Characterizing the physicochemical properties of membraneless condensates and its regulation by delta-9-tetrahydrocannabinol in HIV/SIV infection.

表征无膜冷凝物的物理化学性质及其在 HIV/SIV 感染中通过 delta-9-四氢大麻酚的调节。

• 批准号: 10664337
• 负责人: Mahesh Mohan
• 金额: $ 21.8万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10664337
• 关键词: AIDS/HIV problem; Affect; Alzheimer' s Disease; Amyloid fibers; Anti-HIV Agents; Anti-Infective Agents; Anti-Retroviral Agents; Attenuated; Behavior; Biochemical; Biochemistry; Bioenergetics; Biological; Biological Assay; Biological Process; Biology; Blood; Body Fluids; Brain; CCR5 gene; Cell Nucleolus; Cell Nucleus; Cell physiology; Cells; Cellular biology; Centrosome; Cerebrospinal Fluid; Chemicals; Chronic; Classification; Clinical; Complex; Cytoplasm; Cytoplasmic Granules; Data; Data Set; Development; Disease; Dyes; Electron Microscope; Electron Microscopy; Environment; Exposure to; Genetic Transcription; Goals; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-associated neurocognitive disorder; Homeostasis; Human; Individual; Infection; Inflammation; Interruption; Intestinal Content; Knowledge; Lead; Link; Liquid Chromatography; Liquid substance; Macaca; Macaca mulatta; Malignant Neoplasms; Mediating; Membrane; Microscopy; Modeling; Molecular; Molecular Biology; Molecular Target; Morphology; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Normal Range; Nucleic Acids; Outcome Study; Pathogenesis; Pathologic; Peripheral; Pharmaceutical Preparations; Phase; Physiological; Physiological Processes; Plasma; Play; Process; Property; Proteins; Proteomics; RNA; Regulation; Reporting; Resources; Retrieval; Roentgen Rays; Role; SIV; Saliva; Seminal fluid; Shapes; Stress; Structure; Symptoms; Testing; Tetrahydrocannabinol; Thermodynamics; Viral; Viral Pathogenesis; Viral Proteins; Virus; Virus Diseases; Virus Replication; Work; antiretroviral therapy; biological adaptation to stress; cytotoxic; data resource; design; electron energy; experimental study; extracellular vesicles; in vivo; live cell imaging; marijuana use; nanocompound; neuronal transport; new therapeutic target; novel; particle; physical property; protein aggregation; protein complex; protein function; recruit; response; scaffold; stress granule; tat Protein; tool; transmission process

Abstract The `central dogma' of molecular biology is that in cells, the functions of proteins and nucleic acids are primarily determined by their sequence. However, this dogma has been challenged by studies of secreted molecules including the membrane encased extracellular vesicles (EVs) and the membraneless structures known as BMCs, which indicate that cells create discrete chemical environments where they organize complex biomolecules. BMCs form in many compartments of the body, including the nucleus (nucleolus, centrosome, Cajal body, stress granules), the cytoplasm (P-bodies, stress granules), and even the neuronal cytoplasm (neuronal transport granules). While the details of BMC biology is still evolving, it is known that RNA and proteins with intrinsically disordered regions (IDRs) are key factors that influence intracellular composition, size, and morphology of BMCs. The size and composition of BMCs are also thought to be different. BMCs participate in the regulation of various physiological processes, such as modulation of host transcription and stress response. BMCs also play important roles in pathophysiological conditions, including cancer, neurodegenerative diseases, and viral pathogenesis, including modulation of HIV replication. Indeed, it is known that HIV infection results in peripheral and neuro inflammation, and that chronic exposure to THC in the context of HIV in humans or SIV infection in rhesus macaques results in amelioration of disease or symptoms. However, the regulators of HIV disease and THC response is not completely understood. Despite the reported role of BMCs in various conditions, whether or not BMCs regulate the pathogenesis of HIV infection and the intersection of HIV and THC use is unknown. It is also unknown whether BMCs assemble in vivo, where they may play relevant biological roles. There is paucity of information on the presence of BMCs in body fluids (in vivo) and their role in HIV pathogenesis. The reason for this scarcity of information is partly due to lack of tools for isolation of BMCs from body fluids. In preliminary studies, we developed a novel size-guided Particle Purification Liquid Chromatography (PPLC). We used PPLC to isolate and retrieve dye-free BMCs from human semen, rhesus macaque blood plasma and cerebrospinal fluid (CSF). Using standard electron microscopy and Energy Dispersive X-ray equipped transmission electron microscope (TEM-EDX) analyses, we obtained structural and elemental information of the BMCs. Proteomics analysis and RNA bioanalyzer study were used to show that the BMCs contain protein and RNA, while cell biology assay indicate that BMCs are cytotoxic to cells. These data provide compelling evidence that bioactive BMCs are present in vivo. Thus, in this proposal, we will use PPLC to isolate BMCs from rhesus macaque blood, CSF, saliva, and intestinal contents. We will then use various structural, biochemical, and cell biology assays to provide structural, molecular, and functional classification of BMCs from SIV infected and THC treated macaques, which will provide a framework to dissect BMC functions in HIV/AIDS and THC use.

