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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要皮肤利什曼病会导致皮肤溃疡，可能需要数月至数年才能消退，从而导致帮助治愈这些病变的药物干预会降低发病率，但是此外，调节利什曼原虫病变修复的机制尚不清楚。对先天免疫核苷酸结合域和富含亮氨酸重复序列的作用的研究很少使用皮肤利什曼病小鼠模型调节伤口愈合的受体（NLR）家族成员，我们发现 NLR 家族成员 NLRP10 在感染伤口的愈合中具有新的作用，因为缺乏与野生型小鼠相比，NLRP10 产生更大的损伤，无法愈合，而大多数 NLR 家族的研究。模式识别受体已确定这些受体在造血细胞中的作用，在此模型中重要的是，NLRP10 调节非造血细胞群的伤口愈合，而病变则在其中。 NLPRP10缺陷型小鼠比野生型小鼠体型更大，这并不是由于未能控制寄生虫，因为各组之间的寄生虫负荷没有差异，相反，无菌伤口的愈合。在 NLRP10 缺陷小鼠中，NLRP10 是完整的，这表明对 NLRP10 的需求是特定于 NLRP10 的存在的。在本提案中，我们的目标是确定 NLRP10 的作用机制。我们发现 NLRP10 缺陷的成纤维细胞具有调节皮肤利什曼病伤口修复的作用。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增殖。我们还发现 NLRP10 缺陷的成纤维细胞显示出细胞周期 G0/G1 期的增殖。衰老标志物增加，β-半乳糖苷酶染色升高，p16 和 p21 增加 NLRP10 缺陷的成纤维细胞也表现出衰老的形态特征并增加。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我们提出 NLRP10 在皮肤成纤维细胞内发挥作用。利什曼病病变以减轻衰老，从而有助于伤口修复反应。 p16-3MR 小鼠模型，这将使我们能够识别并选择性杀死衰老细胞，我们将能够定义衰老对皮肤利什曼病伤口修复的贡献并确定 NLRP10 我们将确定 L.major 病变中驱动成纤维细胞衰老的因素。我们将进一步定义NLRP10对SASP的影响并确定该因素。我们的研究将确定 NLRP10 在调节中的作用。衰老途径并确定新的治疗靶点以加速各种皮肤伤口愈合病理学。