A Therapeutic Role for Apolipoprotein-E in the Germ Theory of Alzheimer's Dementia

载脂蛋白-E 在阿尔茨海默氏痴呆病菌理论中的治疗作用

• 批准号: 10601779
• 负责人: Michael P. Vitek
• 金额: $ 50万
• 依托单位: REGENNOVA, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10601779
• 关键词: Adult; Affect; African American population; Age; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; American; Amyloid; Amyloid beta-Protein; Animal Model; Anti-Bacterial Agents; Anti-Inflammatory Agents; Apolipoprotein E; Arginine; Bacteria; Bacterial DNA; Bacterial Infections; Binding; Blood; Brain; Brain Pathology; Caspase; Cell Surface Proteins; Cell surface; Clinic; Communicable Diseases; DNA; Data; Deposition; Disease; Dose; Encephalitis; Germ; Gingivitis; Goals; Gram-Negative Bacteria; Grant; Growth; In Vitro; Infection; Inflammation; Inflammatory; Invaded; Irrigation; Learning; Ligands; Link; Lipopolysaccharides; Longevity; Lysine; Measures; Memory; Minimum Inhibitory Concentration measurement; Modeling; Mus; Oral; Oral Administration; Oral cavity; Outcome; Pathogenicity; Pathology; Patients; Peptides; Performance; Periodontal Diseases; Peripheral; Phase; Porphyromonas gingivalis; Proteins; Qi; Reporting; Ribosomal RNA; Risk; Risk Reduction; Role; Route; Safety; Sepsis; Streptococcus cristatus; Symptoms; Testing; Therapeutic; Tissues; Tooth Loss; Toxicology; United States; Wild Type Mouse; aerobic respiration control protein; blood-brain barrier crossing; cecal ligation puncture; clinically relevant; commensal bacteria; comparison control; cytokine; dementia risk; design; efficacy evaluation; experimental study; gingipain; human old age (65+); improved; inhibitor; mimetics; mouse model; mutant; neuron loss; novel; novel strategies; oral infection; pathogenic bacteria; peptidomimetics; preference; tau-1; theories; virulence gene

Abstract: A Therapeutic Role for Apolipoprotein-E in the Germ Theory of AD P. gingivalis is the keystone bacteria of periodontal disease, the 6th most common infectious disease worldwide. Half of all Americans over the age of 30 and 70% of those over age 65 suffer some degree of P. gingivalis infection associated with gum disease. In a study of 6800 patients with gum disease that were followed for up to 26 years, those that developed gingivitis and then periodontal disease were linked to a significantly increased risk of dementia including Alzheimer's dementia. Tooth loss, a frequent outcome of periodontal disease, was also associated with an increased the risk of dementia. P. gingivalis products including lipopolysaccharide, cysteine proteases known as gingipains, and 16S rRNA from P. gingivalis have all been found in the brains of Alzheimer's patients. Since P. gingivalis is found in the mouth, the blood and is known to invade non-oral tissues including the brain, then strategies to reduce P. gingivalis numbers and activities including reducing brain inflammation, are beginning to show positive effects on outcomes associated with AD in animal models and in the clinic. We recently discovered that small mimetics of Apolipoprotein-E (ApoE-mimetics) inhibited the growth and killed P. gingivalis bacteria. Other groups have reported similar anti-bacterial activities of ApoE-mimetics including our ApoE-mimetics. These anti-bacterial activities appear to correlate with anti-inflammatory activities of ApoE-mimetics reported by us and other groups. We reported that these ApoE-mimetics extended lifespan in mice subjected to whole body sepsis using a caecal ligation and puncture model. We also reported that these ApoE-mimetics cross the blood brain barrier where they reduce brain inflammation in multiple models of Alzheimer's disease. In this grant, we propose to preferentially kill P. gingivalis by a unique strategy. This strategy involves a targeting ligand that specifically binds to a unique cell surface protein on P. gingivalis that, when conjugated with our anti-bacterial ApoE-mimetics, provides a “targeted” anti-bacterial agent. We refer to the “P. gingivalis targeting motif” as a “gingi-tif” and the conjugation with an ApoE-mimetic with as an “ApoE-gingitif”. As proof- of-principle, we now show that the ApoE-gingitif known as RGN2002 retains bacterial killing activity and preferentially targets P. gingivalis killing over killing of commensal bacteria (Figure 9 and Table 2). We will synthesize additional ApoE-gingitif and gingitif-ApoE conjugates and use them to measure the minimum inhibitory concentrations (MICs) against P. gingivalis. We will also use these ApoE-gingitif and gingitif-ApoE conjugates to measure MICs against a non-pathogenic commensal bacteria also found in the mouth. The ratio of the MICs for killing pathogenic P. gingivalis versus non-pathogenic commensal bacteria will provide a quantitation of the preferential killing of P. gingivalis, which we call a “Preference Ratio.” (Table 2). Once our in vitro screen has selected an appropriate ApoE-gingitif or gingitif-ApoE conjugate, then we will test these conjugates in a whole mouse model of P. gingivalis infection. Since oral delivery of P. gingivalis resulted in increased amyloid beta peptide 1-42, increased phosphorylated-tau and increased cytokines, which are all found in Alzheimer's patient's brains, we will measure these products in the brains of P. gingivalis infected mice with and without ApoE-gingitif or gingitif-ApoE conjugate treatment. These results will permit us to determine the efficacy of this targeted “ApoE-gingitif-strategy” to reduce brain inflammation and pathology associated with AD and P. gingivalis infections.

