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: 8658658
• 负责人: Laurel A. Lagenaur
• 金额: $ 133.55万
• 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 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8658658
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Acute; Address; Antibiotic susceptibility; Antibodies; Ascaridil; 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; 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; 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; public health relevance; 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。从生物学上讲，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，包括詹森乳杆菌，有助于保护这些表面，并通过入侵病原体来阻止定植和感染。流行病学研究表明，阴道乳酸菌的丧失与异性传播艾滋病毒-1和其他性传播感染的风险增加有关。Osel公司利用这些天然保护剂的特性，设计了一种天然的人阴道乳杆菌L.jensenii 1153，以表达针对HIV的有效和广谱中和单域抗体，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L.jensenii的HIV-1进入抑制剂MCV-N，它将SIV病毒在猕猴中的传播减少了63%(Lagenaur等人，Mucosal Invol，(2011年7月6日))。第二阶段的研究将推进MucoCept-Ab的临床前开发。这些研究将集中于开发安全、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、临床前安全性测试和非人灵长类(NHP)病毒挑战模型的有效性测试。这一阶段的目标是开发一种能够在局部传递抗体的乳杆菌菌株，以防止艾滋病毒在阴道或直肠粘膜上的感染。这种菌株将被配制成一种成本效益高的产品，能够提供持续的保护，并由女性控制。目的1：MucoCept-AbAPI的优化和鉴定：a)完整的MucoCept-Ab株优化：最大化启动子强度，优化简氏乳杆菌密码子，去除表位标签，并测试两个基因组整合位点；b)MucoCept-Ab株的完整微生物学特性；遗传稳定性，表达盒序列染色体插入位置，生化图谱，抗生素敏感性/耐药性图谱；AP-m36.4表达水平；c)建立MucoCept-Ab株经阴道定植雌性恒河猴的能力。目的2：MucoCept-Ab方法的发展：A)建立抗体和ELISA检测AP-m36.4水平；B)建立定量PCR方法检测MucoCept-Ab株的水平。目的3：MucoCept-Ab的处方、工艺开发和制造：A)开发处方和冻干工艺，以生产稳定的MucoCept-Ab产品；B)将MucoCept-Ab的菌株和技术转移到GMP制造商，用于生产MCB和WCB，扩大工艺开发，并生产用于临床前安全性研究的MucoCept-Ab。目的4：MucoCept-Ab的临床前安全性和有效性：A)评估MucoCept-Ab株和纯化的AP-m36.4蛋白经阴道给药后的急性毒性和全身暴露；B)在兔阴道刺激模型中评估配制的MucoCept-Ab的急性毒性；C)评估MucoCept-Ab产品经阴道给药后对雌性恒河猴的亚慢性毒性、阴道定植和AP-m36.4水平；D)采用重复阴道SHIV激发方案评价MucoCept-Ab产品对雌性恒河猴的疗效。如果成功，这种方法可以提供一种安全而廉价的手段，向阴道粘膜提供对艾滋病毒的被动免疫，并满足对女性控制方法的迫切需要，以防止性传播病毒感染。