Off-the-shelf engineered NK cells for the treatment of AML

摘要 冠状病毒（COVID-19）大流行在全球迅速蔓延，感染了300多万人， 对于有严重和危及生命的并发症的患者，没有有效的治疗选择。这样做的影响 在接受造血干细胞移植（HSCT）的患者中，感染可能特别严重， 特别是脐带血移植（CBT）接受者或接受嵌合抗原受体（CAR）的人 T或NK细胞治疗，考虑到其免疫功能低下状态、存在医学合并症和问题 与感染相关的严重程度和死亡率更高。我们的团队已经开发了强大的HLA型临床库， GMP级病毒特异性T淋巴细胞（VST）靶向巨细胞病毒（CMV）、BK病毒（BKV）和 腺病毒。我们已经治疗了100多名HSCT受者，其中49%接受了CB或单倍体相合 移植，这些部分HLA匹配和现成的VST对FDA批准的临床方案， 有效率>80%，无毒性。我们已经成功地应用这个平台建立了 GMP级COVID-19特异性T细胞的生产。我们建议建立一个针对COVID-19的生物库， 使用IRB批准的方案（MDACC）从COVID-19感染中恢复的供体的T细胞 实验室02 -0630）。然后我们将进行I/II期临床试验以评估安全性， 第三方、现成、最接近HLA匹配的COVID-19特异性T细胞的可行性和抗病毒活性 在HSCT中，包括严重COVID-19感染的CBT/单倍体相合移植或细胞治疗接受者（目的 1）。为了加快对我们患者的这项试验的批准，我们最近修改了另一项VST方案， 用COVID-19特异性T细胞治疗（MDACC #2017-0350，IND 17761）。 COVID-19病毒感染最严重的表现之一是急性呼吸窘迫综合征 （急性呼吸窘迫综合征），由于呼吸衰竭，通常需要机械通气和高剂量皮质类固醇治疗。 事实上，越来越多的证据表明，使用皮质类固醇可能会降低COVID-1患者的死亡率。 19相关的ARDS，特别是如果在治疗算法的早期给药。然而，COVID-19特异性T细胞 治疗在这样的患者中不是一种选择，因为皮质类固醇诱导过继转移的T细胞的凋亡， 从而显著地限制了该方法的功效。为了应对这一挑战，我们的团队开发了一个 使用CRISPR-1，在病毒特异性T细胞中抑制糖皮质激素受体（GR）的有效和新颖的策略 Cas9基因编辑核受体亚家族3 C组蛋白1基因（NR 3C 1-编码 GR）。在本建议的目标2中，我们将进行IND使能研究，以生产GMP级 NR 3C 1敲除COVID-19特异性T细胞，为随后的1期试验做准备，将通过 替代来源。鉴于我们在产生临床有效的病毒特异性T细胞方面的记录，我们 我们乐观地认为，我们使用COVID-19特异性T细胞的过继免疫治疗方法将在 HSCT或细胞治疗接受者患有危及生命的COVID-19相关感染。

ABSTRACT The coronavirus (COVID-19) pandemic has spread rapidly and globally to infect over 3 million individuals, with no effective therapeutic options for patients with serious and life-threatening complications. The impact of this infection is likely to be especially serious in patients undergoing hematopoietic stem cell transplant (HSCT), in particular umbilical cord blood transplant (CBT) recipients or those receiving chimeric antigen receptor (CAR) T or NK cell therapies, given their immunocompromised state, presence of medical comorbidities, and concerns for higher infection-related severity and mortality. Our team has developed robust clinical banks of HLA-typed GMP-grade viral-specific T lymphocytes (VSTs) targeting cytomegalovirus (CMV), BK virus (BKV) and adenovirus. We have treated over 100 HSCT recipients, 49% of whom had received CB or haploidentical transplants, with these partially HLA-matched and off-the-shelf VSTs on FDA-approved clinical protocols with a >80% response rate and no toxicity. We have successfully applied this platform to establish the protocols for the manufacture of GMP-grade COVID-19 specific T cells. We propose to generate a biobank of COVID-19 specific T cells from donors who have recovered from COVID-19 infections using IRB-approved protocols (MDACC Lab02-0630) in the MDACC GMP Facility. We will then conduct a phase I/II clinical trial to evaluate the safety, feasibility and antiviral activity of third-party, off-the-shelf, most closely HLA-matched COVID-19 specific T cells in HSCT including CBT/haploidentical transplant or cell therapy recipients with severe COVID-19 infections (Aim 1). To expedite the approval of this trial for our patients, we recently amended another VST protocol to include treatment with COVID-19 specific T cells (MDACC #2017-0350, IND 17761). One of the most severe manifestations of the COVID-19 viral infection is acute respiratory distress syndrome (ARDS), often requiring mechanical ventilation and high dose corticosteroid therapy due to respiratory failure. Indeed, there is increasing evidence that the use of corticosteroids may reduce mortality in patients with COVID- 19 related ARDS, especially if administered early in the treatment algorithm. However, COVID-19 specific T-cell therapy is not an option in such patients as corticosteroids induce apoptosis of adoptively transferred T cells, thus, significantly limiting the efficacy of this approach. To address this challenge, our group has developed an efficient and novel strategy to inactivate the glucocorticoid receptor (GR) in viral-specific T cells, using CRISPR- Cas9 gene editing of the Nuclear Receptor Subfamily 3 Group C Member1 gene (NR3C1- the gene encoding the GR). In Aim 2 of this proposal, we will perform the IND-enabling studies for the production of GMP-grade NR3C1 knockout COVID-19 specific T-cells in preparation for a subsequent phase 1 trial, to be funded through alternative sources. Given our track-record of generating clinically effective viral-specific T-cells, we are optimistic that our approach of adoptive immunotherapy with COVID-19 specific T cells will be successful in HSCT or cell therapy recipients with life-threating COVID-19 related infections.