Novel imaging approach to study podocyte function in vivo

研究体内足细胞功能的新成像方法

• 批准号: 9298641
• 负责人: JANOS PETI-PETERDI
• 金额: $ 37.13万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-15 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9298641
• 关键词: Actins; Acute; Address; Albumins; Albuminuria; Amplifiers; Angiotensin II; Animal Model; Applications Grants; Area; Binding; Blood Flow Velocity; Blood capillaries; Blood flow; Calcium; Caliber; Cells; Chronic Kidney Failure; Clinical; Comorbidity; Cytoskeleton; Development; Diagnostic; Disease; Environment; FDA approved; Focal Segmental Glomerulosclerosis; Foot Process; Functional disorder; Genetic; Health; Histology; Homeostasis; Human; Image; Immunoglobulin G; In Vitro; Injury; Kidney; Kidney Diseases; Knock-out; Label; Maintenance; Mediating; Microscopy; Modeling; Morphology; Mus; Organ; P2Y2 receptor; Pathogenesis; Pathology; Permeability; Pharmacology; Phenotype; Physiological; Purinergic P2 Receptors; Purinoceptor; Role; Sclerosis; Signal Transduction; Suramin; Testing; Therapeutic; Tissues; Treatment Efficacy; Tubular formation; capillary; cytotoxic; extracellular; glomerular filtration; glomerular function; glomerulosclerosis; hemodynamics; imaging approach; in vivo; innovation; microscopic imaging; mortality; mouse model; new therapeutic target; novel; novel therapeutic intervention; paracrine; podocyte; prevent; public health relevance; pyridoxal phosphate-6-azophenyl-2' ,4' -disulfonic acid; renal tubular transport; response; slit diaphragm

DESCRIPTION (provided by applicant): Glomerular dysfunction is a common basis for the development of chronic kidney disease (CKD), a condition with significant comorbidities and mortalities. Recent studies highlighted the role of podocyte actin cytoskeleton and the slit diaphragm in the maintenance of the glomerular filtration barrier (GFB), and the development of albuminuria (AU) and focal segmental glomerulosclerosis (FSGS). The key role of cytosolic calcium ([Ca2+]i) signaling in podocyte function and the above pathologies is established, however our mechanistic understanding of podocyte [Ca2+]i dynamics is limited to a few players (angiotensin II, TRPC5/6). Most P2 purinoceptors including P2Y2 that bind extracellular ATP signal via [Ca2+]i and have been implicated in a variety of (patho)physiological functions in many organs including paracrine cell-to-cell crosstalk, tissue injury and sclerosis. While the important function of the P2Y2 receptor in renal tubular transport and blood flow autoregulation is well established, its role in podocytes/glomerulus is largely unknown. Here we propose to study podocyte [Ca2+]i dynamics in vivo in the intact kidney in health and disease and to characterize the role and therapeutic relevance of a novel [Ca2+]i signaling mechanism in podocytes that is mediated by P2Y2 purinergic receptors. We hypothesize that P2Y2-mediated elevations in [Ca2+]i is a robust and key novel mechanism in primary podocyte injury, and also that its cell-to-cell propagation between podocytes results in amplified focal segmental dysfunction of the GFB and the development of AU and FSGS. The overall theme, to study the role of P2Y2 in podocyte/GFB injury and the effect of P2Y2 blockade on AU and FSGS, will be addressed by applying a novel imaging approach that employs intravital multiphoton microscopy (MPM) combined with new mouse models of fluorescent podocyte labeling and tagging. These studies may potentially change the current view of the pathogenesis of glomerular disorders and may also provide a clinically and immediately available, novel therapeutic approach for glomerular kidney diseases. The specific aims are to: (1) Characterize the role of P2Y2 signaling in podocyte [Ca2+]i dynamics and GFB/glomerular function in response to podocyte injury/disease. (2) Establish the amplifier function of P2Y2 signaling in the propagation of podocyte injury. (3) Test whether P2Y2 blockade can blunt/reduce the development of podocyte injury, AU and FSGS.

