(PQ12) Targeting SMPDL3b to Prevent Radiation-Induced Nephrotoxicity

(PQ12) 靶向 SMPDL3b 预防辐射引起的肾毒性

• 批准号: 10442691
• 负责人: BRIAN MARPLES
• 金额: $ 37.41万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10442691
• 关键词: ANGPT1 gene; Acids; Actin-Binding Protein; Actins; Acute; Affect; Agonist; Apoptosis; Atrophic; Bilateral; Binding; Biological Markers; Blood; Blood Vessels; C57BL/6 Mouse; Cancer Patient; Cell Culture Techniques; Cell Survival; Cell membrane; Cells; Ceramides; Chronic; Cisplatin; Clinical; Clinical Treatment; Coculture Techniques; Combined Modality Therapy; Complication; Creatinine; Cytosol; Data; Development; Dose; Doxycycline; EDN1 gene; Endothelial Cells; Endothelin-1; Endothelium; Engineering; Enzymes; Fibrosis; Fractionated radiotherapy; Functional disorder; Goals; Homeostasis; Human; Impaired Renal Function; In Vitro; Inflammation; Injury; Injury to Kidney; Kidney; Kidney Diseases; Kidney Failure; Knock-out; Knockout Mice; Knowledge; Link; Lipids; MS4A1 gene; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Mediator of activation protein; Modality; Molecular; Molecular Target; Monoclonal Antibodies; Morbidity - disease rate; Morphology; Mus; Organ; Oxidative Stress; Pathway interactions; Placebo Control; Play; Proteinuria; Quality of life; Radiation; Radiation Injuries; Radiation Tolerance; Radiation induced damage; Radiation therapy; Radio; Regulation; Renal function; Retreatment; Role; Serum; Severities; Signal Transduction; Solid; Sphingolipids; Sphingomyelinase; Sphingosine; Testing; Time; Tissues; Tubular formation; Urine; Vascular Endothelial Growth Factors; Western Blotting; base; cancer radiation therapy; cell injury; cellular targeting; chemokine; chemotherapy; clinical development; clinically relevant; cytokine; ezrin; glomerular endothelium; image guided; improved; in vivo; irradiation; kidney cortex; kidney dysfunction; kidney vascular structure; mouse model; nephrotoxicity; novel; overexpression; paracrine; podocyte; prevent; protein expression; radiation mitigation; radiation mitigator; receptor; rituximab; sphingosine 1-phosphate; standard of care; targeted agent

PROJECT SUMMARY Radiation nephropathy (RN) is less common than chemotherapy-induced nephrotoxicity but still represents a serious late complication after radiation therapies for cancer. RN is irreversible and no effective clinical treatments exist to prevent RN or ameliorate radiation-associated kidney injury. Podocyte loss, tubular atrophy and endothelial damage have been linked with RN, but the molecular mechanisms governing RN are not known. We discovered that the enzyme sphingomyelin-phosphodiesterase-acid-like-3b (SMPDL3b) is an important regulator of radiation damage in renal podocytes after single dose (SD) radiotherapy (RT). Radiation damage reduced SMPDL3b expression triggering the cellular relocation of ezrin and a morphological change that altered podocyte functionality. Treatment with rituximab, which we demonstrated to bind SMPDL3b and to protect podocyte morphology, reduced SD RT induced RN in C57BL/6 mice but not in our newly-developed conditional podocyte-specific SMPDL3b knock-out mice. Based on these data we hypothesize that sphingolipids play a vital role in radiation-induced podocytopathy which governs RN. The objective is to investigate the mechanistic role of SMPDL3b in renal injury after fractionated low-dose radiotherapy (F-RT) with concurrent cisplatin (CDDP) as this represents a standard of care for many solid cancers. Our long-term goal is to discover a molecular-based protective or mitigating strategy for RN, and potentially chemotherapy-induced nephrotoxicity. We will test our hypothesis with the following three specific aims using a combined in vivo-in vitro approach: Aim 1: To determine if SMPDL3b regulates severity and latency of RT-associated kidney injury and functional RN after clinically-relevant F-RT, CDDP and concurrent F-RT+CDDP. This aim will also explore the role of SMPDL3b in tissue tolerance for RT retreatment injury, using C57BL/6 mice and our unique SMPDL3b-knockout and SMPDL3b-inducible mouse models. Aim 2: To determine the mechanism by which podocyte expression of SMPDL3b affects RT-mediated podocyte and glomerular endothelial cell (GEC) injury. We hypothesize that SMPDL3b affects RT induced compartmentalization of podocyte ezrin and affects GECs via altered endothelin-1 (EDN1) and END1 receptor type A (EDNRA) cross talk. GEC survival after RT will be studied by co-culturing GECs with podocytes lacking or expressing SMPDL3b. Aim 3: To determine if targeting sphingolipids prevents RN. We will investigate if protection of SMPDL3b or S1P will avert long-term functional renal injury in C57BL/6 mice after F-RT, CDDP and F-RT+CDDP. Mechanisms will be confirmed using our unique SMPDL3b-knockout and SMPDL3b-inducible mouse models. The findings from these studies will be significant because they offer the potential for molecular-targeted mitigation for RN, and radiation-associated kidney injury, after RT and combined modality injury.

