BASE TITLE: PREVENT PRECLINICAL DRUG DEVELOPMENT PROGRAM: PRECLINICAL EFFICACY AND INTERMEDIATE ENDPOINT BIOMAKERSTASK ORDER TITLE: VACCINES AGAINS

基本标题：预防临床前药物开发计划：临床前疗效和中间终点 BIOMAKERSTASK 订单标题：再次疫苗

• 批准号: 10651935
• 负责人: MARGIE CLAPPER
• 金额: $ 61.42万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2023-12-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10651935
• 关键词: APC gene; Adult; Adverse effects; Advisory Committees; African American; Age; American; Anaerobic Bacteria; Animals; Antibiotics; Antigens; ApcMin/+ mice; Bacillus; Bacteria; Bacteroides fragilis; Binding; Biological; COVID-19; Cancer Etiology; Cell physiology; Cell-Mediated Cytolysis; Chemopreventive Agent; Child; Clinical; Codon Nucleotides; Colitis; Colonoscopy; Colorectal; Colorectal Adenoma; Colorectal Cancer; Colorectal Neoplasms; Colorectal Polyp; Containment; DNA; Data; Dental Plaque; Development; Diagnosis; Disease; Distant; Distant Metastasis; Dysplasia; Encapsulated; Enhancers; Epitopes; Familial Adenomatous Polyposis Syndrome; Family history of; Feces; Fusobacteria; Fusobacterium nucleatum; Genetic; Germ-Line Mutation; Goals; Gram-Negative Bacteria; Helicobacter pylori; Hepatitis B Virus; Hereditary Nonpolyposis Colorectal Neoplasms; Human; Human Papilloma Virus Vaccine; ITIM; Immune; Immune Sera; Immunity; Immunoglobulins; Immunosuppression; Incidence; Individual; Infection; Infection prevention; Infectious Agent; Inflammatory Bowel Diseases; Inherited; Innate Immune Response; Intervention; Lead; Lesion; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of gastrointestinal tract; Measures; Mediating; Membrane; Messenger RNA; Methodology; Microsatellite Instability; Mismatch Repair; Mismatch Repair Deficiency; Modeling; Modification; Morbidity - disease rate; Natural Killer Cells; Neoplasm Metastasis; Non-Insulin-Dependent Diabetes Mellitus; Nonsense Mutation; Obesity; Oncogenic; Operative Surgical Procedures; Oral; Organ; Patients; Phase; Population; Preclinical Drug Development; Prevalence; Preventive; Preventive measure; Preventive service; Preventive vaccine; Primary carcinoma of the liver cells; Program Development; Proteins; Proteomics; Pseudouridine; RNA; RNA vaccine; Race; Recommendation; Recording of previous events; Regimen; Regulatory T-Lymphocyte; Reporter; Resistance; Risk; Risk Factors; Role; Small Intestinal Neoplasm; Survival Rate; Syndrome; System; T-Lymphocyte; Time; Tissues; Tumor Escape; United States; United States National Library of Medicine; Update; Vaccination; Vaccine Antigen; Vaccines; Vesicle; Virulence Factors; Wild Type Mouse; adenoma; aged; base; biomaterial compatibility; cancer risk; cigarette smoking; colorectal cancer risk; colorectal cancer screening; coronavirus disease; design; draining lymph node; effector T cell; efficacy testing; gene repair; glycerophosphodiester phosphodiesterase; immunogenic; immunogenicity; improved; in vivo; infection rate; lipid nanoparticle; malignant breast neoplasm; malignant stomach neoplasm; microbiota; mortality; mouse model; mutation carrier; neoplastic cell; pathogen; preclinical efficacy; premalignant; prevent; prophylactic; receptor; scale up; screening; tumor; tumor growth; tumor microenvironment; tumorigenesis; tumorigenic; vaccine platform; vaccinology

