Developing an NHP model for understanding the biological causes of long COVID-19 pathogenesis

摘要 截至2021年4月30日，新冠肺炎大流行已导致超过1.48亿例和310万例 世界各地的死亡人数。通常情况下，人们会在2到6周后从新冠肺炎康复；然而，在相当大一部分时间里 部分患者的症状可能会在最初康复后的几周或几个月内持续或复发。这些“长新冠肺炎” SARS-CoV-2感染(PASC)的症状或急性后遗症包括疲劳或肌肉无力、睡眠 失调、味觉或嗅觉丧失、困惑、焦虑和抑郁。而临床特点和 急性新冠肺炎病的发病机制正在深入研究，该病的长期后果 在很大程度上仍然不为人知。此外，迄今为止发表的少量PASC研究报告也是有限的 患者出院后的随访时间相对较短；缺乏感染前数据；以及 有限的组织采样。在这里，利用最近建立的非人灵长类(NHP)模型 SARS-CoV-2感染，我们的目标是建立生物安全指南来验证和标准化PASC NHP 对恒河猴的研究--感染后长达18周--动物反复检测结果为阴性 SARS-CoV-2将从ABSL-3转移到ABSL-2+设施(目标1)。这种将动物从 ABSL-3设施将是关键，以便能够以可接受的成本和劳动力在NHP中研究PASC，而 保护人员和NHP殖民地的安全。将动物转移到BSL-2+将使我们能够 描述在人类PASC中观察到的神经和行为表现。此外，我们正在 在对SARS冠状病毒的长期研究中提出广泛和最先进的免疫学和病毒学分析- 2感染恒河猴(RMS)以确定PASC的发病机制和机制 (目标2)。重要的是，我们的研究将有助于建立SARS-CoV-2感染的NHP模型和 可能被证明是理解SARS-CoV-2致病机制的长期影响的关键。该模型提供了 对疾病发现、发病机制、免疫反应和病毒持久性进行纵向评估。 此外，收集人类几乎不可能获得的多种组织，如肠道、大脑、心脏和 肺，将使我们能够识别肺和肺外的长期和/或永久性损害。最后， 纵向和尸检收集的标本将被冷藏库，这将是连接我们发现的关键 数据汇编在SARS-CoV-2恢复期人类队列中。这些研究将使我们能够解剖生物学 PASC的潜在原因，从而为预防和治疗长期COVID的影响提供了关键的见解。 19种疾病。拟议的研究属于家长奖励的范围，极有可能促进 额外的研究经费，使家长奖的总体目标取得进展。然而，还有 与已在父奖项中资助的工作没有重叠。

Abstract As of April 30, 2021, the COVID-19 pandemic has resulted in more that 148 million cases and with 3.1 million deaths worldwide. Typically, people recover from COVID-19 after 2 to 6 weeks; however, in a significant fraction of patients symptoms may linger or recur for weeks or months following initial recovery. These “long COVID-19” symptoms or Post-Acute Sequelae of SARS-CoV-2 infection (PASC) include fatigue or muscle weakness, sleep disorders, loss of taste or smell, confusion, anxiety, and depression. While the clinical characteristics and pathogenesis of acute COVID-19 disease are being intensively studied, the long-term consequences of disease remain largely unknown. Furthermore, the small number of PASC research studies published to date are limited by a relatively short follow-up after patients are discharged from the hospital; a lack of pre-infection data; and limited sampling of tissues. Here, taking advantage of a recently established nonhuman primate (NHP) model of SARS-CoV-2 infection, we aim to establish biosafety guidelines to validate and standardize PASC NHP studies in rhesus macaques- up to 18 weeks post-infection – in which animals repeatedly testing negative for SARS-CoV-2 will be transferred from ABSL-3 to ABSL-2+ facilities (Aim 1). This ability to transfer animals from ABSL-3 facilities will be critical to allow the study of PASC in NHP with acceptable costs and labor, while preserving the safety of personnel and the NHP colonies. Transfer of animals to BSL-2+ will allow us to characterize neurological and behavioral manifestations observed in human PASC. Additionally, we are proposing extensive and state-of-the-art immunologic and virologic analyses in a long-term study of SARS-CoV- 2-infected rhesus macaques (RMs) to define the pathogenesis and identify mechanisms underlying PASC (Aim 2). Importantly, our study will contribute to the establishment of NHP models of SARS-CoV-2 infection and could prove essential for understanding long-term effects of SARS-CoV-2 pathogenesis. This model provides longitudinal assessment of disease findings, pathogenesis, immune responses, and viral persistence. Furthermore, collection of multiple tissues virtually impossible to obtain in humans, such as gut, brain, heart and lung, will allow us to identify pulmonary and extra-pulmonary long-term and/or permanent damage. Finally, specimens collected longitudinally and at necropsy will be cryo-banked and will be key in bridging our discoveries with data compiled in SARS-CoV-2 recovery human cohorts. These studies will allow us to dissect biological causes underlying PASC, thus providing key insights for preventing and treating the effects of long-term COVID- 19 disease. The proposed studies are within the scope of the parent award and are highly likely to foster additional research funding leading to the progress of the overall goals of the parent award. However, there is no overlap with work already funded in the parent award.