HSPCs and mature cells in the BM. Consistent with the restoration of PB chimerism, CD45.2+donor cells derived from chronic UC mice treated with APX3330 showed a significantly improved repopulating ability in the BM at the comparable level of the veh group (FIG.10C). Under cBMT setting, F1 recipient mice transplanted with CD45.2+ donor cells derived from chronic UC mice treated with APX3330, showed complete restoration of the BM cellularity (FIG.10D), and more interestingly within the CD45.2+donor-derived cells, the frequencies of LSKs (FIG.10E), LT-HSCs (FIG.10F), MPPs (FIG.10G), and myeloid-biased HPC-2 cells (FIG.10H) were restored in the BM to levels seen in F1 recipient mice who received CD45.2+donor cells of the veh control group. Likewise, within the CD45.2+donor-derived cells, downstream of HSCs and MPPs, the frequencies of LKs (FIG.10I), GMPs (FIG.10J), MEPs (FIG.10K), and CD11b+Gr-1+myeloid cells (FIG.10L) were also rescued. To interrogate if donor derived CD45.2+cells from chronic UC had any effects on the colon, the respective colons in FIG.10M were quantified and found that recipient mice transplanted with CD45.2+donor-derived cells of chronic UC displayed reduced colon size, and interestingly, the colon size of recipient mice who received CD45.2+donor-derived cells of APX3330 treated chronic UC mice was restored to the comparable level in veh-treated control mice (FIGS.10M & 10N). This data indicates that APX3330 treatment rescued the gut in a hematopoietic stem cell autonomous manner. Collectively, the data under transplantation conditions show that ref-1 mediates its effect via regulating G-CSF secretion and HIF-1α activity, and the blockading of the redox activity of ref-1 by APX3330 improves HSCs function and corrects pathophysiological conditions related with chronic UC. HIF-1α specific inhibitor, echinomycin reduces accumulation of proinflammatory CD4+T helper 1 (Th1) and Th17 cells in the colon by reducing IL-1β and IL-6 production. Having established that the ref-1 inhibitor, APX3330, suppressed the expression of HIF-1α in HSPCs and resolved chronic UC, it was aimed to understand the role of HIF-1α signaling downstream of APE-1 / Ref-1 in chronic UC by utilizing a HIF- 1α specific inhibitor, echinomycin. To this end, a cluster of mice were divided into 3 groups: veh, DSS+veh, and DSS+Echin. DSS+Echin group was treated with HIF-1α inhibitor, echinomycin, at 10 µg / kg body weight by i.p. injection on alternate days; whereas, the veh and DSS+veh group received DMSO-vehicle (veh), a week before the DSS treatment depicted as in FIG.1B. Similar to the data observed with APX3330 administration, the echinomycin treatment resulted in improved body weight, reduced DAI score, and correction in the colon size of chronic UC mice (FIGS.11A-11C). The echinomycin treatment was also found to reduce splenomegaly and the BM cellularity to levels seen in veh-treated mice (FIGS.11D-11F). Immune cell infiltration of the intestinal mucosa is critical for both the development and maintenance of UC in patients and mice. Given that the APX3330 treatment suppressed the elevated frequency of Ly6Chi monocytes in the spleens of chronic UC mice (FIG.7Q), and the recruitment of Ly6Chi monocytes to the inflamed tissues is known to regulate T cell responses, including differentiation of pathogenic Th1 / Th17 cells, it was examined whether the echinomycin would revert the mucosal infiltration of CD4+T cells as well as their differentiation into colitogenic CD4+T helper cells in the colon of chronic UC mice. The treatment of echinomycin normalized the frequencies of CD4+T cells, Treg cells, colitogenic Th1 and Th17 cells, and the ratio of Treg / CD4+T cells in the colon of chronic UC mice (FIGS.11G-11K). Because the expression of IL-1β, IL-6 and TNF-α was enhanced in the serum of chronic UC mice (FIG.9A), and given that these cytokines have established role in the development and differentiation of colitogenic Th1 and Th17 from naïve CD4+T cells, IL-1β, IL-6 and TNF-α expression was intracellularly analyzed at the cellular level in cLP lymphocytes. Although the expression of TNF-α was unaltered, the expression of IL-1β and IL-6 was found at significantly higher levels in CD11b+myeloid cells and CD3+T cells within the CD45+leukocytes, which were reduced by echinomycin treatment (FIGS.11L & 11M). Various studies have shown that IL-1β regulates the production of G-CSF and GM-CSF in Th1 and Th17 cells