Cryopreservation of Anopheles stephensi embryos

史氏按蚊胚胎的冷冻保存

• 批准号: 8117546
• 负责人: Peter F. Billingsley
• 金额: $ 30万
• 依托单位: SANARIA, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8117546
• 关键词: Adult; Africa; African; Anopheles Genus; Attenuated; Biology; Biotechnology; Bite; Blood; Cessation of life; Characteristics; Child; Clinical; Clinical Trials; Cryopreservation; Culicidae; Development; Disclosure; Embryo; Ensure; Erythrocytes; Fertility; Genetic; Genetic Drift; Goals; Health; Human Volunteers; Infant; Infection; Institutes; Lead; Licensure; Life Cycle Stages; Liquid substance; Longevity; Maintenance; Malaria; Malaria Vaccines; Marketing; Maryland; Measures; Methods; Military Personnel; Nitrogen; Outcome; Parasites; Phase; Phase I Clinical Trials; Pilot Projects; Plasmodium falciparum; Predisposition; Preparation; Procedures; Process; Production; Protocols documentation; Quality Control; Safety; Salivary Glands; Secure; Security; Seeds; Site; Small Business Innovation Research Grant; Source; Sporozoite vaccine; Sporozoites; Staging; Sterility; Technology; Testing; Transgenic Organisms; Universities; Vaccines; cell bank; clinical lot; cost; disease transmission; egg; hatching; immunogenicity; manufacturing process; mortality; novel; phase 2 study; phase 3 study; prevent; programs; protective efficacy; scale up; success; tool; transmission process; vapor; vector; young adult

DESCRIPTION (provided by applicant): Malaria is responsible for several hundred million cases and over one million deaths per year. A highly effective vaccine that prevents blood stage infection and thereby prevents all disease and transmission would be an ideal tool for eliminating Plasmodium falciparum (Pf), the cause of 99% of malaria mortality. Attenuated Pf sporozoites (PfSPZ) administered by the bite of infected mosquitoes, lead to protection of >90% of human volunteers against experimental Pf challenge that lasts at least 10 months. Sanaria's goal is to develop and commercialize an attenuated PfSPZ vaccine that prevents Pf blood stage infection in >90% of recipients - a vaccine with these characteristics could be used to eliminate Pf from the world. This vaccine has the potential for >$1 billion annual revenues. Sanaria has successfully established robust, reproducible, and consistent manufacture and release of clinical lots of its PfSPZ Vaccine, received FDA approval (IND) for clinical trials, and initiated a Phase 1 clinical trial to assess safety, immunogenicity, and protective efficacy of the PfSPZ Vaccine in May 2009. Additional proof of concept studies in African adults, young children, and infants are planned and will be followed by Phase 2 studies, and then pivotal Phase 3 studies to support licensure. Sanaria's manufacturing process utilizes a master cell bank of cryopreserved P. falciparum-infected erythrocytes as a source of parasites, and Anopheles stephensi mosquitoes to raise the PfSPZ. However, because there is no method for cryopreserving any stage of the Anopheles mosquito life cycle, the mosquitoes used for producing the PfSPZ Vaccine come from a mosquito colony that is continuously maintained. Cryopreservation of the eggs of Anopheles spp. would enable Sanaria to cryobank different strains of An. stephensi, reduce costs associated with strain maintenance, produce a uniform seed stock and provide security against strain loss or genetic drift. Cryobanking of transgenic An. stephensi and other Anopheles spp. would also provide direct benefit to those aiming to use genetically altered or sterile mosquitoes for transmission control. In pilot studies, we have shown that An. stephensi can be cryopreserved in the vapor phase of liquid nitrogen using a novel cryoprotectant additive (CPA) mixture, with successful hatching and larval development following thawing. An invention disclosure describing the details that led to the production of the first successful Anopheles egg cryopreservation has been filed. The hatch rate to date has been 10% compared to non-cryopreserved controls from the same egg batch which is clear indication of proof of concept. This proposal will define and optimize the successful cryopreservation of An. stephensi by a pragmatic and systematic testing of embryos at different stages of development and of cryopreservation methods and additives. Successful outcome will be measured as the development from cryopreserved eggs of An. stephensi that retain full susceptibility to infection with PfSPZ in the salivary glands. Other strains of An. stephensi held by Sanaria, including transgenic strains, will be similarly cryopreserved. PUBLIC HEALTH RELEVANCE: Malaria causes 500 million clinical cases and 1-3 million deaths annually, is responsible for >1% loss of GDP in Africa annually and is a serious concern for travelers and military personnel. Sanaria's goal is to develop and commercialize a >90% protective malaria vaccine for three primary markets with a potential for >$1 billion annual revenues. Success in this project will represent a major breakthrough in the fields of vector and malaria biology, will ensure the ability to conserve the genetic stock of the mosquitoes used by Sanaria's in its vaccine manufacturing process, will move Sanaria towards ensuring consistency of mosquito colonies across different manufacturing sites and will provide a secure cryobank of material from which mosquito production can be initiated periodically and which can be followed in a well-defined stability program.

