Encapsulated Active Vitamin D Vaccine for the Treatment of Multiple Sclerosis

用于治疗多发性硬化症的封装活性维生素 D 疫苗

• 批准号: 8313902
• 负责人: Kristy M Ainslie
• 金额: $ 19.06万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-15 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8313902
• 关键词: Acetone; Acids; Adoptive Transfer; Adverse effects; Affect; Alcohols; American; Antigens; Autoimmune Diseases; Autoimmune Process; Bone Marrow; CD4 Positive T Lymphocytes; CD8B1 gene; Calcitriol; Cell Culture Techniques; Cells; Clinical; Coculture Techniques; Data; Dendritic Cells; Development; Dexamethasone; Dextrans; Disease; Disease Progression; Dose; Emulsions; Encapsulated; Environment; Evaluation; Experimental Autoimmune Encephalomyelitis; Figs - dietary; Flow Cytometry; Freund' s Adjuvant; Glycolates; Goals; Grant; Image; Immune; Immune response; Immune system; Immunization; Immunosuppression; In Vitro; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Lethal Dose 50; Lipopolysaccharides; Lysosomes; Measurement; Measures; Mediating; Metabolic; Methods; Modeling; Multiple Sclerosis; Mus; Myelin Basic Proteins; Nanotechnology; Outcome; Particle Size; Pathogenesis; Patients; Peptides; Phagocytes; Phagocytosis; Phagosomes; Pharmaceutical Preparations; Polymers; Population; Proteins; Rheumatoid Arthritis; Surface; System; Techniques; Technology; Testing; Tissues; Vaccines; Vitamin D; calcification; central nervous system demyelinating disorder; comparative efficacy; cytokine; design; dextran; in vivo; innovation; nano; nanoforms; nanoparticle; novel; particle; prevent; response

DESCRIPTION (provided by applicant): We present a translational method for developing a tolerogenic vaccine to drastically decrease the burden of patients with multiple sclerosis (MS) by using nanotechnology. We hypothesize that nano/microparticles fabricated from the polymer acetalated dextran (Ac-DEX) that encapsulate the active form of vitamin D, and myelin basic protein (MBP) will initiate the formation of MBP specific inducible T-reg cells (iT-regs) capable of suppressing MS in an antigen specific manner. Our project is innovative because: 1.) Passive in vivo targeting: Previous methods to develop tolerogenic DCs to treatment MS require ex vivo manipulation. Phagocytic dendritic cells (DCs) allow passive targeting since the nanoparticles are too large to be taken up by non-phagocytes. Passive targeting leads to in vivo delivery of tolerogenic agents to the DCs, such as the active form of vitamin D (calcitriol). Currently, calcitriol is ineffective at treating MS because at the concentrations necessary to ameliorate the disease, whole tissue calcification is prominent. 2.) Dose sparing: Our preliminary studies indicate that encapsulation of immunomodulatory agents leads to dose sparing. We hypothesize that we will be able to target DCs in vivo and deliver payload intracellularly, thus decreasing the amount of calcitriol needed, thereby preventing tissue calcification. The targeted DCs will become tolerogenic, which will then be capable of inducing the formation of MS suppressing iT-reg cells. 3.) Acid sensitive nanoparticles: Acid sensitive nanoparticles exposed to the acidic environment of the phagocyte lysosome will degrade, releasing the encapsulated antigen and initiating an immune response. We have previously shown that using our pH sensitive polymer Ac-Dex, we can drastically increase both CD4 and CD8 presentation compared to free protein, and protein encapsulated in other common polymeric carriers. 4.) Specific immune dampening: Current methods of MS treatment require non-specific systemic immune suppression. We present a method in which we can generate a MBP specific iT-reg population that can suppress against MS without suppressing the entire immune system. We present preliminary data that shows encapsulation of dexamethasone and MBP peptide results in increased formation of MBP specific iT-regs in the CNS and reduced disease progression with treatment before experimental autoimmune encephalomyelitis (EAE) onset in mice. In order to develop these nanoparticles and test their efficacy we present the following three specific aims: Specific Aim 1 will focus on calcitriol and MBP peptide encapsulation in nanoparticles. Specific Aim 2 will focus on evaluating calcitriol encapsulated nanoparticles in inducing tolerogenic DC and iT-regs in vitro. Specific Aim 3 will be testing the formation of iTregs in vivo by the immunization with co-encapsulated MBP and calcitriol. Finally, by using the EAE model of MS, we will treat mice with our nanoparticles. If successful, we will take the data generated in this grant and apply for an R01. In addition, our nanoparticle system could be applied to other autoimmune diseases such as Type 1 diabetes and rheumatoid arthritis.