and plays a significant role in autoimmune EAE, colitis, and liver cancers. cLP Th1 cells were found to produce significantly higher levels of GM-CSF (3.25-fold), while Th17 cells produced both G-CSF (1.92-fold) and GM-CSF (4.07-fold) in chronic UC mice, which were reduced by echinomycin treatment (FIGS.11N-11P). Overall, the data indicate that HIF-1α downstream of APE1 / Ref-1 regulates chronic UC inflammation via IL-1β; whereas, the blockade of HIF-1α by echinomycin attenuate chronic UC by reducing proinflammatory Ly6Chi monocytes and suppressing colitogenic Th1 and Th17 cells. HIF-1α specific inhibitor, Echinomycin, restores chronic UC induced defective hematopoiesis. Since the expression of G-CSF and GM-CSF by Th1 and Th17 cells in chronic UC mice was normalized by echinomycin (FIGS.11N-11P), and high levels of G-CSF and GM-CSF contribute to dysregulated hematopoiesis, it was further assessed if the blockade of HIF-1α by echinomycin would correct defective hematopoiesis observed in chronic UC mice. Like the previous results with APX3330 treatment, the BM cellularity (FIG.12A) and the frequency of LSKs (FIG.12B) was reduced; whereas, the frequencies of LT-HSCs (FIG.12C) and MPPs (FIG.12D) were improved in the BM of chronic UC mice by the treatment of echinomycin. The myeloid-biased HPC-2 progenitors that appear to be immediately downstream of MPPs at early stages of lineage restriction, give rise to myeloid cells were also corrected by the echinomycin treatment (FIG.12E). Furthermore, the absolute numbers of LSKs, LT-HSCs, MPPs, and HPC-2 cells (FIGS.13A-13D) were reversed by the echinomycin treatment. Similarly, the frequencies of LSKs, LT-HSCs, and MPPs (FIGS.12F-12H) and absolute numbers of LSKs, LT-HSCs, MPPs, and myeloid-biased HPC-2 (FIGS.13E & 13H) were corrected by the treatment with echinomycin. Since the APX3330 treatment restored various lineage-committed hematopoietic progenitors (HSPCs), it was proposed that the echinomycin treatment would rescue abnormal c-Kit+HSPC subsets in chronic UC mice. As expected, the frequencies and absolute numbers of LKs GMPs, CMPs, MEPs, and CLPs (FIGS.12I - 12M and FIGS.13I-13M) in the BM of chronic UC were returned to normalcy by echinomycin treatment. Likewise, the frequencies and absolute numbers of LKs GMPs, CMPs, and MEPs (FIGS.12N-12Q and FIGS.13N-13Q) in the spleen of chronic UC were rescued by echinomycin treatment. Proportions of CD11b+Gr-1+neutrophils, B220+B cells, and CD3+T cells in PB (FIGS.12R-12T), in the BM (FIGS.12U-12W), and spleen (FIGS.12X-12Z) were significantly reversed by echinomycin treatment. These data suggest that echinomycin treatment rescues chronic UC driven aberrant HSCs and HSPCs by balancing myelopoiesis and lymphopoiesis. HIF-1α specific inhibitor, Echinomycin restores chronic UC induced defective hematopoiesis by regulating Interleukin-1receptor1 (IL-1r1) signaling in HSPCs. Since several investigators have demonstrated that IL-1r1signaling plays a critical role in HSPC cell differentiation and given that serum level of proinflammatory cytokine IL-1β was elevated in chronic UC mice, it was further hypothesized that the elevated expression of HIF-1α in HSPCs regulates IL-1r1 signaling under chronic UC conditions. Similar to the observation with APX3330, the expression of HIF-1α in immature LKs and CD34+HSPCs was reduced in the BM of chronic UC mice upon administration of echinomycin (FIGS.14A & 14B). When the immature LKs and CD34+ HSPCs were evaluated for the surface expression of IL-1r1, it was observed that IL-1r1 expression was heightened in immature LKs and CD34+HSPCs from BM of chronic UC mice, while the blockade of HIF-1α by echinomycin reduced the expression of IL-1r1 in chronic UC mice (FIGS.14C & 14D). This data indicates that HIF-1α promulgates its inflammatory effects through IL-1r1 signaling, and in concordance with the results, a previous study demonstrated that HIF-1α via its transcriptional target G-protein estrogen receptor (GPER) mediates feed-forward loop coupling IL-1β to the receptor IL-1r1 for driving inflammatory and invasive biological responses (Lappano, R., et al., The IL1beta- IL1R signaling is involved in the stimulatory effects triggered by hypoxia in breast cancer cells and cancer-associated fibroblasts (CAFs). J Exp Clin Cancer Res, 2020. 39(1): p.153). IL-1r1 signaling propagates upon binding ligands IL-1α, IL-1β, and IL-1 receptor antagonist (IL-1Ra) to the receptor IL-1R1

