Lipid mediated oral tolerance

脂质介导的口服耐受

基本信息

批准号：
10416195
负责人：
SATHY VENKAT BALU-IYER
金额：
$ 46.55万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-16 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10416195
关键词：
Address Adopted Adoptive Transfer Animal Model Antibody Response Antigen-Presenting Cells Antigens Autoimmune Diseases Autoimmunity B-Lymphocytes Biological Biophysics Blocking Antibodies Cells Charge Clinical Data Dendritic Cells Development Factor VIII Food Hypersensitivity Foundations Generations Glucan 1,4-alpha-Glucosidase Glycogen storage disease type II Goals Gut associated lymphoid tissue Head Health Hemophilia A Human Hypersensitivity Imaging Techniques Immune Immune Tolerance Immune response Immunologics Immunotherapy In Vitro Interleukin-10 Knockout Mice Knowledge Label Lecithin Length Life Lipids M cell Mediating Medical Methodology Mission Modeling Molecular Mus Oral Outcome Ovalbumin Particulate Phosphatidylserines Property Proteins Public Health Publishing Receptor Signaling Regulatory T-Lymphocyte Research Role Savings Signal Transduction Structure Surface System Techniques Testing Transgenic Mice United States National Institutes of Health Work base biophysical properties biophysical techniques density design immunogenicity in vivo innovation interdisciplinary approach lipid nanoparticle lipid structure lysophosphatidylserine mouse model nanoparticle nanoparticle exposure novel oral tolerance particle prevent receptor receptor binding response therapeutic protein tool trafficking uptake

项目摘要

Project Summary: Autoimmune conditions, allergies and immunogenicity against therapeutic proteins are caused by unwanted immune responses against proteins. Our long-term goal is to rationally develop an immunotherapy platform using protein delivery strategies to treat these clinical conditions. We observed that an oral pre-administration of antigens loaded in lyso-phosphatidylserine (lysoPS), but not in double chain PS, based nanoparticles effectively reduced unwanted immune response against several protein antigens, forming the basis for an innovative immunotherapy. The overall objective of this application is to elucidate the cellular and molecular mechanisms of oral delivery and tolerance mediated by lysoPS based nanoparticles. The rationale for the proposed research is that once we gain a mechanistic understanding of the tolerogenic property of lysoPS, it would result in an innovative immunotherapy approach and clinical strategies to prevent and reverse immunogenicity of therapeutic proteins and to treat/cure autoimmune conditions and allergies. We also found that there are differences in biophysical characteristics such as PS exposure of the nanoparticles comprised of lyso and double chain PS. Based on these observations, our working hypothesis is that lysoPS mediated tolerance is due to its distinct structural/biophysical characteristics that influences its access to gut associated lymphoid tissue, receptor mediated cellular uptake/signaling and underlying molecular and cellular mechanism of oral tolerance. Three specific aims (SA) are proposed to test our hypothesis and accomplish our objective using a multidisciplinary approach. In SA 1, we will investigate the impact of biophysical properties of lysoPS based lipidic particles and its impact on M-cell mediated particulate uptake. Biophysical characteristics such as PS exposure, clustering, and lamellar properties will be investigated using orthogonal techniques. We will adopt in vitro, ex vivo and in vivo studies using M-cell deficient mice to investigate the impact of M-cell engagement of PS based particles containing model antigen Ovalbumin OVA. In SA 2, we will investigate the impact of PS exposure on receptor binding and intracellular signaling. Based on our observation that functional blocking antibody against Tim-4 receptor reversed PS mediated tolerance, we will investigate the role of this receptor binding in tolerance using advanced cell-biological and imaging techniques and also using knock-out mice models. In SA 3, we will investigate the mechanism of lysoPS mediated oral tolerance towards OVA, in particular, the effects on gut resident dendritic cells, generation of regulatory T-cells and B-cell response. The generation of Tregs will be investigated using OT-II transgenic mice and by adoptive transfer approaches. We will investigate the impact of Tregs on B-cells by investigating the impact of lysoPS on the expression of IL-10, TGFb and neuritin. Overall, our combined approach investigating the biophysical, cell biological, innate and adaptive mechanisms of lysoPS tolerance will lay the foundation for a comprehensive clinical approach to reduce immunogenicity of therapeutic proteins as well as prevent and reverse autoimmunity and allergies.

项目摘要：针对治疗蛋白的自身免疫性条件，过敏和免疫原性是由对蛋白质的免疫反应引起的。我们的长期目标是合理发展免疫疗法平台使用蛋白质输送策略来治疗这些临床状况。我们观察到一个在溶菌 - 磷脂酰丝氨酸（Lysops）中加载的抗原的口服预先给药，而不是在双链PS中，基于双链PS 纳米颗粒有效地减少了针对几种蛋白质抗原的不需要的免疫反应，形成了创新免疫疗法的基础。该应用的总体目的是阐明细胞和基于Lysops的纳米颗粒介导的口服递送和耐受性的分子机制。理由拟议的研究是，一旦我们对lysops的耐受性特性有机械理解，这将导致一种创新的免疫疗法方法和临床策略，以预防和逆转治疗蛋白的免疫原性，并治疗/治疗自身免疫性和过敏。我们还发现生物物理特征存在差异，例如由纳米颗粒的PS暴露 Lyso和Double Chain PS。基于这些观察结果，我们的工作假设是Lysops介导耐受性是由于其独特的结构/生物物理特征会影响其进入肠道的访问淋巴组织，受体介导的细胞摄取/信号传导以及基础分子和细胞机制口服耐受性。提出了三个具体目标（SA）来检验我们的假设并实现我们的目标使用多学科方法。在SA 1中，我们将研究Lysops生物物理特性的影响基于脂质颗粒及其对M细胞介导的颗粒摄取的影响。生物物理特征，例如 PS暴露，聚类和层状特性将使用正交技术研究。我们将采用体外，体内和体内研究使用M细胞缺乏小鼠研究M细胞参与的影响含有模型抗原椭圆蛋白OVA的基于PS的颗粒。在SA 2中，我们将研究PS的影响受体结合和细胞内信号传导的暴露。根据我们的观察到功能阻挡针对TIM-4受体的抗体逆转PS介导的耐受性，我们将研究该受体的作用使用高级细胞生物学和成像技术在耐受性中结合，还使用敲除小鼠型号。在SA 3中，我们将研究Lysops介导的口服耐受性的机理，特别是对肠道常驻树突状细胞的影响，调节性T细胞的产生和B细胞反应。这一代 Treg将使用OT-II转基因小鼠和收养转移方法研究。我们将调查通过研究Lysops对IL-10，TGFB和TGFB的表达的影响，Treg对B细胞的影响神经蛋白。总体而言，我们研究生物物理，细胞生物学，先天和适应性的合并方法 Lysops耐受性的机制将为综合临床方法奠定基础治疗蛋白的免疫原性以及预防和反向自身免疫性和过敏。