Regulation of wound healing pathways by NLRP10 in cutaneous Leishmaniasis

NLRP10 对皮肤利什曼病伤口愈合途径的调节

• 批准号: 10783649
• 负责人: Fayyaz S. Sutterwala
• 金额: $ 40.41万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-18 至 2025-09-17
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10783649
• 关键词: Acceleration; Binding; Bone Marrow; CXCL1 gene; Cell Aging; Cell Cycle; Cell Cycle Arrest; Cells; Chimera organism; Chronic; Cutaneous; Cutaneous Leishmaniasis; Data; Disease; Endothelial Cells; Failure; Family; Family member; Fibroblasts; G1 Phase; Growth Factor; Hematopoietic; Immune; Immune response; Immunologics; Impaired healing; Impaired wound healing; Infection; Inflammatory; Interleukin-6; Intervention; Investigation; Leishmania; Leishmania major; Lesion; Leucine-Rich Repeat; Mediating; Modeling; Morbidity - disease rate; Morphology; Mus; Nodule; Nucleotides; Parasite Control; Parasites; Pathogenicity; Pathology; Pathway interactions; Patients; Pattern recognition receptor; Peptide Hydrolases; Phenotype; Play; Population; Process; Proliferating; Proteins; Protozoa; Reactive Oxygen Species; Regulation; Reporting; Resolution; Role; Signal Transduction; Skin; Skin repair; Skin wound healing; Stains; Sterility; Surface; Ulcer; Varicose Ulcer; Wild Type Mouse; Wound Infection; adaptive immune response; beta-Galactosidase; biological adaptation to stress; cell type; chemokine; chronic ulcer; chronic wound; conditional knockout; cytokine; defined contribution; diabetic ulcer; healing; high risk; in vivo; microorganism; mouse model; neutrophil; new therapeutic target; novel; pharmacologic; prevent; receptor; repaired; response; senescence; skin lesion; skin ulcer; skin wound; superinfection; tissue repair; wound; wound environment; wound healing

PROJECT SUMMARY Cutaneous leishmaniasis results in skin ulcerations that can take months to years to resolve resulting in significant morbidity. Pharmacologic interventions to aid in healing of these lesions would lessen morbidity, but the mechanisms that regulate the repair of Leishmania lesions are not well understood. In addition, there has been little investigation into the role of innate immune Nucleotide-Binding Domain and Leucine Rich Repeat Receptor (NLR) family members in regulating wound healing. Using a mouse model of cutaneous leishmaniasis, we have found a novel role for the NLR family member NLRP10 in healing of infected wounds, as mice that lack NLRP10 develop larger lesions that fail to heal compared to wild-type mice. While most studies of the NLR family of pattern recognitions receptors have identified roles for these receptors in hematopoietic cells, in this model NLRP10 regulates wound healing from a non-hematopoietic cell population. Importantly, while the lesions in NLPRP10-deficient mice were larger than those of wild-type mice, this was not due to a failure to control the parasite, as there was no difference in parasite burden between the groups. In contrast, healing of sterile wounds was intact in NLRP10-deficient mice, suggesting the requirement for NLRP10 is specific to the presence of pathogenic microorganisms in the wound. In this proposal, we aim to identify the mechanism by which NLRP10 regulates wound repair in cutaneous leishmaniasis. We have found that NLRP10-deficient fibroblasts have blunted proliferation. While evaluating the reason for this failure, we identified that these fibroblasts arrested their proliferation at the G0/G1 phase of the cell cycle. We also found NLRP10-deficient fibroblasts displayed increased markers of senescence, with elevated β-galactosidase staining and increased p16 and p21 expression. NLRP10-deficient fibroblasts also showed morphologic features of senescence and had increased secretion of pro-inflammatory cytokines and chemokines, consistent with these cells having a senescence- associated secretory phenotype (SASP). We propose that NLRP10 acts within fibroblasts in cutaneous leishmaniasis lesions to mitigate senescence and in doing so aids in the wound repair response. Utilizing the p16-3MR mouse model, which will allow us to both identify and selectively kill senescent cells, we will be able to define the contribution of senescence to wound repair in cutaneous leishmaniasis and determine how NLRP10 regulates this process. We will determine the factors in L. major lesions that drive fibroblast senescence in the absence of NLRP10. We will further define the influence of NLRP10 on the SASP and determine the factor required for repair of cutaneous leishmaniasis wounds. Our studies will define the role of NLRP10 in regulating senescence pathways and identify novel therapeutic targets to accelerate cutaneous wound healing in a variety of pathologies.

项目摘要 皮肤利什曼病导致皮肤溃疡，可能需要数月至数年才能解决， 严重的发病率。药物干预，以帮助愈合这些病变将减少发病率，但 调节利什曼原虫病损修复的机制还不清楚。此外， 关于天然免疫中核苷酸结合域和富含亮氨酸重复序列的作用， 受体（NLR）家族成员在调节伤口愈合中的作用。使用皮肤利什曼病的小鼠模型， 我们已经发现NLR家族成员NLRP 10在感染伤口愈合中的新作用，因为缺乏NLRP 10的小鼠， 与野生型小鼠相比，NLRP 10发展出更大的无法愈合的病变。虽然大多数关于NLR家族的研究 模式识别受体已经确定了这些受体在造血细胞中的作用，在这个模型中， NLRP 10调节来自非造血细胞群体的伤口愈合。重要的是，虽然病变在 NLPRP 10缺陷型小鼠比野生型小鼠大，这不是由于未能控制NLPRP 10缺陷型小鼠。 寄生虫，因为各组之间的寄生虫负担没有差异。相反，无菌伤口的愈合 在NLRP 10缺陷小鼠中是完整的，这表明对NLRP 10的需求是特异性的， 伤口中的病原微生物在这项建议中，我们的目的是确定NLRP 10 调节皮肤利什曼病的伤口修复。我们已经发现NLRP 10缺陷的成纤维细胞 钝增殖。在评估这种失败的原因时，我们发现这些成纤维细胞抑制了它们的生长。 在细胞周期的G 0/G1期增殖。我们还发现NLRP 10缺陷的成纤维细胞显示 衰老标志物增加，β-半乳糖苷酶染色升高，p16和p21增加 表情NLRP 10缺陷的成纤维细胞也显示出衰老的形态学特征， 促炎细胞因子和趋化因子的分泌，与这些具有衰老的细胞一致， 相关分泌表型（SASP）。我们认为NLRP 10在皮肤成纤维细胞中起作用， 利什曼病损害，以减轻衰老，这样做有助于伤口修复反应。利用 p16- 3 MR小鼠模型，这将使我们能够识别和选择性杀死衰老细胞，我们将能够 确定皮肤利什曼病中衰老对伤口修复的贡献，并确定NLRP 10 规范这个过程。我们将确定L中的因子。主要病变，驱动成纤维细胞衰老， 没有NLRP 10。我们将进一步确定NLRP 10对SASP的影响，并确定因子 用于皮肤利什曼病伤口的修复。我们的研究将确定NLRP 10在调节 衰老途径，并确定新的治疗靶点，以加速各种皮肤伤口愈合 的病理学。