Development of gene transfer approach for prevention of vaginal HIV transmission

开发预防艾滋病毒阴道传播的基因转移方法

• 批准号: 7496209
• 负责人: Phuong Thi Nguyen-Sarkis
• 金额: $ 4.48万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7496209
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Antibodies; Antiviral Agents; Blocking Antibodies; Cell Line; Cell-Matrix Junction; Cells; Cervical; Cervix Uteri; Chimeric Proteins; Clinical Trials; Dependovirus; Development; Epithelial; Epithelial Cells; Fellowship; Female; Foundations; Gene Transfer; Genital system; Goals; HIV; HIV-1; Hematopoietic stem cells; Human; Humoral Immunities; Immune; Immunity; In Vitro; Infection; Inflammation; Libraries; Mediating; Menstrual cycle; Mus; Peptides; Phage Display; Prevention; Prevention approach; Proteins; Safety; Serotyping; Solid; Surface; System; Target Populations; Testing; Tissue Model; Tissues; Toxic effect; Vaccine Research; Vaccines; Vagina; Viral; Virion; Virus; Woman; base; fibroglycan; gene therapy; genetic vaccine; human tissue; immune function; in vivo; macrophage; member; monocyte; mouse model; neutralizing antibody; nonhuman primate; novel; novel strategies; prophylactic; reconstitution; research study; success; syndecan; transcytosis; transmission process

DESCRIPTION (provided by applicant): More than half of new HIV-1 infections are acquired by women through intravaginal exposure. The cervico-vaginal epithelial cells lining the mucosal surfaces of the female lower genital track provide an initial defense against HIV-1 infection, but protection is incomplete. Transport of HIV-1 across this barrier is critical for HIV-1 colonization and virus dissemination. The long-term goal is to enhance anti-HIV-1 humoral immunity at the mucosal surface by local expression of anti-HIV-1 neutralizing antibodies to block epithelial cell attachment and virus entry. The safety profile, low immunogenecity and rapid advancement of other AAV based gene therapies into human clinical trials makes it a feasible approach to HIV-1 prevention. During my fellowship, I will investigate whether stable adeno-associated virus (AAV)-neutralizing human antibody gene transfer to endocervical, ectocervical and vaginal epithelial cells can provide durable protection against HIV-1. Specifically, I will 1) Determine the AAV serotype (1-9) that provides optimal gene transfer to these cells without toxicity; 2) Isolate new neutralizing human anti-syndecan 1 and anti-syndecan 2 antibodies from a 27 billion member phage display library and test their abilities to block attachment of HIV-1 to human primary cervico-vaginal epithelial cells; and construct a Virus Inhibitory Peptide (VIRIP)-Fc fusion protein and test its ability to inhibit infection by blocking viral fusion. The anti-syndecan antibodies and VIRIP-Fc proteins will be tested for inhibition of virus internalization and transcytosis across the cervico- vaginal epithelial cells and infectivity of the transcytosed virus particles in vitro. 3) Determine if stable AAV- mediated gene transfer can be achieved in the lower genital track of mice with durable intravaginal secretion of neutralizing titers of human anti-syndecan antibodies and VIRIP-Fc fusion proteins. Finally, intravaginal AAV-anti-syndecan and AAV-VIRIP-Fc gene transfer studies followed by intravaginal HIV-1 challenge in a human hematopoietic stem cell-engrafted mice model will provide an experimental test of this novel hypothesis. The success of this novel approach could be advanced to non-human primate studies, and support prophylactic vaccine research with the potential to advance AIDS prophylactic vaccine strategies. PULBIC HEALTH RELEVANCE: HIV-1 infections are acquired most often through sexual contact and more than half of new infections are acquired by women through intravaginal HIV exposure. I propose to develop a genetic vaccine that when delivered to the mucosal surface of the cervix and vagina to allow the lining cells to stably produce human antibodies that block HIV-1 attachment and infection. A protective genetic vaccine delivered to the female lower genital track could dramatically slow the spread of HIV/AIDS.

描述（由申请人提供）：超过一半的新的HIV-1感染是由妇女通过阴道内接触获得的。内衬于女性下生殖道粘膜表面的宫颈阴道上皮细胞提供了对HIV-1感染的初始防御，但保护是不完全的。HIV-1跨越这一屏障的运输对于HIV-1定殖和病毒传播至关重要。长期目标是通过局部表达抗HIV-1中和抗体来阻断上皮细胞附着和病毒进入，从而增强粘膜表面的抗HIV-1体液免疫。其他基于AAV的基因疗法的安全性、低免疫原性和进入人类临床试验的快速进展使其成为预防HIV-1的可行方法。在我的研究期间，我将研究是否稳定的腺相关病毒（AAV）中和人抗体基因转移到宫颈内，宫颈外和阴道上皮细胞可以提供持久的保护，防止艾滋病毒-1。2）从270亿个成员的噬菌体展示文库中分离新的中和性人抗多配体蛋白聚糖1和抗多配体蛋白聚糖2抗体，并测试它们阻断HIV-1与人原代宫颈阴道上皮细胞附着的能力;构建病毒抑制肽（VIRIP）-Fc融合蛋白，并测试其通过阻断病毒融合来抑制感染的能力。将检测抗多配体蛋白聚糖抗体和VIRIP-Fc蛋白对病毒内化和跨宫颈阴道上皮细胞胞吞的抑制作用以及体外胞吞病毒颗粒的感染性。3)确定是否可以在阴道内持久分泌中和滴度的人抗多配体蛋白聚糖抗体和VIRIP-Fc融合蛋白的小鼠下生殖道中实现稳定的AAV介导的基因转移。最后，阴道内AAV-抗-syndecan和AAV-VIRIP-Fc基因转移研究，然后在人造血干细胞移植小鼠模型中进行阴道内HIV-1攻击，将为这种新假设提供实验检验。这种新方法的成功可以推进到非人类灵长类动物研究，并支持预防性疫苗研究，有可能推进艾滋病预防性疫苗策略。 牙髓健康相关性：艾滋病毒-1感染最常通过性接触获得，半数以上的新感染是妇女通过阴道内艾滋病毒感染获得的。我建议开发一种基因疫苗，当被递送到子宫颈和阴道的粘膜表面时，允许衬里细胞稳定地产生阻断HIV-1附着和感染的人类抗体。将保护性基因疫苗注射到女性下生殖道，可以大大减缓艾滋病毒/艾滋病的传播。