描述（由申请人提供）：我们提出了一种利用纳米技术开发耐受性疫苗的转化方法，以大幅减轻多发性硬化症（MS）患者的负担。我们假设，由包裹维生素 D 活性形式的聚合物乙酰化右旋糖酐 (Ac-DEX) 和髓磷脂碱性蛋白 (MBP) 制成的纳米/微粒将引发 MBP 特异性诱导性 T-reg 细胞 (iT-regs) 的形成，该细胞能够以抗原特异性方式抑制 MS。我们的项目具有创新性，因为： 1.) 被动体内靶向：以前开发耐受性 DC 来治疗 MS 的方法需要离体操作。吞噬树突状细胞（DC）允许被动靶向，因为纳米颗粒太大而不能被非吞噬细胞吸收。被动靶向导致体内向 DC 递送耐受剂，例如活性形式的维生素 D（骨化三醇）。目前，骨化三醇对于治疗多发性硬化症无效，因为在改善疾病所需的浓度下，整个组织钙化是显着的。 2.) 剂量节省：我们的初步研究表明，免疫调节剂的封装可以节省剂量。我们假设我们将能够在体内靶向树突状细胞并在细胞内递送有效负载，从而减少所需的骨化三醇的量，从而防止组织钙化。目标 DC 将变得耐受，然后能够诱导 MS 抑制 iT-reg 细胞的形成。 3.)酸敏感纳米颗粒：酸敏感纳米颗粒暴露于吞噬细胞溶酶体的酸性环境中会降解，释放出封装的抗原并引发免疫反应。我们之前已经证明，与游离蛋白质和封装在其他常见聚合物载体中的蛋白质相比，使用 pH 敏感聚合物 Ac-Dex 可以显着增加 CD4 和 CD8 的表达。 4.) 特异性免疫抑制：目前治疗多发性硬化症的方法需要非特异性全身免疫抑制。我们提出了一种方法，可以生成 MBP 特异性 iT-reg 群体，该群体可以抑制 MS，而不抑制整个免疫系统。我们提供的初步数据显示，地塞米松和 MBP 肽的封装导致中枢神经系统中 MBP 特异性 iT-reg 的形成增加，并在小鼠实验性自身免疫性脑脊髓炎 (EAE) 发病前进行治疗，从而减少疾病进展。为了开发这些纳米颗粒并测试其功效，我们提出了以下三个具体目标： 具体目标 1 将重点关注纳米颗粒中骨化三醇和 MBP 肽的封装。具体目标 2 将重点评估骨化三醇封装的纳米颗粒在体外诱导耐受性 DC 和 iT-reg 方面的作用。具体目标 3 将通过共封装 MBP 和骨化三醇进行免疫来测试体内 iTreg 的形成。最后，通过使用 MS 的 EAE 模型，我们将用我们的纳米颗粒治疗小鼠。如果成功，我们将利用这笔赠款中生成的数据并申请 R01。此外，我们的纳米颗粒系统还可应用于其他自身免疫性疾病，如 1 型糖尿病和类风湿性关节炎。

项目成果

Treatment of experimental autoimmune encephalomyelitis by codelivery of disease associated Peptide and dexamethasone in acetalated dextran microparticles.

• DOI: 10.1021/mp4005172
• 发表时间: 2014-03-03
• 期刊: Molecular pharmaceutics
• 影响因子: 4.9
• 作者: Peine KJ;Guerau-de-Arellano M;Lee P;Kanthamneni N;Severin M;Probst GD;Peng H;Yang Y;Vangundy Z;Papenfuss TL;Lovett-Racke AE;Bachelder EM;Ainslie KM
• 通讯作者: Ainslie KM