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到几年才能解决，从而导致 明显的发病率。有助于治愈这些病变的药理学干预措施将减少发病率，但 调节利什曼原虫病变修复的机制尚不清楚。此外，还有 我们对先天免疫核苷酸结合结构域和富含亮氨酸的重复的作用几乎没有投资 受体（NLR）家庭成员控制伤口愈合。使用皮肤利什曼病的鼠标模型， 我们已经发现NLR家族成员NLRP10在受感染伤口的愈合中的新作用，因为缺乏小鼠 与野生型小鼠相比，NLRP10会产生更大的病变，无法愈合。虽然大多数NLR家族的研究 在该模型中，模式识别受体在造血细胞中鉴定了这些受体的作用 NLRP10调节非脊髓性细胞群体的伤口愈合。重要的是，何时病变 NLPRP10缺陷型小鼠大于野生型小鼠的小鼠，这并不是由于无法控制 寄生虫，因为两组之间的寄生虫燃烧没有差异。相反，无菌伤口的愈合 在NLRP10缺陷小鼠中是完整的，这表明对NLRP10的要求是特定于存在的 伤口中的致病微生物。在此提案中，我们旨在确定NLRP10的机制 调节皮肤利什曼病的伤口修复。我们发现NLRP10缺乏成纤维细胞具有 钝增殖。在评估失败的原因时，我们确定这些成纤维细胞逮捕了他们 细胞周期的G0/G1阶段的增殖。我们还发现显示了显示的NLRP10缺乏成纤维细胞 β-半乳糖苷酶染色升高并增加了p16和p21 表达。 NLRP10缺乏性成纤维细胞也显示了感应的形态特征，并增加了 促炎性细胞因子和趋化因子的分泌，与这些具有感应的细胞一致 相关的秘密表型（SASP）。我们建议NLRP10在皮肤的成纤维细胞内作用 利什曼病病变可缓解感受并这样做有助于伤口修复反应。利用 P16-3MR鼠标模型，这将使​​我们能够识别并有选择地杀死Senscent细胞，我们将能够 定义皮肤利什曼病中感应对伤口修复的贡献，并确定NLRP10的贡献 调节此过程。我们将确定在l.乳杆菌中驱动成纤维细胞感应的因子 我们将进一步定义NLRP10对SASP的影响并确定因子 修复皮肤利什曼病伤所必需的。我们的研究将定义NLRP10在确定 衰老途径并确定新型的治疗靶标，以加速各种皮肤伤口愈合 病理。