[0079] . The IL-1β / IL-1R1 signaling axis plays a critical role in the transmigration of inflammatory monocytes to various tissues. As the data showed IL-1R1 expression was reduced by echinomycin treatment that was elevated in HSPCs under the chronic UC settings, and other investigators have shown that loss of IL-1 signaling in IL-1R1 null mice globally impaired leukocytes recruitment reducing the number of infiltrating Ly6Chi and Ly6Clo monocytes in the infarcted myocardium, it was anticipated that administration of echinomycin would also suppress the proinflammatory Ly6Chi monocytes in the chronic UC mice. In the same lines with the previous data with APX3330 treatment (FIGS.7P-7Q), it was found that the frequency of Ly6Chi monocytes was significantly reduced and the frequency of Ly6Clo monocytes was increased in the BM (FIG.14E left & right panels) of chronic UC mice. This data indicates that Ly6Chi monocytes have been released in the circulation. Therefore, the PB and spleens of chronic UC mice were also analyzed for Ly6Chi and Ly6Clo monocytes and found that the frequency of Ly6Chi monocytes was remarkably elevated; whereas, the frequency of anti-inflammatory Ly6Clo monocytes was reduced in both the PB and spleens of chronic UC mice (FIGS.14F & 14G). Notwithstanding, these cells were restored by echinomycin to their normalcy in the BM, PB, and spleens of chronic UC mice (FIGS.14E-14G). Overall, the data revealed that HIF-1α mediates its inflammatory signaling via the IL-1r1 pathway that leads to abnormal differentiation of HSPCs, and the blockade of HIF-1α by echinomycin alleviates the inflammatory IL-1r1 pathway under chronic UC conditions. Chronic UC drives expansion of myelopoiesis, suppresses erythropoiesis, and profoundly impairs stem cell functions in pre-leukemic Dnmt3a mutant mice. Additionally, the impacts of chronic UC in clonal hematopoiesis of indeterminate potential (CHIP) were investigated. CHIP is characterized as clonal hematopoiesis with recurrent mutations in epigenetic regulator genes such as DNMT3A, TET2, and ASXL1 in hematopoietic stem cells (HSCs) and is associated with the risk of developing hematological malignancies such as myelodysplastic syndrome (MDS). Dysregulated immune and inflammatory signaling pathways have been implicated in hematopoietic abnormalities related to aging, cardiovascular disease, leading to MDS development. IL-6 signaling dependent pre-leukemic myeloproliferation was shown in Tet2-deficient mice upon DSS challenge, and a recent epidemiological sequencing study showed that inflammatory environment of ulcerative colitis promoted the positive selection of hematopoietic clones with DNMT3A mutation. However, it needs to be clarified how chronic UC contributes to hematopoiesis. So, it was investigated whether chronic UC drives clonal hematopoiesis in pre-leukemic Dnmt3a mutant mice.8-10 weeks old Dnmt3afl / flMx1Cre+mice were injected with poly I:C to generate Dnmt3a mutant (D3a- / -) mice. C57BL / 6 (B6) mice were also treated with poly I:C and used as control mice. After 2 weeks of poly I:C, D3a- / -and B6 mice were treated with and without 3% DSS for 3 cycles of 5 days with DSS and 7 days of resting between each cycle. It is worth noting here that D3a- / -mice were more frequently dying with 3% DSS; therefore, 2% DSS was used in all experiments with D3a- / -mice. After the 3rd cycle of DSS, mice were sacrificed and analyzed for the role of chronic UC in hematopoiesis. D3a- / -mice with chronic UC showed increased BM cellularity as compared to B6 mice with chronic UC (FIG.15A). Although, the frequency of LSKs was comparable between D3a- / -and B6 mice with chronic UC (FIGS.15B & 15C); however, the absolute number of LSKs was significantly increased in the BM of D3a- / -chronic UC mice (FIG.16A). Furthermore, within LSK cell fractions, the frequencies as well as the absolute cell numbers of LT-HSCs and MPPs (FIGS.15D & 15E) were significantly reduced, while the frequency and the absolute number of myeloid-biased HPC-2 cells (FIG.15F) were significantly enriched in the BM of chronic UC D3a- / -mice compared to chronic UC B6 mice. Consistent with enriched myeloid-biased HPC-2 cells in the BM, analysis of HSPCs showed enrichment in the frequencies and the absolute numbers of LKs and GMPs in the BM (FIG.15G & 15H and FIGS.16E & 16F) and the frequencies of LKs and GMPs (FIGS.16G & 16H) in the spleen; whereas, the frequency of MEPs was decreased, both in the BM and