Delivery of passive immunity against HIV using commensal vaginal Lactobacillus bioengineered to secrete broadly neutralizing domain antibodies

使用经生物工程改造可分泌广泛中和域抗体的共生阴道乳杆菌提供针对 HIV 的被动免疫

• 批准号: 8906724
• 负责人: Laurel A. Lagenaur
• 金额: $ 138.85万
• 依托单位: OSEL, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8906724
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Acute; Address; Antibiotic susceptibility; Antibodies; Biochemical; Biological; Biological Assay; Biological Response Modifier Therapy; Biomedical Engineering; Cervical; Chromosomal Insertion; Clinical; Codon Nucleotides; Development; Drug Formulations; Engineering; Enzyme-Linked Immunosorbent Assay; Epidemic; Epidemiologic Studies; Epitopes; Female; Female of child bearing age; Freeze Drying; Future; Genetic; Genomics; Goals; HIV; HIV Infections; HIV-1; Health; Heterosexuals; Human; Infection; Intravaginal Administration; Investigational Drugs; Lactobacillus; Life; Macaca; Macaca mulatta; Manufacturer Name; Measures; Modeling; Mucous Membrane; Mus; Oryctolagus cuniculus; Passive Immunity; Pharmacologic Substance; Phase; Principal Investigator; Process; Production; Property; Protective Agents; Proteins; Protocols documentation; Recombinants; Research Design; Resistance profile; Risk; Sexually Transmitted Diseases; Site; Small Business Innovation Research Grant; Surface; Tablets; Technology; Testing; Toxic effect; Vagina; Viral; Virus; Virus Diseases; Woman; cost effective; efficacy testing; inhibitor/antagonist; integration site; irritation; manufacturing process development; method development; microbicide; nonhuman primate; novel; pathogen; phase 1 study; phase 2 study; pre-clinical; preclinical efficacy; preclinical safety; prevent; programs; promoter; rectal; safety study; safety testing; scale up; self-renewal; simian human immunodeficiency virus; social; transmission process; vaginal lactobacilli

DESCRIPTION: Women bear the brunt of the global AIDS epidemic due in part to their inherent social and biological vulnerability. Biologically, the cervical and vaginal mucosae are vulnerable targets for HIV transmission. In healthy women of childbearing age, mucosal antibodies and microbiota, including Lactobacillus jensenii, help protect these surfaces and serve to block the colonization and infection by intruding pathogens. Epidemiological studies suggest that the loss of vaginal lactobacilli is associated with an increased risk of heterosexual HIV-1 transmission and other sexually transmitted infections. Osel, Inc. has used the properties of these natural protective agents by engineering a native human vaginal Lactobacillus , L. jensenii 1153, to express potent and broadly neutralizing single domain antibodies directed against HIV, termed MucoCept-Ab. Osel recently showed proof-of-concept for this recombinant Lactobacillus approach, by expressing an HIV-1 entry inhibitor mCV-N, from L. jensenii, which reduced SHIV virus transmission in macaques by 63% (Lagenaur et al., Mucosal Immunol, (Jul 6, 2011)). Phase II studies will advance MucoCept-Ab through preclinical development. These studies will focus on developing a safe, efficacious, and stable MucoCept-Ab product, and will include strain optimization, formulation, process development, manufacturing, preclinical safety testing, and efficacy testing in a nonhuman primate (NHP) viral challenge model. The goal of this Phase II is to develop a Lactobacillus strain capable of delivering an antibody locally to prevent HIV infection at the vaginal or rectal mucosa. This strain will be formulated into a cost-effective product capable of persistent protection that is coitally-independent and controlled by women. Aim 1: MucoCept-Ab API optimization and characterization: A) Complete MucoCept-Ab strain optimization: maximize promoter strength, optimize codons for L. jensenii, remove epitope tag, and test two genomic integration sites; B) Complete microbiological characterization of the MucoCept-Ab strain; genetic stability, sequence chromosomal insertion site of expression cassette, biochemical profiling, antibiotic susceptibility/resistant profiling; AP-m36.4 expressio levels; C) Establish ability of the MucoCept-Ab strain to vaginally colonize female rhesus macaques. Aim 2: MucoCept-Ab method development: A) Develop antibodies and ELISA to detect and measure AP-m36.4 levels; B) Develop qPCR assay to measure levels of the MucoCept-Ab strain. Aim 3: MucoCept-Ab formulation, process development, and manufacturing: A) Develop formulation and lyophilization process for production of a stable MucoCept-Ab product; B) Transfer MucoCept-Ab strain and technology to GMP manufacturer for production of MCB and WCB, scale up process development, and production of MucoCept-Ab for preclinical safety studies. Aim 4: MucoCept-Ab preclinical safety and efficacy: A) Evaluate acute toxicity and systemic exposure of the MucoCept-Ab strain and purified AP-m36.4 protein following vaginal administration in mice; B) Evaluate acute toxicity of formulated MucoCept-Ab in the rabbit vaginal irritation model; C) Evaluate subchronic toxicity, vaginal colonization, and AP-m36.4 levels following vaginal administration of the MucoCept-Ab product in female rhesus macaques; D) Evaluate efficacy of the MucoCept-Ab product in female rhesus macaques using a repeated vaginal SHIV challenge protocol. If successful, this approach could provide a safe, yet inexpensive means to deliver passive immunity against HIV to the vaginal mucosa and to address the urgent need for female-controlled approaches to prevent sexually transmitted viral infection.

