Multivirus-specific T Cells from Naive CB-derived T Cells

项目总结 巨细胞病毒(CMV)、EB病毒(EBV)、腺病毒(Ad)和BK病毒(BKV)引起的病毒感染 仍然是接受异基因造血干细胞治疗失败的重要原因 移植(HSCT)。艾滋病毒携带者因潜在的恶性肿瘤而需要移植的人是 病毒反弹的风险也很高。未感染病毒的捐献者脐带血(CB)或造血干细胞的接受者在 尤其危险，因为他们的移植物不包含病毒特异性记忆T细胞。抗病毒药物仅对以下患者有效 有些病毒，而且大多数都有显著的毒性。病毒特异性细胞毒性T淋巴细胞的过继转移 来自干细胞捐赠者的(VSTs)已被证明是安全和高效的，但通常仅适用于 接受来自有病毒经验的捐赠者的移植，从而排除了风险最高的患者。这种缺乏一种 激活和扩增来自幼稚捐赠者来源(如CB)的病毒特异性T细胞的有效策略已经被 将这一办法推广到高风险接受者的一个主要障碍是，其持续的长期发病率和 病毒性疾病的高死亡率大大降低了移植手术的成本：收益比。在 在上一个融资周期，我们开发了一种新的方法，有效地扩展了特定于(CMV)、(EBV)的VST 和(Ad)来自幼稚的脐带血来源的T细胞。针对三种病毒的CB-VST具有广泛的表位特异性， 安全有效地预防和/或治疗儿童单一脐血后病毒感染12例 移植(CBT)。为了扩大这一方法的适用性，我们现在建议进行以下研究：(I)扩大这一范围 应对其他病毒(如BKV和HIV)的方法，(Ii)采用更快速的制造方案和(Iii) 评价成人双侧非亲缘CBT(DUCBT)后的VST治疗。因此，我们现在假设， 在CBT之后，快速制造的针对四种病毒(CMV，EBV，Ad， BKV)将是安全的(目标1)，并在早期(&lt；60天)提供广泛的保护，防止在- DUCBT(目标2)。我们进一步假设，这种方法对于启动HIV特异性T细胞是有效的。 来自CB的细胞，潜在地作为CBT后的治疗策略(目标3)。我们产生的结果将表明 这一新策略可以持续产生针对最常见致病病毒的有效VST 包括艾滋病毒在内的CBT，以及这种方法是否有可能降低术后的发病率和死亡率 移植手术。在上一个资金周期中，我们展示了采用转让的安全性和可行性 为接受CBT的儿科患者提供VST为与项目1和项目3的合作奠定了基础 (在CB扩增/归巢和CB-NK细胞的临床试验中，纵向T细胞重建分别为 岩藻糖基化)以及项目4，其嵌合抗原受体靶向方法可用于 对VST进行基因改造，使其具有传统病毒表位的特异性。

PROJECT SUMMARY Viral infections due to cytomegalovirus (CMV), Epstein-Barr virus (EBV), adenovirus (Ad) and BK virus (BKV) remain a significant cause of treatment failure in patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT). Individuals living with HIV who require a transplant for an underlying malignancy are also at high risk for viral rebound. Recipients of cord blood (CB) or HSC from virus-naïve donors are at particular risk since their grafts contain no virus-specific memory T-cells. Antiviral drugs are effective only for some viruses, and most have significant toxicities. Adoptive transfer of virus-specific cytotoxic T lymphocytes (VSTs) from the stem cell donor has proved safe and highly effective, but has generally only been for recipients of grafts from virus-experienced donors, thereby excluding patients at highest risk. This lack of an effective strategy to activate and expand virus-specific T-cells from naïve donor sources such as CB, has been a major obstacle to extending the approach to high-risk recipients, whose continued prolonged morbidity and high mortality from viral diseases substantially reduces the cost:benefit ratio of the transplant procedure. In the last funding cycle, we developed a novel approach that effectively expanded VSTs specific for (CMV), (EBV) and (Ad) from naïve CB-derived T-cells. CB-VSTs targeting three viruses had broad epitope specificity, were safe and effectively prevented and/or treated viral infections in 12 pediatric patients after single cord blood transplantation (CBT). To broaden the applicability of this approach, we now propose studies to: (i) extend this approach to additional viruses (e.g. BKV and HIV), (ii) employ a more rapid manufacturing protocol and (iii) evaluate VST therapy in adult patients after double unrelated CBT (DUCBT). Hence, we now hypothesize that, following CBT, the infusion of rapidly manufactured CB-derived VSTs targeting FOUR viruses (CMV, EBV, Ad, BKV) will be safe (Aim 1) and provide broad protection early (< 60 days) against viral infections arising post- DUCBT (Aim 2). We further hypothesize that this approach will be effective for the priming of HIV-specific T- cells from CB, potentially as a curative strategy post-CBT (Aim 3). The results we generate will show whether this novel strategy can consistently produce effective VSTs directed to the commonest pathogenic viruses after CBT, including HIV, and whether this approach has the potential to reduce morbidity and mortality after transplantion. Our demonstration, during the last funding cycle, of the safety and feasibility of adoptive transfer of VSTs to pediatric patients undergoing CBT set the stage for collaborations with Projects 1 and 3 (longitudinal T-cell reconstitution in clinical trials of CB expansion/homing and of CB-NK cells, repectively and fucosylation) as well as Project 4, whose chimeric antigen receptor targeting approach could be used to genetically modify VSTs to render them specific for conventional virus epitopes.