Colorectal cancer (CRC) is one of the leading causes of cancer-related mortality worldwide. In the Unites States, an estimated 151,300 Americans will be diagnosed with CRC and over 52,580 are expected to die from the disease in 2022 alone (https://seer.cancer.gov/statfacts/html/colorect.html). While the five-year survival rate for localized CRC is excellent at 90%, more than half of new CRC cases have the disease spread to regional lymph nodes and/or distant organs at the time of diagnosis. CRC with distant metastasis has a dismal five-year survival rate of 14.7%. These data clearly indicate that CRC-related mortality can be significantly improved if the disease is detected early and proper interventions could be deployed. Because CRC typically originates from precancerous colorectal polyps, routine colonoscopy screening provides an excellent opportunity to detect precursor or early lesions and reduce CRC-related morbidity and mortality. Indeed, the US Preventive Services Task Force recently updated its recommendation on CRC screening, noting the evidence that the screening in average-risk asymptomatic adults aged 50 to 75 years is of substantial benefit. Risk factors for CRC include age (>50), race (African American), obesity, cigarette smoking, type II diabetes, history of inflammatory bowel diseases, family history of colorectal polyps or CRC, and inherited genetic syndromes known to increase the CRC risks, such as familial adenomatous polyposis (FAP) and Lynch syndrome. FAP and Lynch syndrome are caused by germline mutations in the APC gene and DNA mismatch repair genes, respectively. The management of individuals with hereditary gastrointestinal cancer syndromes requires additional measures beyond what is recommended for the average-risk population to minimize the overall risk of cancer-associated morbidity and mortality. Management options for the confirmed mutation carriers include multiple aggressive screening, chemopreventive strategies, and prophylactic surgery. However, these interventions are associated with various degrees of adverse effects. Safer and more effective preventive measures are urgently needed for the individuals with FAP, Lynch syndrome, and other hereditary gastrointestinal cancer syndromes. Fusobacteria are common human oral gram-negative anaerobic microflora isolated from dental plaque and gum diseases, but rarely detectable in the colorectum of healthy individuals. In recent years, the enrichment of a specific pathotype, Fusobacterium nucleatum (Fn), has been demonstrated in the colonic tissues and stools from patients with colorectal adenomas and CRC. Clinical evidence suggests that the prevalence of Fn progressively increases from dysplasia, adenomas to CRC and the higher amount of Fn is significantly associated with CRC with high microsatellite instability, which is caused by DNA MMR deficiencies as seen in Lynch syndrome-associated CRC. The potential association of Fn with CRC tumorigenesis has been examined in a mouse model of FAP. ApcMin/+ mice, which carry a nonsense mutation at