The Role of KLF15 as a transcriptional regulator of podocyte differentiation

KLF15 作为足细胞分化转录调节因子的作用

• 批准号: 8750137
• 负责人: Sandeep K Mallipattu
• 金额: $ 17.62万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8750137
• 关键词: AIDS-Associated Nephropathy; Activities of Daily Living; Address; Advisory Committees; American; Attenuated; Award; Binding Sites; Biochemistry; Biology; Biopsy Specimen; CREB1 gene; Characteristics; Chronic Kidney Failure; Data; Development; Development Plans; Dexamethasone; Diabetic Nephropathy; Differentiation Antigens; Disease; Educational Curriculum; Environment; Epithelial Cells; Etiology; Filtration; Focal Segmental Glomerulosclerosis; Foundations; Functional disorder; Funding; Genes; Glucocorticoid Receptor; Glucocorticoids; Goals; Grant; HIV-1; Hour; Human; In Vitro; Injury; Injury to Kidney; Interdisciplinary Study; Journals; Kidney; Kidney Diseases; Kidney Failure; Knock-out; Knockout Mice; Knowledge; Lymphocyte; Mediating; Mediator of activation protein; Mentors; Modeling; Molecular; Molecular Biology; Mus; NPHS2 protein; Nephrology; Nephrotic Syndrome; Outcomes Research; Pathology; Peer Review; Predisposition; Promoter Regions; Proteinuria; Publishing; Recovery; Regulation; Renal glomerular disease; Research; Research Personnel; Research Project Grants; Research Proposals; Response Elements; Role; Signal Pathway; Societies; System; Systems Biology; Techniques; Testing; Therapeutic; Therapeutic immunosuppression; Toxic effect; Training; Transgenic Mice; Tretinoin; Universities; Up-Regulation; Work; Writing; Yang; Zinc Fingers; career; career development; design; glomerular filtration; in vivo Model; insight; medical schools; meetings; multidisciplinary; nephrin; novel; overexpression; podocyte; promoter; protective effect; public health relevance; skills; stem; symposium; therapeutic target; transcription factor

DESCRIPTION (provided by applicant): The primary etiologies of Chronic Kidney Disease are a direct result of initial glomerular dysfunction. Podocytes are epithelial cells in the glomerulus that help maintain the renal filtration barrier. In many glomerular diseases, the podocyte loses specific markers of differentiation, characteristic morphologic features, and the functional capacity to maintain the glomerular filtration barrier. We recently characterized the role of Kr¿ppel-Like Factor 15 (KLF15), a kidney-enriched ubiquitous transcription factor, in podocyte differentiation. Specifically, we showed that a global loss of KLF15 increased the susceptibility to kidney injury in murine models of podocyte injury. Furthermore, we confirmed that the local kidney expression of KLF15 is reduced in human glomerular diseases such as FSGS and HIVAN. Finally, we also established that KLF15 is required for recovery from podocyte injury. Retinoic Acid (RA) has previously been shown to attenuate the loss of podocyte differentiation markers and thereby reduce proteinuria and ameliorate kidney disease. We have shown that RA-induced podocyte differentiation is mediated by KLF15. Glucocorticoids (GCs) are the first line of immunosuppressive therapy in the treatment of many glomerular diseases. Similar to Retinoic Acid, GCs have also been shown to protect podocytes from injury. Others have identified GC response elements in the promoter region of KLF15. We observed that dexamethasone increased KLF15 expression in human podocytes and in primary murine podocytes in culture. In addition, a loss of KLF15 in primary podocytes in culture attenuated the increase in GC-induced podocyte differentiation. Combined, we hypothesize that KLF15, a key transcriptional regulator of podocyte differentiation, mediates the renal protective effects of RA and GCs in glomerular disease. To address the hypothesis, we propose to first determine that KLF15 is necessary for podocyte differentiation using podocyte specific knockout mice and inducible, podocyte-specific KLF15 overexpression murine model. Second, we plan to ascertain the role of KLF15 in mediating renal protective effects of RA and GCs in glomerular disease. Finally, we propose to identify the upstream factors regulating KLF15 and downstream factors regulated by KLF15 in podocyte differentiation. Impact of proposed research outcome: We plan to determine whether the up-regulation of KLF15 mediates the renal protective effect of glucocorticoids and retinoic acid in glomerular disease. This will provide new insight into the podocyte biology and pathology, as well as a potential new target for therapy. Candidate's short-term and long-term goals: My short-term goal is to meet the specific aims of this research project while I continue to expand my knowledge on podocyte biology and disease by attending formal weekly divisional research conferences and monthly Work in Progress meetings. I will also attend the annual American Society of Nephrology national meeting and the biennial Podocyte meeting and FASEB KLF meeting, where I will be exposed to the latest research in the field of glomerular disease and KLF biology. I will continue to take opportunities such as these to share my data so that I can benefit in my professional development from the criticism of my colleagues in our field. Justification of the need for further training: My current research project involves the role of KLF15 in podocyte differentiation. With the K08 Award, I will be expected to expand my knowledge in molecular biology, biochemistry, and regulatory networks by attending graduate level courses at Stony Brook University School of Medicine. In addition, further mentoring from the multidisciplinary advisory committee during the next five years will be crucial in providing a strong foundation towards research independence. I am prepared to spend 75% of my professional effort towards reaching the goals outlined in my research strategy and training plan. My Long-term goal is to be an expert in KLF15 in kidney disease, specifically to gain a strong foundation in utilizing an integrative approach of molecular techniques and systems biology to identify potential therapeutic targets in glomerular disease. I can achieve this by completing the outlined training goals and the specific aims of my "research strategy." In addition, as I meet my research goals, I will work on publishing my work in peer-reviewed journals and apply for foundation grants with original ideas that may stem from my current work. After four years of support from the career-development grant, I plan to begin my career as an independent investigator by applying for R01 level funding. Candidate's career development plan and environment: I have designed a multidimensional training plan to prepare me for my interdisciplinary research proposal. In addition, my mentors (Dr. Yang and Dr. He) and I have assembled multi-disciplinary panel of experts to serve on an advisory committee (Dr. Lieberthal, Dr. D'Agati, Dr. Schl¿ndorff, and Dr. Ma'ayan). We have also devised a training curriculum encompassing 4 distinct modules: 1) Mastering molecular techniques to study the KLF15 signaling pathway, 2) Murine models of kidney injury and pathology, 3) Regulatory Networks in podocyte biology, and 4) Career development and grant writing skills.

