Allogeneic and Autologous Immunotherapy for cancer and nonmalignant hematological disorders

癌症和非恶性血液疾病的同种异体和自体免疫疗法

• 批准号: 8939772
• 负责人: RICHARD CHILDS
• 金额: $ 276.74万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8939772
• 关键词: Accounting; Acute Graft Versus Host Disease; Adoptive Immunotherapy; Adoptive Transfer; Adult; Allogenic; Allografting; Animal Model; Animals; Antigen Targeting; Antigens; Antithymoglobulin; Aplastic Anemia; Area; Autologous; Back; Bone Marrow; Bortezomib; CD34 gene; CSF3 gene; Cancer Patient; Cancer Vaccines; Carcinoma; Cells; Cellular Immunity; Cessation of life; Chimerism; Clear Cell; Clinical; Clinical Research; Clinical Trials; Cyclophosphamide; Cytotoxic T-Lymphocytes; DNA; Data; Depsipeptides; Development; Disadvantaged; Disease; Disseminated Malignant Neoplasm; Donor person; Dose; Effector Cell; Electroporation; Engraftment; Enrollment; Equus caballus; Future; Goals; Graft Rejection; HERVs; Hematologic Neoplasms; Hematological Disease; Histone Deacetylase Inhibitor; Homing; Homologous Transplantation; Hour; Human; Hypothyroidism; Hypoxia; Immunity; Immunosuppressive Agents; In Vitro; Infusion procedures; Laboratory Research; Ligands; Macaca mulatta; Malignant - descriptor; Malignant Epithelial Cell; Malignant Neoplasms; Mediating; Messenger RNA; Metastatic Renal Cell Cancer; Methods; Minor; Monoclonal Antibodies; Multiple Myeloma; Multivariate Analysis; Mus; Natural Killer Cells; Neoplasm Metastasis; Neutropenia; Niacinamide; Non-Malignant; Normal tissue morphology; Pancytopenia; Patients; Pharmaceutical Preparations; Phenotype; Population; Proteins; Proviruses; Recovery; Refractory; Regimen; Renal Cell Carcinoma; Renal carcinoma; Reporting; Research; Research Personnel; Risk; Safety; Secondary to; Solid Neoplasm; Source; Specificity; Speed; Stem cell transplant; Stem cells; Surface; Survival Rate; Syndrome; System; T-Lymphocyte; TNFSF10 gene; Technology; Testing; Therapeutic; Thyroiditis; Toxic effect; Transfection; Translational Research; Transplantation; Tumor Antigens; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; VHL Gene Inactivation; Whole-Body Irradiation; Work; base; cancer immunotherapy; cell killing; chronic graft versus host disease; cohort; conditioning; cytotoxicity; demethylation; expression cloning; flasks; fludarabine; follow-up; gene induction; graft vs host disease; improved; in vivo; inhibitor/antagonist; insight; killings; leukemia; lymph nodes; melanoma; multicatalytic endopeptidase complex; neoplastic cell; neutrophil; novel; peripheral blood; phase 1 study; prevent; programs; rat Piga protein; receptor; research study; selective expression; transcription factor; tumor