spleen of chronic UC D3a- / -mice compared to chronic UC B6 mice (FIG.15I and FIG.16I). To interrogate if mice bearing D3a- / -mutant clones exhibited exacerbated pro-inflammatory cells upon onset of chronic UC, the immune cells in the BM, spleen, and colon of mice treated with and without DSS were analyzed. The frequency of Ly6Chi monocytes was significantly reduced and the frequency of Ly6Clo monocytes was increased in the BM of chronic UC mice bearing D3a- / -mutant clones compared to control B6 mice (FIG.15J); whereas, the frequency of proinflammatory Ly6Chi monocytes was remarkably elevated and the frequency of anti-inflammatory Ly6Clo monocytes was reduced in the spleens of chronic UC mice bearing D3a- / -mutant clones compared to control chronic UC B6 mice (FIG.15K). Additionally, chronic UC mice bearing D3a- / -mutant clones displayed an enrichment in the frequency of CD11b+Gr-1+neutrophils (FIGS.16J & 16K) and an enormous reduction in the frequency of B220+B cells (FIGS.16L & 16M) both, in the BM and spleen compared to control chronic UC B6 mice. Furthermore, chronic UC mice bearing D3a- / -mutant clones showed an elevated accumulation of CD11b+Gr-1+neutrophils and CD3+T cells in the colon compared to control chronic UC B6 mice (FIGS.16N & 16O). These data indicate that D3a- / -mice had accumulated more proinflammatory immune cells during chronic ulcerative colitis, which is suggestive of accelerated colitis in D3a- / -mice. Having shown that HSCs derived from chronic UC B6 mice were functionally impaired under the transplant setting (FIGS.9C-9E), it was intended to understand what the functional fate of HSCs bearing D3a- / -mutation could be in a condition like chronic UC under the transplant setting. Engraftment of donor cells was determined in the PB at an interval of 4 weeks by flow cytometry analysis. The engraftment ability of stem cells bearing D3a- / -clones was reduced more profoundly in the PB, BM, and spleen compared to non-mutant stem cells from B6 control mice under the settings of chronic UC (FIGS.15L-15P). At the 24th week post transplantation, mice were analyzed for proportions of HSCs / HSPCs, including mature cells in the BM and spleen. It was found that within CD45.2+fraction, the frequencies of LSKs and myeloid- biased HPC-2 cells that are downstream of MPPs (FIGS.17A & 17D), were increased; whereas, the frequencies of LT-HSCs and MPPs (FIGS.17B & 17C) were reduced in the BM of recipient mice transplanted with CD45.2+stem cells from chronic UC mice bearing D3a- / -clones as compared to control chronic UC B6 mice. The frequency of MEPs was reduced in the BM of recipient mice transplanted with stem cells from chronic UC mice bearing D3a- / -clones compared to the control chronic UC B6 recipient mice (FIG.17E). This data indicates that under chronic UC conditions, only few a D3a- / -HSCs engraft that give rise to more myeloid progenitors, and less lymphoid and erythroid progenitor cells in the transplant setting. From these results, it could be inferred that chronic UC contributes to the manifestation of age-associated hematopoietic stem cell phenotypes in D3a- / -mice. At the mature cell level, it was found that CD11b+Gr-1+neutrophils were accumulated, and B220+B cells were reduced in the BM of recipient mice who had received CD45.2+stem cells from chronic UC mice bearing D3a- / -clones compared to the control chronic UC B6 recipient mice (FIGS.17F & 17G). Also in transplant setting, immature CD11b+c-Kit+myeloid blast cells were found to be accumulated in the BM and spleen of recipient mice who had received CD45.2+stem cells from chronic UC mice bearing D3a- / -clones compared to the control chronic UC B6 recipient mice (FIGS.17H & 17I). Collectively, the data indicate that chronic UC- induced aberrant hematopoiesis led to the differentiation of HSCs toward expansion of myelopoiesis and suppression of lymphopoiesis and erythropoiesis in pre-leukemic Dnmt3a mutant mice. Additionally, these data also highlighted the fact that chronic UC impairs engraftment potential of HSCs bearing D3a- / -clones and contributes to the manifestation of age-associated stem cell phenotypes. APE1 / Ref-1 inhibitor, APX3330, restores chronic UC driven defective hematopoiesis and mitigates gut damage in B6 mice and D3a mutant mice in an APE1 / Ref-1 / HIF-1α axis dependent manner. It has now been demonstrated that the blockade of the redox-activity of APE1 / Ref-1 with APX3330 restores defective hematopoiesis