摘要：载脂蛋白 E 在 AD 生殖理论中的治疗作用 牙龈卟啉单胞菌是牙周病的关键细菌，牙周病是第六大常见传染病 全球 30 岁以上的美国人中有一半和 65 岁以上的美国人中有 70% 患有某种程度的疟原虫病。 一项针对 6800 名牙龈疾病患者的研究显示，牙龈炎与牙龈疾病相关。 在长达 26 年的跟踪调查中，那些出现牙龈炎和牙周病的人与 痴呆症的风险显着增加，包括阿尔茨海默氏症痴呆，这是痴呆症的常见后果。 牙周病也与痴呆症风险增加有关。 包括脂多糖、被称为牙龈蛋白酶的半胱氨酸蛋白酶和来自牙龈卟啉单胞菌的 16S rRNA 由于牙龈卟啉单胞菌存在于口腔、血液中，因此所有这些都在阿尔茨海默病患者的大脑中发现。 已知会侵入包括大脑在内的非口腔组织，然后采取策略减少牙龈卟啉单胞菌数量和 包括减少脑部炎症在内的活动开始对相关结果显示出积极影响 AD 在动物模型和临床中的应用。 我们最近发现载脂蛋白-E 的小模拟物 (ApoE-mimetics) 抑制生长 并杀死牙龈卟啉单胞菌细菌其他研究小组也报道了 ApoE 模拟物的类似抗菌活性。 包括我们的 ApoE 模拟物，这些抗菌活性似乎与抗炎相关。 我们和其他小组报告的 ApoE 模拟物的活性 我们报告说，这些 ApoE 模拟物延伸了。 我们还报道了使用盲肠结扎和穿刺模型患全身脓毒症的小鼠的寿命。 这些 ApoE 模拟物穿过血脑屏障，减少多种脑部炎症 阿尔茨海默病模型。 在这项资助中，我们建议通过一种独特的策略优先杀死牙龈卟啉单胞菌。该策略涉及一个。 靶向配体特异性结合牙龈卟啉单胞菌上的独特细胞表面蛋白，当缀合时 与我们的抗菌 ApoE 模拟物一起，提供了一种“靶向”抗菌剂。 靶向基序”作为“gingi-tif”，与 ApoE 模拟物缀合作为“ApoE-gingitif”。 原则上，我们现在证明，被称为 RGN2002 的 ApoE-gingitif 保留了细菌杀灭活性，并且 优先针对杀死牙龈卟啉单胞菌而不是杀死共生细菌（图 9 和表 2）。 合成额外的 ApoE-gingitif 和 gingitif-ApoE 结合物并用它们来测量最小值 我们还将使用这些 ApoE-gingitif 和 gingitif-ApoE。 结合物来测量针对口腔中也发现的非致病性共生细菌的 MIC。 杀死致病性牙龈卟啉单胞菌与非致病性共生菌的 MIC 的比较将提供 优先杀死牙龈卟啉单胞菌的定量，我们称之为“优先比率”（表 2）。 体外筛选已选择合适的 ApoE-gingitif 或 gingitif-ApoE 结合物，然后我们将测试这些 由于口服牙龈卟啉单胞菌导致了整个小鼠牙龈卟啉单胞菌感染模型中的缀合物。 淀粉样蛋白 β 肽 1-42 增加、磷酸化 tau 增加和细胞因子增加，这些都是 在阿尔茨海默病患者的大脑中发现了这些产物，我们将在感染牙龈卟啉单胞菌的大脑中测量这些产物 接受和不接受 ApoE-gingitif 或 gingitif-ApoE 缀合物治疗的小鼠这些结果将使我们能够 确定这种有针对性的“ApoE-gingitif-策略”减少脑部炎症和病理的功效 与 AD 和牙龈卟啉单胞菌感染有关。