Allogeneic Immunotherapy for cancer and nonmalignant hematological disorders

癌症和非恶性血液病的同种异体免疫疗法

• 批准号: 8344766
• 负责人: RICHARD CHILDS
• 金额: $ 269.38万
• 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/8344766
• 关键词: Accounting; Activated Natural Killer Cell; Adoptive Immunotherapy; Adoptive Transfer; Adult; Aftercare; Allogenic; Allografting; Animal Model; Animals; Antigen Targeting; Antigens; Antithymoglobulin; Aplastic Anemia; Area; Autologous; Back; Blood specimen; Bortezomib; CD34 gene; CSF3 gene; Cancer Patient; Cancer Vaccines; Carcinoma; Cell Line; Cells; Cellular Immunity; Cessation of life; Chimerism; Clear Cell; Clinical; Clinical Research; Clinical Trials; Correlative Study; Cyclophosphamide; Cytoplasmic Granules; Cytotoxic T-Lymphocytes; DNA; Data; Depsipeptides; Development; Diagnosis; Disadvantaged; Disease; Disseminated Malignant Neoplasm; Donor person; Dose; Effector Cell; Engraftment; Enrollment; Equus caballus; Future; Goals; Graft Rejection; Graft vs Tumor Effect; HERVs; Hematologic Neoplasms; Hematological Disease; Hematopoietic Stem Cell Transplantation; Histone Deacetylase Inhibitor; Homologous Transplantation; Hour; Human; Hypothyroidism; Hypoxia; Immunity; Immunosuppressive Agents; In Vitro; Infusion procedures; Laboratories; Laboratory Research; Ligands; Macaca mulatta; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Mediating; Metastatic Renal Cell Cancer; Methods; Minor; Modeling; Monoclonal Antibodies; Multiple Myeloma; Multivariate Analysis; Mus; Natural Killer Cells; Neoplasm Metastasis; Neutropenia; 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; Safety; Secondary to; Small Interfering RNA; Solid Neoplasm; Source; Specificity; Speed; Stem cell transplant; Stem cells; Surface; Survival Rate; Syndrome; System; T-Lymphocyte; TNFSF10 gene; Techniques; Technology; Testing; Therapeutic; Thyroiditis; Toxic effect; Translating; Translational Research; Transplantation; Tumor Antigens; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; VHL Gene Inactivation; Whole-Body Irradiation; Work; base; cancer cell; cancer immunotherapy; cell killing; chronic graft versus host disease; cohort; conditioning; cytokine; cytotoxic; cytotoxicity; demethylation; expression cloning; fludarabine; follow-up; gene induction; graft vs host disease; improved; in vivo; inhibitor/antagonist; insight; killings; knock-down; leukemia; melanoma; multicatalytic endopeptidase complex; neoplastic cell; neutrophil; novel; overexpression; peripheral blood; phase 1 study; prevent; programs; rat Piga protein; receptor; research study; selective expression; transcription factor; tumor

Approximately 10 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. In order to explore methods to enhance GVT effects against RCC, we have developed a murine model of allogeneic HCT. This model demonstrates that adoptively transferred NK cells from MHC matched donors reduce GVHD and mediate a GVT effect against murine RCC prolonging survival. These findings are being translated into a clinical trial that will explore the potential of ex-vivo expanded donor NK cell infusions to reduce GVHD and enhance GVT in patients with RCC and other cancers undergoing allogeneic HCT. 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 in bags 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 1x108 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 of adoptively infused autologous NK cells.We have also recently shown that siRNA inactivation of NKG2A enhances NK cell tumor cytotoxicity in vitro and in vivo in tumor-bearing mice, a finding that may have therapeutic implications for humans with cancer. Finally, we have 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. Patients with a confirmed diagnosis of smoldering myeloma will receive IPH2101 (1mg/kg) every other month for 6 cycles. Patients will be restaged monthly pre- and post-treatment. In vitro correlative studies from blood samples collected from patients to measure the effects of KIR blockade on NK cell cytotoxic granule release, NK cell cytotoxicity and NK cell intracellular cytokine secretion against myeloma cell lines will be performed in my laboratory. Using this technique, the percentage change in NK cell cytotoxic granule release and NK cell intracellular cytokine secretion following treatment with IPH2101 compared to baseline will be determined and correlated with clinical evidence for disease activity. This study is actively enrolling patients. 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. 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.

