Lipid mediated oral tolerance

脂质介导的口服耐受

• 批准号: 10647679
• 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
• 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/10647679
• 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; 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; Pharmaceutical Preparations; Phosphatidylserines; Property; Proteins; Public Health; Publishing; Regulatory T-Lymphocyte; Research; Role; 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

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：自身免疫性疾病、过敏和针对治疗性蛋白质的免疫原性是 由针对蛋白质的不必要的免疫反应引起。我们的长远目标是合理发展 使用蛋白质递送策略来治疗这些临床病症的免疫治疗平台。我们观察到， 口服预先施用负载在溶血磷脂酰丝氨酸（lysoPS）中但不负载在双链PS中的抗原， 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应答，形成了 创新免疫疗法的基础。本申请的总体目标是阐明细胞和细胞外基质中的蛋白质。 基于lysoPS的纳米颗粒介导的口服递送和耐受性的分子机制。的理由 所提出的研究是一旦我们获得了对lysoPS的致耐受性性质的机理理解， 它将产生一种创新的免疫治疗方法和预防和逆转的临床策略 治疗性蛋白质的免疫原性和治疗/治愈自身免疫性病症和过敏。我们还发现 在生物物理特性上存在差异，例如由以下组成的纳米颗粒的PS暴露 lyso和双链PS。基于这些观察，我们的工作假设是lysoPS介导的 耐受性是由于其独特的结构/生物物理特征，其影响其进入肠道相关组织的能力。 淋巴组织、受体介导的细胞摄取/信号传导以及潜在的分子和细胞机制 口服耐受性。提出了三个具体目标（SA）来验证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的生物物理特性对 基于脂质颗粒的脂质颗粒及其对M细胞介导的颗粒摄取的影响。生物物理特征，如 PS曝光，聚类，和层状性能将使用正交技术进行研究。我们将采取 使用M细胞缺陷小鼠进行体外、离体和体内研究，以研究 含有模型抗原卵清蛋白OVA的基于PS的颗粒。在SA 2中，我们将研究PS的影响 暴露于受体结合和细胞内信号传导。根据我们的观察，功能性阻滞 针对Tim-4受体的抗体逆转PS介导的耐受，我们将研究该受体的作用 使用先进的细胞生物学和成像技术以及敲除小鼠 模型在SA 3中，我们将研究溶血PS介导的对OVA的口服耐受的机制，特别是， 对肠道树突状细胞、调节性T细胞和B细胞反应的影响。生成 将使用OT-II转基因小鼠和通过过继转移方法研究TcB的表达。我们将调查 通过研究lysoPS对IL-10、TGF β和TNF α表达的影响， 神经素。总的来说，我们研究生物物理学，细胞生物学，先天和适应性的综合方法 lysoPS耐受机制将为全面的临床方法奠定基础， 治疗性蛋白质免疫原性以及预防和逆转自身免疫和变态反应。