Approximately 154 years ago, we initiated the first clinical trail investigating allogeneic stem cell transplantation in patients with treatment refractory metastatic renal cell carcinoma (RCC). Definitive evidence for an allogeneic graft-versus-RCC effect has been demonstrated with complete regression of large metastasis observed in some patients. We have subsequently conducted studies investigating the immunological mechanisms accounting for tumor regression in patients demonstrating an anti-RCC effect in attempts to identify both the effector cell populations mediating these regressions as well as their target antigens. We demonstrated that cytotoxic T-cell clones with specificity for minor antigens are capable of killing RCC cells in patients having a GVT effect post transplant. Using c-DNA expression cloning, we have recently identified a novel tumor antigen derived from a human endogenous retrovirus over-expressed in RCC cells called HERV-E CT-RCC. This antigen is not expressed on normal tissues and therefore could potentially serve as a target for a future kidney cancer vaccine. Work to identify other antigens derived from this HERV-E that are expressed on common HLA molecules as well as efforts to develop a monoclonal antibody that recognizes HERV-E derived proteins that are selectively expressed in RCC are ongoing. We have also sought to characterize the mechanisms accounting for selective expression of the CT-RCC HERV-E in RCC. Recently, we found that inactivation of the VHL tumor suppressor gene, over-expression of the hypoxia-inducible transcription factor (HIF) HIF-2, and demethylation of the HERV-E 5LTR results in the selective expression of HERV-E in clear cell carcinoma. These studies have provided valuable insights needed for ongoing translational research aimed at boosting human immunity against antigenic components of this provirus. We are also exploring methods to sensitize solid tumors to NK cell attack by altering the phenotype of tumor cells through targeted gene induction. Recently we showed that bortezomib and depsipeptide sensitize tumors to NK cell cytotoxity by enhancing NK-cell mediated TRAIL killing. This sensitization appears to overcome NK inhibition that is mediated through KIR-KIR ligand interactions. We have also developed a method to expand by > 4 logs NK cells from healthy donors for adoptive infusion in future NK-cell based adoptive immunotherapy trials. We now have developed a method to expand human NK cells by 100 to 1000 fold initially in bags and now in G-REX flasks using GMP conditions. A phase I study entitled Safety and the anti- tumor effects of escalating doses of adoptively infused ex vivo expanded autologous natural killer cells against metastatic cancers or hematological malignancies sensitized to NK TRAIL cytotoxicity with Bortezomib was initiated in 2008 and is currently accruing patients. With the exception of thyroiditis, infusions of up to 2.5x108 expanded NK cells/kg have been well tolerated and provided early evidence for mediating anti-tumor effects in patients with cancer. We have also initiated NK cell studies in rhesus macaques to evaluate methods to optimize the antitumor function and in vivo persistence and tumor homing of adoptively infused autologous NK cells. We have also recently partnered with Dr. Ola Landgren from the NCI to explore KIR blockade with the monoclonal antibody IPH 2101 (Innate