Second-generation Gene Transfer Vector-based Anti-cocaine Vaccines

第二代基因转移载体抗可卡因疫苗

• 批准号: 8439372
• 负责人: RONALD G CRYSTAL
• 金额: $ 81.19万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8439372
• 关键词: Active immunity; Addictive Behavior; Adenovirus Protein; Adenoviruses; Adjuvant; Affinity; Animals; Antibodies; Antigen-Presenting Cells; Antigens; Behavior; Blood; Blood Circulation; Brain; Breathing; Capsid; Capsid Proteins; Cocaine; Cocaine Dependence; Code; Coupled; Data; Dependovirus; Dose; Exhibits; Extinction (Psychology); Fiber; Gene Transfer; Generations; Human; Hyperactive behavior; Immune system; Immunity; Intravenous; Knowledge; Link; Lysine; Medical; Modeling; Monoclonal Antibodies; Mus; Nose; Passive Immunity; Phenotype; Proteins; Rattus; Rewards; Rodent; Self Administration; Serotyping; Site; Technology; Testing; Vaccinated; Vaccines; adeno-associated viral vector; analog; anti-IgG; base; cocaine use; experience; gene transfer vector; immunogenic; immunogenicity; improved; nonhuman primate; novel; prevent; public health relevance; receptor; small molecule; social; success; vaccine development

DESCRIPTION (provided by applicant): One approach to treating cocaine addiction is to vaccinate against cocaine, preventing it from reaching the brain. The challenge is that cocaine, like most small molecules, is a poor immunogen. Based on our experience with adenovirus (Ad) gene transfer vectors in animals and humans, and the recognition that Ad is a potent adjuvant that activates the immune system, we hypothesized that cocaine analogs linked to Ad capsid proteins would elicit high level, high affinity cocaine specific antibodies sufficient to trat cocaine addiction. Our 1st approach was to link the cocaine analogs GNC or GNE to a disrupted E1-E3- serotype 5 Ad (dAd5). The dAd5GNC vaccine elicited high affinity (Kd 45 nM) IgG anti-cocaine titers in mice, and vaccinated mice no longer responded with hyperactive behavior following repetitive intravenous doses of 50 ?g cocaine. dAd5GNE evoked higher titers than dAd5GNC, and dAd5GNE-vaccinated rats exhibited suppressed cocaine reward, no extinction "burst" of activity seen in non-vaccinated rats, and did not reinstate cocaine seeking following a cocaine prime. The focus of the 3 aims of this proposal is to develop 2nd generation anti-cocaine vaccines that build on the success of dAd5GNC and dAd5GNE, enabling lower doses to generate higher titer, higher affinity anti-cocaine antibodies that abrogate the activity and self-administration phenotypes associated with cocaine administration. Aim 1. The capsid hexon and fiber are the most immunogenic Ad proteins. We hypothesize that the cocaine analog GNE coupled directly to purified hexon and/or fiber will generate a more potent vaccine than GNE coupled to the entire disrupted Ad. Further, the hexon and fiber sequences will be genetically modified to include additional lysine residues, increasing covalent attachment sites for GNE. Aim 2. Immunity against Ad capsid proteins limit efficacy of Ad vaccines. Since different Ad serotypes stimulate immunity differently, we hypothesize that repeated alternating administration of GNE coupled to disrupted Ad5 and sAd36 (a highly immunogenic nonhuman primate Ad) or alternating administration of GNE coupled to purified hexons/fibers of each serotype, will evoke higher titers than with a single serotype. Aim 3. We have also developed a "persistent passive anti-cocaine immunity" strategy using an adeno-associated virus serotype rh.10 coding for an anti-cocaine monoclonal antibody (AAVrh.10antiCoc) that generates persistent anti-cocaine antibodies sufficient to suppress cocaine induced hyperactivity in mice. We hypothesize that AAVrh.10antiCoc either alone, or together with the best active Ad vaccines from aims 1 and 2, will provide highly effective anti-cocaine protection to the CNS. An AAV coding for anti-cocaine will also be administered to the nose to determine if high titer anti-cocaine antibodies at the site of entry can aid as a 1st anti-cocaine defense.

描述（由申请人提供）：治疗可卡因成瘾的一种方法是接种可卡因疫苗，防止其到达大脑。挑战在于可卡因，像大多数小分子一样，是一种弱免疫原。 基于我们在动物和人类中使用腺病毒（Ad）基因转移载体的经验，以及认识到Ad是激活免疫系统的有效佐剂，我们假设与Ad衣壳蛋白连接的可卡因类似物将引发足以治疗可卡因成瘾的高水平、高亲和力可卡因特异性抗体。我们的第一种方法是将可卡因类似物GNC或GNE与破坏的E1-E3-血清型5 Ad（dAd 5）连接。dAd 5GNC疫苗引起高亲和力（Kd 45 nM）IgG抗可卡因滴度在小鼠中，和接种疫苗的小鼠不再与多动行为后，重复静脉注射剂量为50？g可卡因。dAd 5GNE诱发比dAd 5GNC更高的滴度，并且dAd 5GNE接种的大鼠表现出抑制的可卡因奖赏，在未接种的大鼠中没有观察到活性的消退“爆发”，并且在可卡因引发后没有恢复可卡因寻求。该提案的3个目标的重点是开发第二代抗可卡因疫苗，其建立在dAd 5GNC和dAd 5GNE的成功基础上，使得较低剂量能够产生较高滴度、较高亲和力的抗可卡因抗体，其消除与可卡因施用相关的活性和自我施用表型。目标1.衣壳六邻体和纤维是最具免疫原性的Ad蛋白。我们假设直接偶联至纯化的六邻体和/或纤维的可卡因类似物GNE将产生比偶联至整个破坏的Ad的GNE更有效的疫苗。此外，六邻体和尾丝序列将被遗传修饰以包括额外的赖氨酸残基，增加GNE的共价连接位点。目标2.针对Ad衣壳蛋白的免疫限制了Ad疫苗的功效。由于不同的Ad血清型不同地刺激免疫，我们假设重复交替施用与破坏的Ad 5和sAd 36（高度免疫原性的非人灵长类Ad）偶联的GNE或交替施用与每种血清型的纯化的六邻体/纤维偶联的GNE，将引起比单一血清型更高的滴度。目标3.我们还开发了一种“持续被动抗可卡因免疫”策略，其使用编码抗可卡因单克隆抗体（AAVrh.10antiCoc）的腺相关病毒血清型rh.10，其产生足以抑制可卡因诱导的小鼠多动症的持续抗可卡因抗体。我们假设AAVrh.10antiCoc单独或与来自目标1和2的最佳活性Ad疫苗一起将为CNS提供高度有效的抗可卡因保护。还将编码抗可卡因的AAV施用至鼻以确定进入位点处的高滴度抗可卡因抗体是否可以作为第一抗可卡因防御来帮助。