and mitigates chronic UC induced gut damage in B6 mice. Therefore, whether the ref-1 inhibitor, APX3330, would also effectively reinstate chronic UC associated phenotypes in mice bearing D3a mutation was analyzed. The treatment of B6 and D3a- / -mice was performed as shown in the FIG.18A. First, it was observed that the colon size was comparable between B6 and D3a- / -chronic UC mice (FIGS.18B & 18C); however, the colon weight, and colon weight / length ratio were increased in D3a- / -chronic UC mice (FIGS.18D & 18E), indicating that compared to B6 control mice, D3a- / -mice develop more pronounced inflammation in their gut during the chronic UC. As expected, ref-1 inhibitor, APX3330 treatment effectively restored the colons in both B6 and D3a- / -chronic UC mice (FIGs. 18B-18E). Then we examined the effect of chronic UC in the BM and found that the BM cellularity in chronic UC mice bearing D3a- / -clones was elevated, while this increase in the BM cellularity was reversed by the APX3330 treatment (FIG.19A). Along with this result, the APX3330 treatment led to normalizing the frequency of LSK-HSCs in the BM of chronic UC D3a- / - and B6 mice (FIG.19B). In UC patients and experimental mouse model, intestinal bleeding is one of the recurrent pathological conditions that may result in blood loss leading to anemia. Since MEPs produce red blood cells (RBCs) and platelets, c-Kit+HSPCs were interrogated for MEPs. Similar to the previous data (FIG. 15I), chronic UC D3a- / -mice showed reduced frequency of MEPs in the BM compared to chronic UC B6 mice. Furthermore, there was a decrease in the frequency of MEPs in D3a- / -mice under chronic UC reflected in the peripheral blood counts as D3a- / -mice displayed robust reduction in RBCs, hemoglobin (HGB), hematocrit (HCT), and an increase in the red cell distribution width-coefficient variation % (RDW-CV%) (FIGS. 19D-19G), which are indicative of infection related anemia in D3a- / -mice than in control B6 mice under chronic UC conditions. Interestingly, the frequency of MEPs and anemia associated with chronic UC were corrected in D3a- / -and B6 mice by the administration of APX3330 to levels observed in the veh group (FIGS.19C-19G). To examine if the APX3330 administration also corrected the abnormal proportions of immune cells, the PB, BM and spleens from mice in this Example were analyzed. First, the blood counts were analyzed and found that the absolute number and frequency of neutrophils and monocytes were elevated, while lymphocytes were reduced in the PB of chronic UC D3a- / -mice than in control chronic UC B6 mice (FIGs.19H- 19K). Furthermore, consistent with the PB counts, our flow cytometry analysis showed that CD11b+Gr-1+neutrophils were markedly accumulated (FIGS.18F-18H); whereas, B220+B cells were reduced (FIGS.18I-18K) in the PB, BM, and SP of D3a- / -mice than in B6 mice under chronic UC settings. Consistently, the blockade of redox-activity of ref- 1 with APX3330 restored the chronic UC-induced abnormal myeloid and lymphocytes in the BP, BM and spleen of chronic UC D3a- / -and B6 control mice (FIGS.19H-19K and FIGS.18F-18K). In a subsequent analysis, the APX3330 administration partially reduced the chronic UC driven splenomegaly (FIGS.19L & 19N), and immature myeloid-blast (CD11b+c-Kit+) cells (FIG.19O) in both D3a- / -and B6 mice. By contrast, PB and the BM of chronic ulcerative colitic D3a- / -and B6 mice did not reveal any changes in the accumulation of CD11b+c-Kit+myeloid-blast cells (FIGS.18L & 18M). To interrogate if chronic UC had also contributed to the expression of HIF-1α in CD34+HSPCs from the BM of D3a- / -mice as it was observed in chronic UC B6 mice (Fig.6B), and the administration of APX3330 would correct the expression of HIF-1α in HSPCs from the BM of D3a- / -mice. Intracellular staining and flow-cytometry analysis was utilized and revealed that the expression of HIF-1α in CD34+HSPCs from the BM of chronic UC D3a- / -mice was approximately 2-fold higher compared to chronic UC B6 mice; whereas, the blockade of the redox-activity of the ref-1 with APX3330, reduced the expression of HIF-1α in CD34+HSPCs to levels observed in veh-treated groups (FIG.19P). Altogether, these data suggest that APX3330 corrects chronic UC driven hematopoietic defects in D3a mutant mice and further highlight the facts that APE1 / Ref-1 / HIF-1α signaling cascade could serve as a better therapeutic target in the ulcerative colitis in the mutant settings of Dnmt3a.