描述：妇女在全球艾滋病流行中首当其冲，部分原因是她们固有的社会和生物脆弱性。从生物学角度来说，宫颈和阴道粘膜是艾滋病毒传播的脆弱目标。在健康的育龄妇女中，粘膜抗体和微生物群（包括詹氏乳杆菌）有助于保护这些表面，并有助于阻止入侵病原体的定植和感染。流行病学研究表明，阴道乳酸杆菌的丧失与异性传播 HIV-1 和其他性传播感染的风险增加有关。 Osel, Inc. 利用这些天然保护剂的特性，通过改造天然人阴道乳杆菌（L. jensenii 1153）来表达针对 HIV 的有效且广泛中和的单域抗体，称为 MucoCept-Ab。 Osel 最近展示了这种重组乳杆菌方法的概念验证，通过表达来自詹氏乳杆菌的 HIV-1 进入抑制剂 mCV-N，可将猕猴中的 SHIV 病毒传播减少 63%（Lagenaur 等人，Mucosal Immunol，（2011 年 7 月 6 日））。 II 期研究将通过临床前开发推进 MucoCept-Ab 的发展。这些研究将重点开发安全、有效和稳定的 MucoCept-Ab 产品，并将包括菌株优化、配方、工艺开发、制造、临床前安全性测试以及非人类灵长类动物 (NHP) 病毒攻击模型中的功效测试。第二阶段的目标是开发一种能够局部递送抗体以预防阴道或直肠粘膜的艾滋病毒感染的乳杆菌菌株。该菌株将被配制为具有成本效益的产品，能够持续保护性交独立且由女性控制。目标 1：MucoCept-Ab API 优化和表征：A) 完成 MucoCept-Ab 菌株优化：最大化启动子强度、优化 L. jensenii 密码子、去除表位标签并测试两个基因组整合位点； B) MucoCept-Ab 菌株的完整微生物学特征；遗传稳定性、表达盒染色体插入位点序列、生化分析、抗生素敏感性/耐药分析； AP-m36.4表达水平； C) 建立 MucoCept-Ab 菌株在雌性恒河猴阴道定殖的能力。目标 2：MucoCept-Ab 方法开发：A) 开发抗体和 ELISA 来检测和测量 AP-m36.4 水平； B) 开发 qPCR 检测来测量 MucoCept-Ab 菌株的水平。目标 3：MucoCept-Ab 配方、工艺开发和制造：A) 开发配方和冻干工艺以生产稳定的 MucoCept-Ab 产品； B) 将 MucoCept-Ab 菌株和技术转移给 GMP 制造商用于生产 MCB 和 WCB，扩大工艺开发规模，并生产 MucoCept-Ab 用于临床前安全性研究。目标 4：MucoCept-Ab 临床前安全性和有效性：A) 评估小鼠阴道给药后 MucoCept-Ab 菌株和纯化的 AP-m36.4 蛋白的急性毒性和全身暴露； B) 评估配制的 MucoCept-Ab 在兔阴道刺激模型中的急性毒性； C) 评估雌性恒河猴阴道给药 MucoCept-Ab 产品后的亚慢性毒性、阴道定植和 AP-m36.4 水平； D) 使用重复的阴道 SHIV 攻击方案评估 MucoCept-Ab 产品在雌性恒河猴中的功效。 如果成功，这种方法可以提供一种安全但廉价的方法，为阴道粘膜提供针对艾滋病毒的被动免疫，并满足对女性控制方法预防性传播病毒感染的迫切需求。

项目成果

Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus.

• DOI: 10.1089/aid.2015.0378
• 发表时间: 2016
• 期刊: AIDS research and human retroviruses
• 影响因子: 1.5
• 作者: Marcobal A;Liu X;Zhang W;Dimitrov AS;Jia L;Lee PP;Fouts TR;Parks TP;Lagenaur LA
• 通讯作者: Lagenaur LA