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.

翻译后摘要：载脂蛋白E在AD的细菌理论的治疗作用 牙龈卟啉单胞菌是牙周病的关键细菌，牙周病是第6大常见的感染性疾病 国际吧一半的30岁以上的美国人和70%的65岁以上的人患有某种程度的P。 与牙龈疾病相关的牙龈感染。在一项针对6800名牙龈疾病患者的研究中， 在长达26年的随访中，那些患牙龈炎，然后患牙周病的人与 显著增加痴呆症的风险，包括阿尔茨海默氏痴呆症。牙齿脱落，一个常见的结果， 牙周病，也与痴呆症的风险增加有关。牙龈卟啉单胞菌产品 包括脂多糖、称为牙龈卟啉菌蛋白酶的半胱氨酸蛋白酶和来自牙龈卟啉单胞菌的16 S rRNA， 都是在老年痴呆症患者的大脑中发现的。由于牙龈卟啉单胞菌是在口腔中发现的， 已知会侵入包括大脑在内的非口腔组织，那么减少牙龈卟啉单胞菌数量的策略， 活动，包括减少大脑炎症，开始显示出积极的影响， 在动物模型和临床上的应用。 我们最近发现，载脂蛋白-E的小模拟物（ApoE-模拟物）抑制生长 杀死牙龈卟啉单胞菌其他研究小组也报道了类似的ApoE模拟物的抗菌活性 包括我们的ApoE模拟物这些抗菌活性似乎与抗炎作用相关。 我们和其他小组报道的ApoE模拟物的活性。我们报道了这些ApoE模拟物 使用盲肠结扎和穿刺模型，在经受全身脓毒症的小鼠中观察寿命。我们还报道 这些ApoE模拟物穿过血脑屏障，在多个脑组织中减少脑炎症， 阿尔茨海默病的模型。 在这项资助中，我们提出了一种独特的策略，优先杀死牙龈卟啉单胞菌。这一战略涉及一个 靶向配体，其特异性结合牙龈卟啉单胞菌上的独特细胞表面蛋白， 与我们的抗菌ApoE模拟物一起，提供了一种“靶向”抗菌剂。我们称之为“牙龈卟啉单胞菌 靶向基序”作为“gingi-tif”，与ApoE-模拟物缀合作为“ApoE-gingitif”。作为证据- 原则上，我们现在表明，被称为RGN 2002的ApoE-gingitif保留了细菌杀伤活性， 与杀死牙龈细菌相比，优先靶向牙龈卟啉单胞菌杀死（图9和表2）。我们将 合成另外的ApoE-gingitif和gingitif-ApoE缀合物，并使用它们来测量最小 抑制浓度（MIC）。我们还将使用这些ApoE-gingitif和gingitif-ApoE 结合物来测量针对也在口腔中发现的非致病性口腔细菌的MIC。之比 杀死致病性牙龈卟啉单胞菌与非致病性牙龈细菌的MIC之比将提供 对优先杀死牙龈卟啉单胞菌的定量，我们称之为“优先比”。（表2）。一旦我们在 体外筛选选择了合适的ApoE-牙龈炎或牙龈炎-ApoE结合物，然后我们将测试这些 在牙龈卟啉单胞菌感染的整个小鼠模型中，由于牙龈卟啉单胞菌的口服递送导致 增加的淀粉样β肽1-42，增加的磷酸化tau蛋白和增加的细胞因子，这些都是 我们将在被牙龈卟啉单胞菌感染的大脑中测量这些产物 用和不用ApoE-gingitif或gingitif-ApoE缀合物处理的小鼠。这些结果将使我们能够 确定这种靶向“ApoE-gingitif-策略”减少脑炎症和病理学的功效 与AD和牙龈卟啉单胞菌感染有关。