codon 850 of the Apc gene, had a significantly accelerated onset and increased multiplicity of both small intestinal and colorectal tumors after oral inoculation of Fn. These data strongly suggested the tumor-promoting role of Fn in ApcMin/+ mice. Interestingly, Fn did not induce colitis in these animals, in contrast to enterotoxigenic Bacteroides fragilis, which has been shown to cause colitis and accelerate tumorigenesis in ApcMin/+ mice. It has also been demonstrated that Fn promotes CRC, breast, and cervical cancer metastasis and can metastasizes with tumor cells. Although how Fn contributes to CRC tumorigenesis has yet to be fully elucidated, emerging evidence points to its virulence factors, such as FadA and Fap2, as potential enhancers of CRC tumorigenesis and progression. The Fap2 has been shown to bind to human T-cell immunoglobulin ITIM domain (TIGIT), an immunoglobulin superfamily receptor known to function as an immune co-inhibitory molecule. Fap2-binding to TIGIT on NK and other T cells protects tumor cells from NK cell-mediated cytotoxicity and blocks effector T cell functions in the tumor microenvironment. TIGIT has also been shown to promote Treg function. Taken together, Fn may exert tumor promoting effects not only by promoting CRC tumor growth, but also by exploiting the immune-suppressive function of TIGIT via its virulence factor protein and contributing to tumor immune evasion mechanisms. Cancers caused by infectious agents are theoretically preventable, if one can prevent the infection, eradicate oncogenic pathogens before tumor development, or suppress the functions of oncogenic molecules. Prophylactic vaccines for human papillomavirus and hepatitis B virus have been associated with significant reductions in infection rates, which are expected to result in the dramatic decrease in the incidence of cervical cancers and hepatocellular carcinoma, respectively. Compared to the prevention of infection, the eradication of oncogenic infectious agents already colonized in the host is in general highly challenging, if not impossible. For example, the well-established eradication regimens for Helicobacter pylori, which increases the risk of noncardia gastric cancers, do not eradicate the bacteria in all cases. Rather, emerging resistance to multiple antibiotics appears to be contributing to the recent decline in the eradication rate. Alternatively, the host immune defense system may be fortified by pathogen-targeted vaccines, if they can boost anti-pathogen immunity and lead to the containment of infection or suppression of pathogen-mediated tumorigenic functions. Fn is a gram-negative anaerobic bacillus, and can be isolated from 60-70% of children aged 5-7 years. As with other gram-negative bacteria, Fn produces outer membrane vesicles (OMVs), which contain much of the biological content of the Fn, but without replicative capacity and some of the soluble proteins found in OMVs may elicit anti-Fn immunity. As part of the PREVENT project to develop Fn OMV based anti-Fn vaccine (https://reporter.nih.gov/project-details/9358850), the proteomic characterization of Fn OMV led to the identification of putative