描述（由申请人提供）：疟疾造成数亿例病例和每年超过100万人死亡。一种高效的疫苗，可防止血液阶段感染，从而防止所有疾病和传播是消除恶性疟原虫（PF）的理想工具，这是疟疾死亡率99％的原因。被感染蚊子咬伤的减毒PF孢子虫（PFSPZ）导致> 90％的人类志愿者免受至少10个月的实验PF挑战的保护。 Sanaria的目标是开发和商业化降低的PFSPZ疫苗，该疫苗可防止> 90％的受体中PF血液阶段感染 - 具有这些特征的疫苗可用于消除世界上的PF。这种疫苗的年收入有可能获得10亿美元。 Sanaria已成功确立了其PFSPZ疫苗的临床大量临床和释放的强大，可再现和一致的制造，并获得了FDA批准（IND）进行临床试验，并启动了一项1期临床试验，以评估2009年5月的PFSPZ疫苗和其他概念的PFSPZ疫苗，以评估2009年5月的PFSPZ疫苗的效力。研究，然后是关键第三阶段研究以支持许可。 Sanaria的制造过程利用了一个冷冻保存的恶性疟原虫感染的红细胞作为寄生虫来源，而Anopheles Stephensi蚊子来升高PFSPZ。但是，由于没有方法可以冷冻水平的蚊子生命周期的任何阶段，因此用于生产PFSPZ疫苗的蚊子来自不断维持的蚊子菌落。肛门属卵的冷冻保存。将使Sanaria能够冰冻库的不同菌株。斯蒂芬西（Stephensi），降低与应变相关的成本，产生均匀的种子库存，并为应变损失或遗传漂移提供安全性。转基因an的冷冻群。 Stephensi和其他按蚊属。还将为那些旨在使用遗传改变或无菌蚊子进行传输控制的人提供直接利益。在试点研究中，我们已经证明了这一点。史蒂芬西（Stephensi）可以使用新型的冷冻保护剂添加剂（CPA）混合物在液氮的蒸气阶段进行冷冻保存，并在解冻后成功进行孵化和幼虫发育。已经提出了一项发明披露，描述了导致第一个成功的卵卵冷冻保存的细节。迄今为止，迄今为止的舱口速率已为10％，与来自同一鸡蛋批处理的非晶体对照相比，这清楚地表明了概念证明。该建议将定义和优化AN的成功冷冻保存。 Stephensi通过在开发的不同阶段和冷冻保存方法和添加剂的不同阶段对胚胎进行务实的系统测试。成功的结果将被衡量为An的冷冻保存卵的发育。 Stephensi保留了唾液腺中PFSPZ感染的完全敏感性。其他菌株。由Sanaria持有的Stephensi（包括转基因菌株）将同样冷冻保存。 公共卫生相关性：疟疾每年造成5亿次临床病例和1-3万人死亡，每年造成> 1％的GDP损失，这是旅行者和军事人员的严重关注。萨纳里亚（Sanaria）的目标是开发和商业化> 90％的防护性疟疾疫苗，以便三个主要市场的年收入> 10亿美元。该项目的成功将代表媒介和疟疾生物学领域的重大突破，将确保能够保存Sanaria在其疫苗制造过程中使用的蚊子的遗传库存，将使Sanaria迁移Sanaria朝着Sanaria的一致性，以确保在不同制造地点的摩西群落和摩西群岛的生产范围内的生产能力，并能够逐步提供摩西群的生产。 程序。