大约10年前，我们开始了第一个临床跟踪，研究了治疗难治性转移性肾细胞癌（RCC）患者的同种异体干细胞移植。通过某些患者观察到的大型转移的完全消退，已经证明了同种异体移植物-RCC效应的确定证据。随后，我们进行了研究，研究了对患者肿瘤回归的免疫学机制，该研究证明了抗RCC效应，以识别介导这些回归及其靶抗原的效应细胞群体。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。使用C-DNA表达克隆，我们最近确定了一种新型的肿瘤抗原，该肿瘤抗原源自人内源性逆转录病毒，在RCC细胞中过表达，称为HERV-E CT-RCC。该抗原未在正常组织上表达，因此有可能成为未来肾癌疫苗的靶标。努力鉴定从该HERV-E中得出的其他抗原，这些抗原在共同的HLA分子上表达，并努力开发一种单克隆抗体，该抗体识别出在RCC中选择性表达的Herv-E衍生蛋白的努力正在进行中。我们还试图表征RCC中CT-RCC HERV-E选择性表达的机制。最近，我们发现VHL肿瘤抑制基因的失活，缺氧诱导转录因子（HIF）HIF-2的过表达以及HERV-E 5LTR的去甲基化导致透明细胞癌中HERV-E的选择性表达。这些研究为正在进行的转化研究提供了宝贵的见解，旨在促进对该病毒的抗原成分的人类免疫力。为了探索增强对RCC的GVT效应的方法，我们开发了同种异体HCT的鼠模型。该模型表明，从MHC匹配的供体中采用的NK细胞降低了GVHD，并介导GVT效应，以针对鼠RCC延长生存率。 这些发现正在转化为一项临床试验，该试验将探讨Ex-Vivo扩展的供体NK细胞输注的潜力，以减少RCC和其他正在接受同种异体HCT的癌症患者的GVHD和增强GVT。 我们还正在探索通过通过靶向基因诱导改变肿瘤细胞的表型来使实体瘤对NK细胞攻击的方法。最近，我们表明硼替佐米和深度肽通过增强NK细胞介导的越野杀伤来使肿瘤对NK细胞细胞毒素敏感。这种敏化似乎克服了通过KIR-KIR配体相互作用介导的NK抑制作用。我们还开发了一种方法，可以在未来的基于NK细胞的收养免疫疗法试验中从健康供体中扩展> 4 logs NK细胞，从而从健康的供体中进行收养。现在，我们已经开发了一种使用GMP条件在袋中扩展人类NK细胞100到1000倍的方法。一项I阶段的研究，名为“安全性和抗肿瘤的抗肿瘤作用”的抗肿瘤效应升级的剂量扩大了自体杀伤细胞针对转移性癌症或对NK Trail trail trail trail用bortezomib敏感的抗血压恶性肿瘤，并于2008年开始启动，目前正在培养患者。除甲状腺炎外，多达1x108的NK细胞/kg的输注已得到很好的耐受性，并提供了早期证据，以介导癌症患者的抗肿瘤作用。 We have also initiated NK cell studies in rhesus macaques to evaluate methods to optimize the antitumor function and in vivo persistence of adoptively infused autologous NK cells.We have also recently shown that siRNA inactivation of NKG2A enhances NK cell tumor cytotoxicity in vitro and in vivo in tumor-bearing mice, a finding that may have therapeutic implications for humans with cancer.最后，我们最近与NCI的Ola Landgren博士合作，使用单克隆抗体IPH 2101（先天性药物）来探索KIR封锁，以增强自体NK NK细胞杀死湿透的骨髓瘤的方法。确定湿透骨髓瘤的确认诊断患者每隔一个月将接受6个周期的IPH2101（1mg/kg）。患者将每月在治疗前和治疗后重新及时。从患者中收集的血液样本中的体外相关研究，以测量KIR阻断对NK细胞毒性颗粒释放，NK细胞细胞毒性和NK细胞细胞内细胞因子对骨髓瘤细胞系的分泌的影响。使用该技术，与基线相比，将在IPH2101治疗后NK细胞细胞毒性颗粒释放和NK细胞内细胞因子分泌的百分比变化将被确定并与疾病活性的临床证据相关。这项研究正在积极招募患者。 我的小组继续研究改善经过预处理的骨髓衰竭综合征（BMF）患者的同种异体HCT方法，包括阵发性夜间夜间血红蛋白尿（PNH）和ATG反复严重的严重性性动态性障碍（SAA）。在一组56个经历了新型基于氟达拉滨的HCT的BMF患者的56例重量输血和HLA同染色的患者中，没有发生移植物排斥反应，并且有87％的患者在中位随访的中位随访中幸存下来。尽管生存率出色，但大约70％的患者发展了慢性GVHD。在多元分析中，我们发现快速供体T细胞植入（D 95％的供体T细胞嵌合在第30天）与慢性GVHD的发展有关（调整后的HR 3.03，P = 0.04）。为了测试延迟供体T细胞植入速度的速度是否可以预防慢性GVHD，我们已经为BMFS患者启动了一项新的临床试验，BMFS患者使用含有改良的供体同种异体移植物，其中含有高剂量的G-CSF动员CD34+选定细胞，与非G-CSF动员T-Cells的剂量降低了一log剂量减少剂量。 对于患有多种缺乏HLA匹配的相关或不相关的干细胞供体的患者，使用无关的脐带血（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，AML成年患者无白血病生存期，所有这些都与接受完全匹配的无关供体移植的接受者相当。但是，对于接受UCB移植的SAA患者，结果令人失望。来自EBMT和EuroCord的研究人员报告说，单次或双CB移植的SAA接受者的植入率仅为51％，存活率仅为38％。为了利用UCB可用性的优势，克服UCB移植延迟嗜中性粒细胞恢复的缺点，如果CB单元未植入CB单位，我们最近启动了一项临床试验，该试验启动了一项临床试验，该试验评估了与无关的无关脉冲血液和高度纯粹的纯索尔的同时，CRESID SLOCTAIL ADMERTAIL ADMERAL ADMERTIAL ALMIDICAL ADLICER CLELICAL SPLICER CLELICER pLOLICER cOD级别的纯度数量相关的小数不是，CD供体作为促进快速中性粒细胞恢复的方法。受试者获得了一种新型的非毛囊免疫抑制条件，对环磷酰胺，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系统将单倍同化干细胞产物耗尽并富含CD34+细胞。为了减少继发于常规UCB移植相关的长时间中性粒细胞减少症的TRM，单倍型CD34+干细胞与单个UCB单元共同介绍（在D 4/6 HLA基因座的血清学匹配）。 主要终点将是第42天的捐助者植入。这项研究现在正在累积患者。