Claims

CLAIMS What is claimed is:

1. A method of treating ulcerative colitis (UC) in a subject in need thereof, the method comprising administering to the subject an effective amount of an apurinic / apyrimidinic endonuclease 1 redox factor 1 (APE1 / Ref-1) inhibitor, pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof.

2. The method as set forth in claim 1, wherein the APE1 / Ref-1 inhibitor has the formula: Formula (I)wherein R1is selected from the group consisting of alkyl, alkoxy, hydroxyl, and hydrogen; R2 is an alkyl; R3and R6are independently selected from the group consisting of a substituted or unsubstituted alkoxy, a substituted or unsubstituted aryl and an oxo; R4 and R5 are independently selected from the group consisting of an alkoxy and aryl, or both R4 and R5taken together form a substituted or unsubstituted napthoquinone; X is selected from the group consisting of CH=CR2 and NCH, wherein R2 is selected from the group consisting of C1-C10 alkyl and CF3CH2CH2; and Y is selected from the group consisting of N(Rz)R2or NR^OR^, wherein each Rz is independently selected from the group consisting of C1-C6alkyl, heteroalkyl, cycloalkyl and cycloheteroalkyl, straight or branched chain or optionally substituted, or both Rz andR2taken together with the attached nitrogen form an optionally substituted heterocycle; where each R^ is independently selected from the group consisting of hydrogen, alkyl, heteroalkyl, cyclohexyl, and cycloheteroalkyl, each of which is optionally substituted, or both R^ are taken together with the attached nitrogen and oxygen to form an optionally substituted heterocycle.

3. The method as set forth in claim 1, wherein the APE1 / Ref-1 inhibitor is selected from an inhibitor set forth in Table 1.

4. The method as set forth in claim 1, wherein the APE1 / Ref-1 inhibitor is selected from the group consisting of 3-[(5-(2,3-dimethoxy-6-methy11,4-benzoquinoy1)]- 2-nony1-2-proprionic acid (APX3330), (2E)-2-[(3-methoxy-1,4-dioxo-1,4- dihydronapthalen -2-yl)methylidene]-N,N-dimethylpentanamide] (APX2007), [(2E)-2- [(3-methoxy-1,4-dioxo-1,4-dihydronaphthalen-2-yl)methylidene]-N,N- diethylpentanamide] (APX2009), (2E)-2-[(3-methoxy-1,4-dioxo-1,4-dihydronapthalen -2- yl)methylidene]-N-methoxypentanamide] (APX2014), (2E)-2-(3-methoxy-1,4-dioxo-1,4- dihydronaphthalen-2-yl)-N,N,2-trimethylprop-2-enamide (APX2032), pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof, and combinations thereof.

