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脏疾病（CKD）发展的常见基础，慢性肾脏疾病（CKD）的合并症和死亡率很高。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）以及蛋白尿（AU）和局灶性节段性段性肾小球乳化度（FSG）的发展中的作用。胞质钙（[Ca2+] i）信号在足细胞功能和上述病理中的关键作用是建立的，但是我们对足细胞[Ca2+] I动力学的机械理解仅限于一些玩家（血管紧张素II，TRPC5/6）。大多数P2 Purinoceptor在包括[CA2+] I的P2Y2中结合细胞外ATP信号的大多数P2Y2，并且在许多器官中都与多种（病原）生理功能有关，包括旁分泌细胞到细胞 - 细胞 - 细胞串扰，组织损伤和硬化症。虽然P2Y2受体在肾小管运输和血流自动调节中的重要功能已经很好地确定，但其在足细胞/肾小球中的作用在很大程度上尚不清楚。在这里，我们建议研究健康和疾病完整肾脏中的Podocyte [Ca2+] I动力学，并表征新颖的[Ca2+] I信号传导机制在P2Y2嘌呤能受体介导的Podocytes中的作用和治疗相关性。我们假设[Ca2+] I中的P2Y2介导的升高是原发性足细胞损伤中强大而关键的新型机制，并且其足细胞之间的细胞间传播会导致GFB的局灶性节段性分段和AU和FSG的发展。总体主题是研究P2Y2在Podocyte/GFB损伤中的作用以及P2Y2阻滞对AU和FSG的影响，将通过应用一种新型的成像方法来解决，该方法采用了新型的成像方法，该方法采用了插入式多光（MPM）与荧光podocyte和标记的新小鼠模型相结合。这些研究可能会改变肾小球疾病发病机理的当前观点，还可能为肾小球肾脏疾病提供临床和立即可用的新型治疗方法。具体目的是：（1）表征P2Y2信号在足细胞[Ca2+] I动力学和GFB/肾小球功能中的作用，以响应足细胞损伤/疾病。 （2）在足细胞损伤传播中建立P2Y2信号的放大器功能。 （3）测试P2Y2封锁是否可以钝化/减少足细胞损伤，AU和FSG的发展。

项目成果

An ectopic renin-secreting adrenal corticoadenoma in a child with malignant hypertension.

恶性高血压儿童的异位肾素分泌性肾上腺皮质腺瘤。

• DOI: 10.14814/phy2.12728
• 发表时间: 2016
• 期刊: Physiological reports
• 影响因子: 2.5
• 作者: Kaslow,AbrahamM;Riquier-Brison,Anne;Peti-Peterdi,Janos;Shillingford,Nick;HaDuong,Josephine;Venkatramani,Rajkumar;Gayer,ChristopherP
• 通讯作者: Gayer,ChristopherP

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

• DOI: 10.1007/s00424-017-2020-0
• 发表时间: 2017-08
• 期刊: Pflugers Archiv : European journal of physiology
• 作者: Burford JL;Gyarmati G;Shirato I;Kriz W;Lemley KV;Peti-Peterdi J
• 通讯作者: Peti-Peterdi J

In vivo microscopy.

活体显微镜检查。

• DOI: 10.1016/j.nephro.2016.01.004
• 发表时间: 2016
• 期刊: Nephrologie & therapeutique
• 影响因子: 0.7
• 作者: Peti-Peterdi,János

A practical new way to measure kidney fibrosis.

一种测量肾纤维化的实用新方法。

• DOI: 10.1016/j.kint.2016.07.036
• 发表时间: 2016
• 期刊: Kidney international
• 影响因子: 19.6
• 作者: Peti-Peterdi,János

On the Origin of Urinary Renin: A Translational Approach.

• DOI: 10.1161/hypertensionaha.115.07012
• 发表时间: 2016-05
• 期刊: Hypertension (Dallas, Tex. : 1979)
• 作者: Roksnoer LC;Heijnen BF;Nakano D;Peti-Peterdi J;Walsh SB;Garrelds IM;van Gool JM;Zietse R;Struijker-Boudier HA;Hoorn EJ;Danser AH
• 通讯作者: Danser AH