R01 Estrogen therapy and APOE4 risk in Alzheimer's tested in female EFAD mice

R01 在雌性 EFAD 小鼠中测试雌激素治疗和阿尔茨海默病的 APOE4 风险

• 批准号: 9914419
• 负责人: MARY JO LADU
• 金额: $ 7.08万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9914419
• 关键词: ATP binding cassette transporter 1; Ablation; Affect; Alleles; Alternative Therapies; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease risk; Amyloid; Amyloid beta-Protein; Animal Model; Apolipoprotein E; Apolipoproteins; Attenuated; Behavior; Benchmarking; Biological Availability; Biological Markers; Brain; Clinical; Cognition; Cognitive deficits; Data; Dementia; Development; Dose; Epidemic; Estradiol; Estrogen Receptor alpha; Estrogen Receptor beta; Estrogen Receptors; Estrogen Therapy; Estrogens; Exhibits; Exposure to; FDA approved; Female; Future; Genes; Genotype; Goals; Gynecology; Hot flushes; Human; Impaired cognition; In Vitro; Inflammation; Inflammatory; Link; Measures; Memory; Menopause; Modeling; Mus; Mutation; Neurotoxins; Oral; Outcome; Ovariectomy; Pathology; Pathway interactions; Perimenopause; Pharmacology; Plasma; Postmenopausal Osteoporosis; Postmenopause; Premature Menopause; Premenopause; Presenile Alzheimer Dementia; Prophylactic treatment; Protein Isoforms; Raloxifene; Risk; Role; Safety; Sampling; Selective Estrogen Receptor Modulators; Testing; Therapeutic; Transgenic Mice; Treatment Protocols; Woman; Women' s Health; abeta accumulation; age related; aging brain; apolipoprotein E-4; attenuation; base; cancer chemoprevention; cytokine; dementia risk; design; drug discovery; familial Alzheimer disease; genetic risk factor; improved; in vitro Assay; in vitro Model; in vivo; lifetime risk; male; malignant breast neoplasm; mitochondrial dysfunction; mouse model; nanomolar; novel; pre-clinical; programs; response; sex; virtual; young woman

The gene that controls expression of the apolipoprotein ε4 allele, APOE4, is the greatest genetic risk factor for Alzheimer's disease (AD), increasing risk up to 15-fold compared to APOE3. Although this risk, associated with enhanced amyloid-β (Aβ) accumulation, was discovered over 20 years ago, it has rarely been the focus of therapeutic approaches; even less so, the critical link between female sex and the APOE4-induced risk for dementia and AD. Female (♀) APOE4 carriers have a greater lifetime risk for developing AD, an increased rate of cognitive decline and accelerated accumulation of Aβ compared to male (♂) APOE4 carriers, data consistent with observations in ♀ and ♂ familial AD (FAD) transgenic mice. Estradiol (E2) would appear to be key to this vulnerability: ablation of circulating E2 in pre-menopausal women by oophorectomy causes later cognitive deficits that are reversed by estrogen therapy (ET). The effects of ET after natural menopause remain controversial, particularly with regard to the “critical window” hypothesis stating that ET is beneficial only in the early menopause window. A further concern is the unfavorable safety profile associated with ET resulting from the Women's Health Initiative studies, requiring the development of safer ET alternatives (alt-ET). The goal of this proposal is to determine the ability of ET and alt-ET to counteract the negative interaction of sex with APOE4 in the novel preclinical EFAD mouse model (expressing human APOE +FAD mutations), establishing both the preferred timing and APOE isoform selectivity of safe alt-ET for therapy or prophylaxis of AD. The EFAD mouse was developed to study the interaction between human h-APOE and AD pathology by introducing the h-APOE genotypes into 5xFAD mice: EFAD mice display advanced pathology in females vs. males and E4FAD vs. E3FAD; and preliminary EFAD data suggests that ET after ovariectomy (OVX) protects against cognitive deficits. Alt-ET to be studied are clinical selective estrogen receptor (ER) modulators (SERMs), raloxifene (Ral) and bazedoxifene (Baz); the clinical Baz/E2 combination; and selective estrogen mimics (SEMs) with attenuated gynecological effects. Aim 1: Establish the efficacy of ET (E2) treatment in OVX ♀EFAD mice with respect to APOE genotype, measuring behavior/memory, apoE and Aβ biomarkers, and AD pathology. Aim 2: Establish the “critical window” for ET in EFAD mice with respect to genotype. Aim 3: Test alt-ET in female EFAD mice as transformational AD therapeutics. Dosing will be defined by measuring brain concentrations related to in vitro Kd and EC50. To probe the roles of ER isoforms, mixed glial cultures from APOE-TR mice, will be studied using selective pharmacological ER-probes. In vitro biomarkers (apoE, Aβ, ABCA1, cytokines) will mirror in vivo biomarkers, allowing correlation with in vivo effects of ET and alt-ET, and development of an in vitro assay for future discovery and optimization of Alt-ET for AD.