Pharma) as a method to enhance autologous NK cell killing of smoldering myeloma. Finally, our lab has recently shown that mRNA electroporation of ex vivo expanded NK cells using the clinical-grade Maxcyte transfection system is highly efficient at transfecting NK cells opening numerous new possibilities to genetically reprogram NK cells to advance the field of NK cell-based cancer immunotherapy. My group has continued to investigate methods to improve allogeneic HCT for heavily pretreated patients with bone marrow failure syndromes (BMFS) including paroxysmal nocturnal hemoglobinuria (PNH) and ATG-refractory severe aplastic anemia (SAA). In a cohort of 56 heavily transfused and HLA alloimmunized patients with BMFS undergoing a novel fludarabine-based HCT, graft rejection did not occur and 87% of the patients survived disease free with a median follow-up of 4.5 years. Despite this excellent survival, approximately 70% of patients developed chronic GVHD. In a multivariate analysis, we found that rapid donor T-cell engraftment (d 95% donor T-cell chimerism by day 30) was associated with the development of chronic GVHD (adjusted HR 3.03, p=0.04). To test whether delaying the speed of donor T-cell engraftment will prevent chronic GVHD, we have initiated a new clinical trial for BMFS patients which uses a modified donor allograft containing high doses of G-CSF mobilized CD34+ selected cells combined with a one log reduced dose of non-G-CSF mobilized T-cells. Preliminary data suggest this transplant approach slows the speed of donor T-cell engraftment compared to a T-cell replete trransplant, significantly reducing the risk of both acute and chronic GVHD Transplantation using unrelated cord blood (UCB) has been shown to be a reasonable alternative transplant approach for patients with a variety of hematological malignancies and non-malignant hematological disorders who lack an HLA matched related or unrelated stem cell donor. Recent data show leukemia-free survival in adult patients with AML and ALL is comparable to that of recipients receiving fully matched unrelated donor transplants. However, for patients with SAA undergoing UCB transplantation, results have been disappointing. Investigators from the EBMT and Eurocord have reported engraftment rates of only 51% and survival rates of only 38% in SAA recipients of single or dual CB transplants. In order to harness the advantage of UCB availability, to overcome the disadvantage of delayed neutrophil recovery of UCB transplantation, and to provide a back up stem cell source should the CB unit fail to engraft, we have recently initiated a clinical trial that evaluates the co-administration of unrelated umbilical cord blood and a relatively low number of highly purified haploidentical peripheral blood CD34+ cells from a related donor as a method to promote rapid neutrophil recovery. Subjects receive a novel non-myeloablative immunosuppressive conditioning regimen of cyclophosphamide, fludarabine, horse ATG and one dose of total body irradiation (200cGy) followed