抽象的 分子生物学的“中央教条”是在细胞中，蛋白质和核酸的功能主要是 由它们的序列确定。但是，该教条受到分泌分子的研究的挑战 包括膜包含的细胞外囊泡（EV）和称为BMC的膜结构， 这表明细胞创建离散的化学环境，它们在其中组织复杂的生物分子。 BMC在身体的许多隔室中形成 颗粒），细胞质（p-bodies，应激颗粒），甚至神经元细胞质（神经元转运 颗粒）。虽然BMC生物学的细节仍在发展，但已知RNA和蛋白质本质上 无序区域（IDR）是影响BMC的细胞内组成，大小和形态的关键因素。 BMC的大小和组成也被认为是不同的。 BMC参与各种监管 生理过程，例如调节宿主转录和应力反应。 BMC也发挥重要作用 在病理生理状况中的作用，包括癌症，神经退行性疾病和病毒发病机理， 包括调节HIV复制。确实，众所周知，HIV感染导致周围和神经 炎症，以及在人类艾滋病毒或恒河神感染的艾滋病毒中长期暴露于THC 猕猴会改善疾病或症状。但是，HIV疾病和THC的调节因子 响应尚未完全理解。尽管据报道BMC在各种条件下的作用，无论是否 BMC调节HIV感染的发病机理，HIV和THC使用的相交尚不清楚。也是 尚不清楚BMC是否会在体内组装，它们可能扮演相关的生物学角色。很少 有关体液中BMC的存在（体内）及其在HIV发病机理中的作用的信息。原因 这种信息的稀缺部分部分是由于缺乏从体液中隔离BMC的工具。在初步 研究，我们开发了一种新型的尺寸引导的颗粒纯化液相色谱（PPLC）。我们使用了PPLC 从人精液，猕猴血浆和脑脊液中分离和检索无染料的BMC 流体（CSF）。使用标准电子显微镜和配备X射线的能量分散透射电子 显微镜（TEM-EDX）分析，我们获得了BMC的结构和元素信息。蛋白质组学 分析和RNA生物分析仪研究表明BMC含有蛋白质和RNA，而细胞 生物学测定法表明BMC对细胞具有细胞毒性。这些数据提供了令人信服的证据表明生物活性 BMC存在于体内。因此，在此提案中，我们将使用PPLC将BMC与猕猴的血液分离出来， CSF，唾液和肠内容物。然后，我们将使用各种结构，生化和细胞生物学测定法 提供来自感染和THC处理的SIV的BMC的结构，分子和功能分类 猕猴，它将提供一个框架，以剖析艾滋病毒/艾滋病和THC使用中的BMC功能。

项目成果

A multicentre study reveals dysbiosis in the microbial co-infection and antimicrobial resistance gene profile in the nasopharynx of COVID-19 patients.

• DOI: 10.1038/s41598-023-30504-3
• 发表时间: 2023-03-13
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Mahmud, A. Sayeed. M.;Seers, Christine A.;Shaikh, Aftab Ali;Taznin, Tarannum;Uzzaman, Mohammad Samir;Osman, Eshrar;Habib, Md. Ahashan;Akter, Shahina;Banu, Tanjina Akhtar;Sarkar, Md. Murshed Hasan;Goswami, Barna;Jahan, Iffat;Okeoma, Chioma M.;Khan, Md. Salim;Reynolds, Eric C.
• 通讯作者: Reynolds, Eric C.

The Past, the Present, and the Future of the Size Exclusion Chromatography in Extracellular Vesicles Separation.

• DOI: 10.3390/v13112272
• 发表时间: 2021-11-13
• 期刊: Viruses
• 作者: Kaddour H;Tranquille M;Okeoma CM
• 通讯作者: Okeoma CM