控制载脂蛋白 ε4 等位基因 APOE4 表达的基因是最大的遗传风险因素 阿尔茨海默病 (AD) 的风险比 APOE3 增加高达 15 倍。尽管这种风险与 β淀粉样蛋白（Aβ）积累增强，20多年前就被发现，但很少成为人们关注的焦点 治疗方法；更不用说，女性性别与 APOE4 引起的风险之间的关键联系 痴呆症和AD。女性 (♀) APOE4 携带者一生中患 AD 的风险更大，发病率更高 与男性 (♂) APOE4 携带者相比，认知能力下降和 Aβ 加速积累，数据一致 在 ♀ 和 ♂ 家族性 AD (FAD) 转基因小鼠中进行观察。雌二醇 (E2) 似乎是其中的关键 脆弱性：绝经前女性通过卵巢切除术消除循环 E2 会导致后来的认知缺陷 雌激素治疗（ET）可以逆转这些症状。自然绝经后 ET 的影响仍存在争议， 特别是关于“关键窗口”假说，该假说指出 ET 仅在早期才有益处 更年期窗口。另一个担忧是与 ET 相关的不利安全状况，这是由于 妇女健康倡议研究，要求开发更安全的 ET 替代品 (alt-ET)。此举的目标 提案的目的是确定 ET 和 alt-ET 抵消性别与 APOE4 的负面相互作用的能力 新型临床前 EFAD 小鼠模型（表达人类 APOE +FAD 突变），建立了 安全 alt-ET 治疗或预防 AD 的首选时机和 APOE 同工型选择性。 EFAD 鼠标 通过引入 h-APOE 来研究人类 h-APOE 与 AD 病理学之间的相互作用 5xFAD 小鼠的基因型：EFAD 小鼠在雌性与雄性之间以及 E4FAD 与雄性之间表现出高级病理学。 E3FAD； EFAD 的初步数据表明，卵巢切除术 (OVX) 后进行 ET 可预防认知缺陷。 待研究的 Alt-ET 包括临床选择性雌激素受体 (ER) 调节剂 (SERM)、雷洛昔芬 (Ral) 和 巴多昔芬（Baz）；临床 Baz/E2 组合；和选择性雌激素模拟物（SEM），具有减毒作用 妇科作用。目标 1：确定 ET (E2) 治疗 OVX ♀EFAD 小鼠的功效 APOE 基因型、测量行为/记忆、apoE 和 Aβ 生物标志物以及 AD 病理学。目标 2：建立 EFAD 小鼠 ET 基因型的“关键窗口”。目标 3：在雌性 EFAD 小鼠中测试 alt-ET 转化性 AD 疗法。剂量将通过测量与体外 Kd 相关的脑浓度来确定 和 EC50。为了探究 ER 亚型的作用，将使用 APOE-TR 小鼠的混合神经胶质培养物进行研究 选择性药理学 ER 探针。体外生物标志物（apoE、Aβ、ABCA1、细胞因子）将在体内得到反映 生物标志物，允许与 ET 和 alt-ET 的体内效应相关，并开发体外测定 Alt-ET 用于 AD 的未来发现和优化。