by an infusion of the allografts. The haploidentical stem cell product is T-cell depleted and enriched for CD34+ cells using the Miltenyi CliniMacs system. To reduce TRM secondary to prolonged neutropenia associated with conventional UCB transplantation, haploidentical CD34+ stem cells are co-infused with a single UCB unit (serologically matched at d 4/6 HLA loci). The primary endpoint will be donor engraftment by day 42. This study is now accruing patients with 20/20 intial patients treated having donor engraftment by day 22 and with a median followup of 2.3 years, survival has been excellent at 87%.

大约 154 年前，我们启动了第一个临床试验，研究难治性转移性肾细胞癌 (RCC) 患者的同种异体干细胞移植。在一些患者中观察到的大转移的完全消退已经证明了同种异体移植物抗肾细胞癌效应的明确证据。我们随后进行了研究，调查了患者肿瘤消退的免疫学机制，证明了抗肾细胞癌效应，试图鉴定介导这些消退的效应细胞群及其靶抗原。我们证明，对次要抗原具有特异性的细胞毒性 T 细胞克隆能够杀死移植后具有 GVT 效应的患者的 RCC 细胞。通过 c-DNA 表达克隆，我们最近鉴定了一种新的肿瘤抗原，该抗原源自在 RCC 细胞中过表达的人内源性逆转录病毒，称为 HERV-E CT-RCC。这种抗原在正常组织中不表达，因此有可能作为未来肾癌疫苗的靶标。鉴定源自这种 HERV-E 并在常见 HLA 分子上表达的其他抗原的工作以及开发可识别在 RCC 中选择性表达的 HERV-E 衍生蛋白的单克隆抗体的工作正在进行中。我们还试图描述 RCC 中 CT-RCC HERV-E 选择性表达的机制。最近，我们发现VHL抑癌基因的失活、缺氧诱导转录因子（HIF）HIF-2的过度表达以及HERV-E 5LTR的去甲基化导致了透明细胞癌中HERV-E的选择性表达。这些研究为正在进行的旨在增强人类针对这种原病毒抗原成分的免疫力的转化研究提供了宝贵的见解。 我们还在探索通过靶向基因诱导改变肿瘤细胞表型，使实体瘤对 NK 细胞攻击敏感的方法。最近我们发现硼替佐米和缩酚肽通过增强 NK 细胞介导的 TRAIL 杀伤作用使肿瘤对 NK 细胞的细胞毒性敏感。这种敏化似乎克服了通过 KIR-KIR 配体相互作用介导的 NK 抑制。我们还开发了一种方法，可以将来自健康捐赠者的 NK 细胞扩增超过 4 个对数，用于未来基于 NK 细胞的过继免疫治疗试验中的过继输注。我们现在开发了一种方法，可以使用 GMP 条件将人类 NK 细胞扩增 100 至 1000 倍，最初是在袋子中，现在是在 G-REX 烧瓶中。一项题为“安全性和渐增剂量的体外过继输注扩增自体自然杀伤细胞对抗硼替佐米对 NK TRAIL 细胞毒性敏感的转移性癌症或血液恶性肿瘤的安全性和抗肿瘤作用”的 I 期研究于 2008 年启动，目前正在招募患者。除甲状腺炎外，输注高达 2.5x108 扩增的 NK 细胞/kg 具有良好的耐受性，并为癌症患者介导抗肿瘤作用提供了早期证据。我们还在恒河猴中启动了 NK 细胞研究，以评估优化过继输注自体 NK 细胞的抗肿瘤功能、体内持久性和肿瘤归巢的方法。我们最近还与 NCI 的 Ola Landgren 博士合作，探索使用单克隆抗体 IPH 2101 (Innate Pharma) 阻断 KIR，作为增强自体 NK 细胞杀伤冒烟性骨髓瘤的方法。最后，我们的实验室最近表明，使用临床级 Maxcyte 转染系统对离体扩增的 NK 细胞进行 mRNA 电穿孔，可高效转染 NK 细胞，为对 NK 细胞进行基因重编程以推进基于 NK 细胞的癌症免疫治疗领域开辟了许多新的可能性。 我的团队继续研究改善经过大量治疗的骨髓衰竭综合征 (BMFS) 患者的同种异体 HCT 的方法，这些患者包括阵发性睡眠性血红蛋白尿 (PNH) 和 ATG 难治性严重再生障碍性贫血 (SAA)。在 56 名大量输血且 HLA 同种免疫的 BMFS 患者接受新型基于氟达拉滨的 HCT 的队列中，没有发生移植排斥，并且 87% 的患者在中位随访 4.5 年时无病生存。尽管生存率很高，但大约 70% 的患者出现了慢性 GVHD。在多变量分析中，我们发现供体 T 细胞的快速植入（第 30 天供体 T 细胞嵌合达到 95%）与慢性 GVHD 的发生相关（调整后的 HR 3.03，p=0.04）。为了测试延迟供体 T 细胞植入速度是否会预防慢性 GVHD，我们针对 BMFS 患者启动了一项新的临床试验，该试验使用改良的供体同种异体移植物，其中含有高剂量的 G-CSF 动员的 CD34+ 选择细胞，并结合剂量减少一对数的非 G-CSF 动员的 T 细胞。初步数据表明，与 T 细胞充足移植相比，这种移植方法减慢了供体 T 细胞植入的速度，显着降低了急性和慢性 GVHD 的风险 对于患有各种血液恶性肿瘤和非恶性血液疾病且缺乏 HLA 匹配的相关或无关干细胞供体的患者来说，使用无关脐带血 (UCB) 移植已被证明是一种合理的替代移植方法。最近的数据显示，患有 AML 和 ALL 的成年患者的无白血病生存率与接受完全匹配的无关供体移植的受者的生存率相当。然而，对于接受 UCB 移植的 SAA 患者，结果却令人失望。 EBMT 和 Eurocord 的研究人员报告称，单次或双次 CB 移植的 SAA 受者的植入率仅为 51%，存活率仅为 38%。为了利用 UCB 可用性的优势，克服 UCB 移植中性粒细胞恢复延迟的缺点，并在 CB 单元移植失败时提供备用干细胞来源，我们最近启动了一项临床试验，评估无关脐带血和来自相关脐带血的相对少量的高度纯化的单倍体外周血 CD34+ 细胞的共同给药。 供体作为促进中性粒细胞快速恢复的方法。受试者接受一种新型非清髓性免疫抑制调理方案，包括环磷酰胺、氟达拉滨、马 ATG 和一剂全身照射 (200cGy)，然后输注同种异体移植物。使用 Miltenyi CliniMacs 系统对半相合干细胞产品进行 T 细胞去除和 CD34+ 细胞富集。为了减少继发于传统 UCB 移植相关的长期中性粒细胞减少症的 TRM，将半相合 CD34+ 干细胞与单个 UCB 单位（在 d 4/6 HLA 位点血清学匹配）共同输注。 主要终点将是第 42 天的供体植入。这项研究目前正在招募 20/20 的初始患者，其中 20/20 的患者在第 22 天时接受了供体植入，中位随访时间为 2.3 年，生存率高达 87%。