Synthetic mRNA-mediated reversible immunocontraception

合成 mRNA 介导的可逆免疫避孕

• 批准号: 10018523
• 负责人: Deborah J Anderson
• 金额: $ 38.05万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-13 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10018523
• 关键词: Address; Aerosols; Affect; Antibodies; Antibody Formation; Antigens; Applications Grants; Architecture; Back; Biological; Breast Cancer Risk Factor; Cells; Columnar Epithelium; Contraceptive Agents; Contraceptive methods; Development; Dose; Drug Kinetics; Epithelial; Epithelium; Estrogen receptor positive; Female; Female Contraceptive Agents; Fertility; Goals; HIV; Half-Life; Hormonal; Human; Human Herpesvirus 2; IgG1; Immunoglobulin A; Immunoglobulin G; Immunologic Contraception; In Vitro; Inflammation; Lesion; Macaca; Macaca mulatta; Mediating; Messenger RNA; Methods; Modeling; Molecular; Monoclonal Antibodies; Mucous Membrane; Mucous body substance; Phase; Pregnancy; Quality of life; Reproductive Biology; Reproductive Health; Safety; Sexually Transmitted Diseases; Sheep; Site; Spermatozoa antibody; Techniques; Testing; Tissues; Vagina; Water; Woman; base; cell type; cost effective; design; experimental study; in vivo; inhibiting antibody; innovation; mRNA delivery; migration; novel strategies; pre-clinical; reproductive hormone; reproductive tract; sperm cell; sperm function; unintended pregnancy

Project summary Currently 72% of women who practice contraception use hormonal methods, but there is frequent dissatisfaction with these methods, due to quality of life and safety concerns. Therefore, there is a clear need for new approaches to non-hormonal female contraceptives that are easy to use, woman-applied, and have a controllable duration of action. Reversible immunocontraception offers a non-hormonal solution, where anti- sperm antibodies are introduced into the female reproductive tract (FRT) and inhibit sperm function. This approach, though, has a number of challenges including: discovery of a specific and effective monoclonal antibody (mAb) against a human sperm antigen, and a safe and reliable method for introduction of mAbs that is temporally and spatially controllable. For this grant application, reproductive biology expertise and a molecular toolbox has been brought together to overcome these challenges. An anti-sperm antibody has been identified with well-characterized mechanisms of action that impact fertility, and an innovative method has been identified to deliver the antibody to the FRT. Here a synthetic mRNA-based approach is proposed to deliver a sperm agglutinating and mucus trapping antibody to the FRT. To date, the ability to express high levels of antibody for over 28 days with a single administration of mRNA in the FRT of sheep has been demonstrated. Delivery is achieved through direct, rapid, aerosol exposure of the FRT epithelium to naked mRNA in water. Persistence of the antibody in secretions was achieved through the incorporation of a GPI-linker into the heavy chain of the antibody. This combination allows for rapid delivery and expression, and for controllable persistence of the protective mAb in secretions. In the R61 phase, the mechanism of delivery under relevant conditions will be optimized and explored; antibody design questions will also be addressed that will allow for tunable persistence of the antibody in mucus secretions. In the R33 phase, pharmacokinetic experiments will be performed in macaques, as well as continued antibody optimization. Last, sperm challenge experiments will be performed in vivo to demonstrate efficacy. The long-term goal is to develop a cost-effective mRNA-based approach for expressing anti-sperm antibodies in the FRT. The short-term goals are to optimize the approach, answer mechanistic questions, and demonstrate pre-clinical feasibility in the macaque model. If successful, a new paradigm for contraception will be demonstrated, one that can be combined with the co-expression of anti-STI antibodies, such as those against HIV and HSV-2.

项目概要 目前72%的女性避孕方法采用激素避孕法，但经常出现不满意的情况 由于生活质量和安全问题，使用这些方法。因此，显然需要新的 易于使用、女性使用且具有良好效果的非激素女性避孕方法 可控的作用持续时间。可逆免疫避孕提供了一种非激素解决方案，其中抗 精子抗体被引入女性生殖道（FRT）并抑制精子功能。这 然而，该方法面临许多挑战，包括：发现一种特定且有效的单克隆抗体 针对人类精子抗原的抗体（mAb），以及引入mAb的安全可靠的方法 时间和空间可控。对于这项拨款申请，生殖生物学专业知识和分子 工具箱已汇集在一起​​以克服这些挑战。已鉴定出抗精子抗体 具有影响生育能力的明确作用机制，并且已经确定了一种创新方法 将抗体递送至 FRT。这里提出了一种基于合成 mRNA 的方法来输送精子 FRT 凝集和粘液捕获抗体。迄今为止，表达高水平抗体的能力 已经证明，在绵羊 FRT 中单次施用 mRNA 可以达到超过 28 天的效果。交货是 通过将 FRT 上皮直接、快速、气溶胶暴露于水中裸露的 mRNA 来实现。坚持 抗体在分泌物中的表达是通过将 GPI 连接子掺入抗体的重链来实现的。 抗体。这种组合允许快速递送和表达，以及可控的持久性 分泌物中的保护性单克隆抗体。在R61阶段，相关条件下的交付机制将是 优化和探索；抗体设计问题也将得到解决，这将允许可调的持久性 粘液分泌物中的抗体。在R33阶段，将进行药代动力学实验 猕猴，以及持续的抗体优化。最后，精子挑战实验将在 体内证明功效。长期目标是开发一种具有成本效益的基于 mRNA 的方法 在 FRT 中表达抗精子抗体。短期目标是优化方法，回答 机制问题，并在猕猴模型中证明临床前可行性。如果成功的话，会出现一个新的 将展示一种可以与抗性传播感染共表达相结合的避孕范例 抗体，例如抗 HIV 和 HSV-2 的抗体。