immunogenic components, some of which may be useful as anti-Fn vaccine antigens. Lipid nanoparticle (LNP)-encapsulated mRNA vaccines against COVID-19 revolutionized the implementation of mRNA-based vaccinology. In addition to modifications of mRNA such as with pseudouridine incorporation to reduce innate immune responses and mRNA purification methodologies to remove contaminants, the development of biocompatible LNPs significantly boosted the advancement of COVID and other mRNA-based vaccines. LNP-RNA vaccine platform offers several advantages over conventional protein-based vaccination, including a rapid development, refined adjustment of antigenic epitopes, easier scale-up and timely deployment. The the current study aims to develop and evaluate the immunogenicity of soluble protein-based and LNP-RNA-based Fn vaccines and to determine anti-Fn activity and immunopreventive efficacy of Fn-associated CRC, using mouse models of FAP and Lynch syndrome.

结直肠癌（CRC）是全球与癌症相关死亡率的主要原因之一。在联合国家中，估计将有151,300名美国人被诊断为CRC，仅2022年就会死于该疾病（https://seer.cancer.gov/statfacts/htfacts/html/colectml/colect.html）。虽然局部CRC的五年生存率在90％的情况下非常出色，但在诊断时，有超过一半的新CRC病例扩散到区域淋巴结和/或远处器官。远处转移的CRC的五年生存率为14.7％。这些数据清楚地表明，与CRC相关的死亡率可以显着提高，如果疾病早期发现并可以部署适当的干预措施。由于CRC通常起源于癌前大息息肉，因此常规结肠镜检查为检测前体或早期病变并降低与CRC相关的发病率和死亡率提供了绝佳的机会。确实，美国预防服务工作队最近更新了有关CRC筛查的建议，并指出证据表明，在50至75岁的平均无症状的无症状成年人中进行筛查具有很大的好处。 CRC的危险因素包括年龄（> 50），种族（非洲裔美国人），肥胖，吸烟，II型糖尿病，炎症性肠病史，大肠息肉或CRC的家族史以及已知的遗传综合征，以增加CRC风险，例如CRC风险，例如家族性腺瘤性息肉病（FAP）和乳液综合征。 FAP和Lynch综合征分别是由APC基因和DNA不匹配修复基因中的种系突变引起的。遗传性胃肠道综合症患者的管理需要额外的措施，而不是为平均风险人群推荐的措施，以最大程度地减少与癌症相关的发病率和死亡率的总体风险。确认的突变载体的管理选择包括多次侵略性筛查，化学预防策略和预防性手术。但是，这些干预措施与不同程度的不良反应有关。对于患有FAP，Lynch综合征和其他遗传性胃肠道癌综合征的患者，迫切需要采取更安全，更有效的预防措施。 葡萄球菌是常见的人类口服革兰氏阴性厌氧症，从牙菌斑和牙龈疾病中分离出来，但在健康个体的结肠菌性中很少检测到。近年来，在结肠组织中已证明了特定病原体核细菌（FN）的富集和结直肠腺瘤和CRC患者的粪便。临床证据表明，FN的患病率逐渐从发育不良，腺瘤到CRC逐渐增加，而较高的FN与CRC的CRC显着相关。在FAP的小鼠模型中，已经检查了FN与CRC肿瘤发生的潜在关联。 APCMIN/+小鼠在APC基因的密码子850处携带无义突变，在口服FN口服后，小肠和结直肠肿瘤的多样性显着加速，并增加了多重性。这些数据强烈建议FN在APCMIN/+小鼠中促进肿瘤的作用。有趣的是，与肠毒素菌菌素Fragilis相比，FN并未诱导这些动物的结肠炎，而肠毒性杆菌叶片已被证明会引起结肠炎并加速apcmin/+小鼠的肿瘤发生。还证明FN促进CRC，乳腺癌和宫颈癌转移，并可以与肿瘤细胞转移。 尽管FN如何促进CRC肿瘤发生尚未完全阐明，但新兴的证据表明其毒力因子（例如FADA和FAP2）是CRC肿瘤发生和进展的潜在增强子。 FAP2已显示与人类T细胞免疫球蛋白ITIM结构域（Tigit）结合，这是一种免疫球蛋白超家族受体，被称为免疫抑制分子。 FAP2结合NK和其他T细胞上的TigIT可保护肿瘤细胞免受NK细胞介导的细胞毒性的影响，并阻止肿瘤微环境中的T细胞功能。 Tigit还显示出可以促进Treg功能。综上所述，FN不仅可以通过促进CRC肿瘤的生长来促进肿瘤促进作用，还可以通过其通过其毒力因子蛋白来利用TigIT的免疫抑制功能，并有助于肿瘤免疫逃避机制。 由传染剂引起的癌症在理论上是可以预防的，如果可以预防感染，请在肿瘤发育前消除致癌病原体，或抑制致癌分子的功能。人乳头瘤病毒和丙型肝炎病毒的预防性疫苗与感染率的显着降低有关，这有望导致宫颈癌和肝细胞癌的发生率急剧下降。与预防感染相比，根除在宿主中已经殖民的致癌感染剂通常是高度挑战，即使不是不可能。例如，在所有情况下，都不会消除细菌的幽门螺杆菌的良好成熟的根除方案，从而增加了胃癌的胃癌风险。相反，对多种抗生素的新兴耐药性似乎导致了最近的根除率下降。 或者，如果宿主免疫防御系统可以通过靶向病原体的疫苗来加强，如果它们可以增强抗病原体免疫并导致感染或抑制病原体介导的肿瘤功能。 FN是一种革兰氏阴性的厌氧菌，可以从60-70％的5-7岁儿童中分离出来。与其他革兰氏阴性细菌一样，FN产生外膜囊泡（OMV），其中包含FN的许多生物含量，但没有复制能力，OMV中发现的某些可溶性蛋白可能会引起抗FN免疫。作为预防项目的一部分，开发基于FN OMV的抗FN疫苗（https://reporter.nih.gov/project-details/9358850），FN OMV的蛋白质组学表征导致鉴定出了假定的免疫原性成分，其中一些可能是有用的，可能是抗抗抗原抗生素。 脂质纳米颗粒（LNP）封装的mRNA疫苗针对COVID-19彻底改变了基于mRNA的疫苗学的实施。除了对mRNA的修改（例如假氨酸掺入）以减少先天免疫反应和mRNA纯化方法以去除污染物，而生物相容性的LNP的发展显着增强了共vid和其他mRNA基疫苗的发展。 LNP-RNA疫苗平台比常规蛋白质疫苗接种提供了多个优点，包括快速开发，精致的抗原表位调整，更容易扩展和及时的部署。当前的研究旨在使用FAP和Lynch综合征的小鼠模型来开发和评估基于可溶性蛋白质和LNP-RNA的FN疫苗的免疫原性，并确定与FN相关CRC的抗FN活性和免疫预防疗效。