Modulation of vascular endothelial alloimmunogenicity by nanoparticle-mediated ge

纳米颗粒介导的基因调节血管内皮同种免疫原性

基本信息

批准号：
8718161
负责人：
PARWIZ ABRAHIMI
金额：
$ 2.73万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-16 至 2017-08-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8718161
关键词：
Acids Adoption Adverse effects Alleles Allogenic Attenuated Basic Science Blood Vessels CD4 Positive T Lymphocytes CD8B1 gene Caring Cell Line Cells Clinical Coupled Dendritic Cells Disease Dose Drug Targeting Drug toxicity Encapsulated Endothelial Cells Endothelium Enzyme-Linked Immunosorbent Assay FDA approved Failure Foundations Frequencies Graft Rejection Health Heart failure Histocompatibility Antigens Class II Human Immune Immune response Immune system Immunofluorescence Immunologic Immunology Immunosuppression Immunosuppressive Agents Implant In Vitro Infiltration Inflammation Injury Interferons Kidney Label Length Leukocytes Life Ligands Link Lipids Liver Living Donors Lung Lymphocyte Lymphoid MHC class II transactivator protein Major Histocompatibility Complex Malignant Neoplasms Measures Mediating Memory Metabolic Modeling Operative Surgical Procedures Opportunistic Infections Organ Organ Transplantation Outcome Pathologic Patients Perfusion Peripheral Blood Mononuclear Cell Phenotype Physicians Polymers Postoperative Period Process Production Proteins Reaction Regimen Reperfusion Injury Reporting Resistance Risk Rodent Model Scientist Secondary to Shapes Small Interfering RNA Solid Staging Staining method Stains System T cell response T memory cell T-Cell Activation T-Lymphocyte Techniques Therapeutic Therapeutic immunosuppression Time Toxic effect Training Transcript Transfection Transplant Recipients Transplantation Vascular Endothelial Cell base career cell type clinically relevant cytokine design ex vivo perfusion functional outcomes graft function immunogenicity in vivo knock-down morphometry mouse model nanoparticle novel pre-clinical public health relevance research study response success therapy design transcription factor transplantation medicine

项目摘要

DESCRIPTION (provided by applicant): Description: Solid organ transplantation is the only curative treatment for end-stage kidney, liver, lung and heart failure but its success is limited b immunological rejection. Although immunosuppressive therapies are increasingly effective, their value is limited due to memory cell resistance to suppression, toxicities, and complications secondary to the general suppression of the host immune system, including opportunistic infections and malignancies. Rejection is initiated in humans when graft cells that express class I and class II MHC molecules, in particular endothelial cells that line the vasculature of the organ, are directly recognized by alloreactive memory CD8+ and CD4+ T lymphocytes, respectively. This project seeks to reduce rejection by altering the transplanted organ during the period around the time of transplant surgery when the host immune response is being shaped. I hypothesize that reducing MHC class II on graft endothelium will be sufficient to attenuate rejection responses by limiting the activation of CD4+ T cells, the principal orchestrators of graf rejection. I propose to accomplish this by using biodegradable nanoparticles to transfect graft endothelial cells ex vivo and deliver small interfering RNA (siRNA) that suppress the expression of the class II transactivator (CIITA), the master regulator of MHC class II expression. My specific aims are: (1) to characterize the effects of reducing MHC class II expression on cultured ECs by lipid-mediated transfection of anti-CIITA siRNA on modulating CD4+ and CD8+ alloresponses, as measured by cytokine production, proliferation and adoption of pathogenic phenotypes in vitro; (2) to characterize and optimize a nanoparticle delivery system that encapsulates anti-CIITA siRNA and is targeted to cultured human ECs by different ligands with an emphasis on maximizing the magnitude and duration of knockdown without inducing toxicity; and (3) to use optimized EC-targeted nanoparticles to transfect EC lining intact segments of human vessels and to assess the modulation of rejection of such vessels by allogeneic human peripheral blood mononuclear cells in a pre- clinical humanized mouse model.

描述（由申请人提供）：描述：固体器官移植是终末期肾脏，肝脏，肺和心力衰竭的唯一治疗方法，但其成功是有限的B免疫抑制。尽管免疫抑制疗法越来越有效，但由于记忆细胞对抑制，毒性和并发症的耐药性，其价值受到限制。当表达I类和II类MHC分子的移植细胞，特别是在器官的脉管系统中的内皮细胞，分别通过同种反应性记忆CD8+和CD4+ T型淋巴细胞直接识别。该项目旨在通过改变移植手术时发生宿主免疫反应时改变移植器官来减少排斥。我假设在移植内皮上减少MHC II类的II将足以通过限制CD4+ T细胞的激活来减轻排斥反应，这是GRAF排斥的主要编排者。我建议通过使用可生物降解的纳米颗粒将移植物内皮细胞移动，并提供小型干扰RNA（siRNA），从而抑制了MHC II类表达的主要调节剂II类反式激活剂（CIITA）的表达。我的具体目的是：（1）表征通过脂质介导的抗CIITA siRNA转染降低MHC II类表达对培养的EC的影响，对通过细胞因子的产生，PROPRISINION，PATHENIC MENENEGINED型号测量了调节CD4+和CD8+同种异酶体外; （2）表征和优化纳米颗粒递送系统，该系统封装了抗CIITA siRNA，并通过不同的配体针对培养的人类EC，重点是最大程度地提高敲低的幅度和术语而不诱导毒性；（3）使用优化的EC靶向纳米颗粒来转染人体血管的完整段，并评估在临床人性化的小鼠模型中，通过同种异体人类外周血单核细胞对这种血管的抑制调节。