5. The method as set forth in claim 1, wherein the APE1 / Ref-1 inhibitor is APX3330 and the subject is administered from about 1.0 µM to about 50 µM APX3330.

6. The method as set forth in claim 1, wherein the APE1 / Ref-1 inhibitor is APX3330 and the subject is a human patient administered from about 10 mg / day to about 600 mg / day.

7. The method as set forth in claim 1 further comprising administering at least one additional therapeutic agent to the subject.

8. The method as set forth in claim 7, wherein the additional therapeutic agent is selected from the group consisting of 5-aminosalicylic acid (5-ASA), corticosteroids,azathioprine, 6-mercaptopurine, methotrexate, cyclosporine, tacrolimus, anti-TNF drugs vedolizumab, natalizumab, ustekinumab, probiotics, antibiotics, anti-inflammatories, and combinations thereof.

9. A method of treating inflammatory bowel disease (IBD) in a subject in need thereof, the method comprising administering to the subject an effective amount of an apurinic / apyrimidinic endonuclease 1 redox factor 1 (APE1 / Ref-1) inhibitor, pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof.

10. The method as set forth in claim 1, wherein the APE1 / Ref-1 inhibitor has the formula: Formula (I)wherein R1 is selected from the group consisting of alkyl, alkoxy, hydroxyl, and hydrogen; R2 is an alkyl; R3and R6are independently selected from the group consisting of a substituted or unsubstituted alkoxy, a substituted or unsubstituted aryl and an oxo; R4 and R5 are independently selected from the group consisting of an alkoxy and aryl, or both R4 and R5taken together form a substituted or unsubstituted napthoquinone; X is selected from the group consisting of CH=CR2 and NCH, wherein R2 is selected from the group consisting of C1-C10 alkyl and CF3CH2CH2; and Y is selected from the group consisting of N(Rz)R2 or NR^OR^, wherein each Rz is independently selected from the group consisting of C1-C6alkyl, heteroalkyl, cycloalkyl and cycloheteroalkyl, straight or branched chain or optionally substituted, or both Rz andR2taken together with the attached nitrogen form an optionally substituted heterocycle; where each R^ is independently selected from the group consisting of hydrogen, alkyl, heteroalkyl, cyclohexyl, and cycloheteroalkyl, each of which is optionally substituted, or both R^ are taken together with the attached nitrogen and oxygen to form an optionally substituted heterocycle.

11. The method as set forth in claim 9, wherein the APE1 / Ref-1 inhibitor is selected from an inhibitor set forth in Table 1.

12. The method as set forth in claim 9, wherein the APE1 / Ref-1 inhibitor is selected from the group consisting of 3-[(5-(2,3-dimethoxy-6-methy11,4-benzoquinoy1)]- 2-nony1-2-proprionic acid (APX3330), (2E)-2-[(3-methoxy-1,4-dioxo-1,4- dihydronapthalen -2-yl)methylidene]-N,N-dimethylpentanamide] (APX2007), [(2E)-2- [(3-methoxy-1,4-dioxo-1,4-dihydronaphthalen-2-yl)methylidene]-N,N- diethylpentanamide] (APX2009), (2E)-2-[(3-methoxy-1,4-dioxo-1,4-dihydronapthalen -2- yl)methylidene]-N-methoxypentanamide] (APX2014), (2E)-2-(3-methoxy-1,4-dioxo-1,4- dihydronaphthalen-2-yl)-N,N,2-trimethylprop-2-enamide (APX2032), pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof, and combinations thereof.

13. The method as set forth in claim 9, wherein the APE1 / Ref-1 inhibitor is APX3330 and the subject is administered from about 1.0 µM to about 50 µM APX3330.

14. The method as set forth in claim 9, wherein the APE1 / Ref-1 inhibitor is APX3330 and the subject is a human patient administered from about 10 mg / day to about 600 mg / day.

15. The method as set forth in claim 9 further comprising administering at least one additional therapeutic agent to the subject.

16. The method as set forth in claim 15, wherein the additional therapeutic agent is selected from the group consisting of 5-aminosalicylic acid (5-ASA), corticosteroids,azathioprine, 6-mercaptopurine, methotrexate, cyclosporine, tacrolimus, anti-TNF drugs vedolizumab, natalizumab, ustekinumab, probiotics, antibiotics, anti-inflammatories, and combinations thereof.