项目摘要 辐射肾病（RN）不如化学疗法诱导的肾毒性常见，但仍代表 癌症放射治疗后严重的晚期并发症。 RN是不可逆的，没有有效的临床 存在治疗以防止RN或改善辐射相关的肾脏损伤。足细胞损失，管状萎缩 内皮损伤与RN有关，但尚不清楚控制RN的分子机制。 我们发现酶鞘磷脂 - 磷酸二酯酶 - 酸样-3B（SMPDL3B）是一个重要的 单剂量（SD）放疗（RT）后肾足细胞中辐射损伤的调节剂。辐射损坏 降低的SMPDL3B表达触发了Ezrin的细胞重定位和形态学变化，改变了 足细胞功能。用利妥昔单抗治疗，我们证明它可以结合smpdl3b并保护 足细胞的形态，C57BL/6小鼠的SD RT诱导RN减少，但在我们新发达的条件中没有 Podocyte特异性SMPDL3B敲除小鼠。基于这些数据，我们假设鞘脂起着至关重要的 控制RN的辐射诱导的足细胞病。 目的是研究SMPDL3B在低剂量分离后肾脏损伤中的机理作用 放射疗法（F-RT）与并发顺铂（CDDP），因为这代表了许多固体的护理标准 癌症。我们的长期目标是发现基于分子的RN的保护性或缓解策略，以及 潜在的化学疗法诱导的肾毒性。我们将通过以下三个特定的三个特定来检验我们的假设 目的使用合并的体内体外方法： 目标1：确定SMPDL3B是否调节与RT相关肾脏损伤和功能的严重性和潜伏期 在临床上与F-RT，CDDP和并发F-RT+CDDP之后的RN。这个目标还将探讨 使用C57BL/6小鼠和我们独特的SMPDL3B-KNOCKOUT 和SMPDL3B诱导的鼠标模型。 目的2：确定SMPDL3B的足细胞表达影响RT介导的Podocyte的机制 和肾小球内皮细胞（GEC）损伤。我们假设SMPDL3B会影响RT诱导 足细胞Ezrin的隔室化，并通过改变的内皮素-1（EDN1）和END1受体影响GEC A型（Ednra）串扰。将通过与缺乏足细胞共同培养的GEC来研究RT后的GEC生存 或表达SMPDL3B。 目标3：确定靶向鞘脂是否阻止RN。我们将调查SMPDL3B或S1P的保护 F-RT，CDDP和F-RT+CDDP之后，C57BL/6小鼠的长期功能性肾损伤是否会避免长期功能性损伤。机制 将使用我们独特的SMPDL3B-KNOCKOUT和SMPDL3B诱导的鼠标模型来确认。 这些研究的发现将很重要，因为它们为分子靶向的潜力 RT和辐射相关的肾脏损伤后，RT缓解措施，并在RT和组合损伤后进行缓解。

项目成果

Detection and Quantification of Lipid Droplets in Differentiated Human Podocytes.

分化的人足细胞中脂滴的检测和定量。

• DOI: 10.1007/978-1-4939-9488-5_17
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Mallela,ShamroopKumar;Patel,DevangMaheshkumar;Ducasa,GloriaMichelle;Merscher,Sandra;Fornoni,Alessia;Al-Ali,Hassan
• 通讯作者: Al-Ali,Hassan

Role of Sphingolipid Signaling in Glomerular Diseases: Focus on DKD and FSGS.

• DOI: 10.33696/signaling.1.013
• 发表时间: 2020-09-01
• 期刊: Journal of cellular signaling
• 作者: Mitrofanova, Alla;Drexler, Yelena;Fornoni, Alessia
• 通讯作者: Fornoni, Alessia

Noninvasive assessment of radiation-induced renal injury in mice.

• DOI: 10.1080/09553002.2021.1876950
• 发表时间: 2021
• 期刊: International journal of radiation biology
• 影响因子: 2.6
• 作者: Ahmad A;Shi J;Ansari S;Afaghani J;Molina J;Pollack A;Merscher S;Zeidan YH;Fornoni A;Marples B
• 通讯作者: Marples B

Sphingomyelin phosphodiesterase acid like 3B (SMPDL3b) regulates Perilipin5 (PLIN5) expression and mediates lipid droplet formation.

• DOI: 10.1016/j.gendis.2021.12.014
• 发表时间: 2022-11
• 期刊: Genes & diseases
• 影响因子: 6.8
• 作者: Mallela SK;Ge M;Molina J;Santos JV;Kim JJ;Mitrofanova A;Al-Ali H;Marples B;Merscher S;Fornoni A
• 通讯作者: Fornoni A

Radiation nephropathy: Mechanisms of injury and recovery in a murine model.

• DOI: 10.1016/j.radonc.2023.109813
• 发表时间: 2023-07
• 期刊: Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology
• 作者: Anis Ahmad;Junwei Shi;S. Ansari;S. Merscher;Alana Pollack;Y. Zeidan;A. Fornoni;B. Marples