描述（由申请人提供）：慢性肾病的主要病因是最初肾小球功能障碍的直接结果，足细胞是肾小球中的上皮细胞。 在许多肾小球疾病中，足细胞失去了特定的分化标记、特征性形态特征和维持肾小球滤过屏障的功能能力。 ppel 样因子 15 (KLF15) 是足细胞分化中富含肾脏的普遍转录因子。具体来说，我们发现 KLF15 的整体缺失会增加小鼠足细胞损伤模型对肾损伤的易感性。在人类肾小球疾病（例如 FSGS 和 HIVAN）中，KLF15 的局部肾脏表达降低。最后，我们还确定 KLF15 是足细胞恢复所必需的。先前已证明视黄酸 (RA) 可以减轻足细胞分化标记物的损失，从而减少蛋白尿并改善肾脏疾病。我们已经证明，糖皮质激素 (GC) 是 RA 诱导的足细胞分化的一线药物。与视黄酸类似，GC 也被证明可以保护足细胞免受免疫抑制治疗的影响。其他人已经鉴定出 KLF15 启动子区域的 GC 反应元件，我们观察到地塞米松增加了人足细胞和培养的原代小鼠足细胞中的 KLF15 表达。此外，培养物中原代足细胞中 KLF15 的缺失减弱了 GC 的增加。结合诱导足细胞分化，我们发现足细胞分化的关键转录调节因子 KLF15 介导 RA 和 GC 的肾脏保护作用。为了解决这一假设，我们建议首先使用足细胞特异性敲除小鼠和可诱导的足细胞特异性 KLF15 过表达小鼠模型确定 KLF15 对于足细胞分化是必需的。其次，我们计划确定 KLF15 在介导肾脏保护中的作用。最后，我们建议确定调节 KLF15 的上游因素和受 KLF15 调节的下游因素。拟议研究结果的影响：我们计划确定 KLF15 的上调是否介导糖皮质激素和视黄酸在肾小球疾病中的肾脏保护作用，这将为足细胞生物学和病理学以及足细胞分化提供新的见解。候选人的短期和长期目标：我的短期目标是实现该研究项目的具体目标，同时继续扩展我在足细胞生物学和足细胞生物学方面的知识。我还将参加一年一度的美国肾病学会全国会议和两年一次的足细胞会议和 FASEB KLF 会议，在那里我将接触到该领域的最新研究。我将继续利用此类机会分享我的数据，以便我能够从我们领域同事的批评中受益：我当前的研究项目。涉及到凭借 K08 奖，我将通过参加石溪大学医学院的研究生课程来扩展我在分子生物学、生物化学和调控网络方面的知识，此外，还将获得多学科的进一步指导。未来五年的咨询委员会对于为研究独立性提供坚实的基础至关重要，我准备花费 75% 的专业努力来实现我的研究策略和培训计划中概述的目标。成为专家在肾脏疾病的 KLF15 中，特别是为了利用分子技术和系统生物学的综合方法来确定肾小球疾病的潜在治疗靶点奠定坚实的基础，我可以实现这一目标。 通过完成概述的培训目标和我的“研究策略”的具体目标，此外，当我实现我的研究目标时，我将致力于在同行评审的期刊上发表我的作品，并以可能产生的原创想法申请基金会资助。经过四年的职业发展补助金的支持，我计划通过申请 R01 级别的资助来开始我的职业生涯：我设计了一个多维的培训计划来准备。我的跨学科研究此外，我和我的导师（杨博士和何博士）组建了一个多学科专家小组，担任顾问委员会成员（Dr. Lieberthal、D'Agati 博士、Schl¿ndorff 博士和Ma'ayan 博士）。我们还设计了包含 4 个不同模块的培训课程：1) 掌握研究 KLF15 信号通路的分子技术，2) 肾损伤和病理学的小鼠模型，3) 监管足细胞生物学网络，以及 4) 职业发展和资助写作技能。