17. The method as set forth in claim 9, wherein the subject was administered corticosteroid.

18. The method as set forth in claim 9, wherein the subject is less than or equal to 45 years of age and has a high variant allele fraction (VAF) for myeloid-associated genes (M-CHIP) mutations.

19. The method as set forth in claim 9, wherein the subject is greater than 45 years of age and has an increased prevalence of M-CHIP mutations as compared to a healthy subject.

20. Use of an apurinic / apyrimidinic endonuclease 1 redox factor 1 (APE1 / Ref- 1) inhibitor, pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof in a medicament for treating ulcerative colitis (UC) or inflammatory bowel disease (IBD) in a subject in need thereof.

21. The use as set forth in claim 20, wherein the APE1 / Ref-1 inhibitor has the formula: Formula (I)wherein R1is selected from the group consisting of alkyl, alkoxy, hydroxyl, and hydrogen; R2 is an alkyl; R3 and R6 are independently selected from the group consisting of a substituted or unsubstituted alkoxy, a substituted or unsubstituted aryl and an oxo; R4 and R5are independently selected from the group consisting of an alkoxy and aryl, or both R4and R5taken together form a substituted or unsubstituted napthoquinone; X is selected from the group consisting of CH=CR2 and NCH, wherein R2 is selected from the group consisting of C1-C10alkyl and CF3CH2CH2; and Y is selected from the group consisting of N(Rz)R2or NR^OR^, wherein each Rz is independently selected from the group consisting of C1-C6 alkyl, heteroalkyl, cycloalkyl and cycloheteroalkyl, straight or branched chain or optionally substituted, or both Rz and R2taken together with the attached nitrogen form an optionally substituted heterocycle; where each R^ is independently selected from the group consisting of hydrogen, alkyl, heteroalkyl, cyclohexyl, and cycloheteroalkyl, each of which is optionally substituted, or both R^ are taken together with the attached nitrogen and oxygen to form an optionally substituted heterocycle.

22. The use as set forth in claim 20, wherein the APE1 / Ref-1 inhibitor is selected from an inhibitor set forth in Table 1.

23. The use as set forth in claim 20, wherein the APE1 / Ref-1 inhibitor is selected from the group consisting of 3-[(5-(2,3-dimethoxy-6-methy11,4-benzoquinoy1)]- 2-nony1-2-proprionic acid (APX3330), (2E)-2-[(3-methoxy-1,4-dioxo-1,4- dihydronapthalen -2-yl)methylidene]-N,N-dimethylpentanamide] (APX2007), [(2E)-2- [(3-methoxy-1,4-dioxo-1,4-dihydronaphthalen-2-yl)methylidene]-N,N- diethylpentanamide] (APX2009), (2E)-2-[(3-methoxy-1,4-dioxo-1,4-dihydronapthalen -2- yl)methylidene]-N-methoxypentanamide] (APX2014), (2E)-2-(3-methoxy-1,4-dioxo-1,4- dihydronaphthalen-2-yl)-N,N,2-trimethylprop-2-enamide (APX2032), pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof, and combinations thereof.

24. The use as set forth in claim 20, wherein the APE1 / Ref-1 inhibitor is APX3330 and the subject is administered from about 1.0 µM to about 50 µM APX3330.

25. The use as set forth in claim 20, wherein the APE1 / Ref-1 inhibitor is APX3330 and the subject is a human patient administered from about 10 mg / day to about 600 mg / day.

26. The use as set forth in claim 20 further comprising use of at least one additional therapeutic agent in combination with the APE1 / Ref-1 inhibitor.

27. The use as set forth in claim 26, wherein the additional therapeutic agent is selected from the group consisting of 5-aminosalicylic acid (5-ASA), corticosteroids, azathioprine, 6-mercaptopurine, methotrexate, cyclosporine, tacrolimus, anti-TNF drugs vedolizumab, natalizumab, ustekinumab, probiotics, antibiotics, anti-inflammatories, and combinations thereof.

28. The use as set forth in claim 20, wherein the subject is less than or equal to 45 years of age and has a high variant allele fraction (VAF) for myeloid-associated genes (M-CHIP) mutations.

29. The use as set forth in claim 20, wherein the subject is greater than 45 years of age and has an increased prevalence of M-CHIP mutations as compared to a healthy subject.

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