描述（由申请人提供）：肾小球功能障碍是慢性肾病（CKD）发展的常见基础，慢性肾病是一种具有显着合并症和死亡率的疾病。最近的研究强调了足细胞肌动蛋白细胞骨架和狭缝隔膜在维持肾小球滤过屏障（GFB）以及白蛋白尿（AU）和局灶节段性肾小球硬化（FSGS）的发展中的作用。胞浆钙 ([Ca2+]i) 信号传导在足细胞功能和上述病理学中的关键作用已经确定，但我们对足细胞 [Ca2+]i 动力学的机制理解仅限于少数参与者（血管紧张素 II、TRPC5/6）。大多数 P2 嘌呤受体（包括 P2Y2）通过 [Ca2+]i 结合细胞外 ATP 信号，并与许多器官的多种（病理）生理功能有关，包括旁分泌细胞间串扰、组织损伤和硬化。虽然 P2Y2 受体在肾小管转运和血流自动调节中的重要功能已得到充分证实，但其在足细胞/肾小球中的作用很大程度上未知。在这里，我们建议研究健康和疾病状态下完整肾脏中足细胞 [Ca2+]i 的体内动态，并表征足细胞中由 P2Y2 嘌呤能受体介导的新型 [Ca2+]i 信号传导机制的作用和治疗相关性。我们假设 P2Y2 介导的 [Ca2+]i 升高是原发性足细胞损伤中一种强有力且关键的新机制，并且其在足细胞之间的细胞间传播导致 GFB 局灶节段功能障碍放大和 AU 的发展和 FSGS。总体主题是研究 P2Y2 在足细胞/GFB 损伤中的作用以及 P2Y2 阻断对 AU 和 FSGS 的影响，将通过应用一种新颖的成像方法来解决，该方法采用活体多光子显微镜 (MPM) 结合新的荧光小鼠模型足细胞标记和标记。这些研究可能会改变目前对肾小球疾病发病机制的看法，也可能为肾小球肾脏疾病提供临床上立即可用的新治疗方法。具体目标是： (1) 表征 P2Y2 信号传导在足细胞 [Ca2+]i 动态和 GFB/肾小球功能中响应足细胞损伤/疾病的作用。 (2)建立P2Y2信号在足细胞损伤传播中的放大功能。 (3)测试P2Y2阻断是否可以减弱/减少足细胞损伤、AU和FSGS的发展。

项目成果

Novel fluorescence techniques to quantitate renal cell biology.

定量肾细胞生物学的新型荧光技术。

• 发表时间: 2019
• 作者: Shroff, Urvi Nikhil;Schiessl, Ina Maria;Gyarmati, Georgina;Riquier;Peti
• 通讯作者: Peti

Advances in Renal Cell Imaging.

肾细胞成像的进展。

• 发表时间: 2018
• 影响因子: 3.3
• 作者: Gyarmati, Georgina;Kadoya, Hiroyuki;Moon, Ju;Burford, James L;Ahmadi, Nariman;Gill, Inderbir S;Hong, Young;Dér, Bálint;Peti
• 通讯作者: Peti

Combined use of electron microscopy and intravital imaging captures morphological and functional features of podocyte detachment.

电子显微镜和活体成像的结合使用捕获足细胞脱离的形态和功能特征。

• 发表时间: 2017-08
• 作者: Burford, James L;Gyarmati, Georgina;Shirato, Isao;Kriz, Wilhelm;Lemley, Kevin V;Peti
• 通讯作者: Peti

Novel in vivo techniques to visualize kidney anatomy and function.

新颖的体内技术使肾脏解剖结构和功能可视化。

• 发表时间: 2015-07
• 影响因子: 19.6
• 作者: Peti;Kidokoro, Kengo;Riquier
• 通讯作者: Riquier

The role of TRPC6 calcium channels and P2 purinergic receptors in podocyte mechanical and metabolic sensing.

TRPC6 钙通道和 P2 嘌呤能受体在足细胞机械和代谢传感中的作用。

• 发表时间: 2021-12-16
• 影响因子: 1.4
• 作者: Gyarmati, Georgina;Toma, Ildikó;Izuhara, Audrey;Burford, James L;Shroff, Urvi Nikhil;Papadouri, Stella;Deepak, Sachin;Peti
• 通讯作者: Peti