Targeting the Epigenome for Lung Cancer Therapy

针对肺癌治疗的表观基因组

• 批准号: 8938407
• 负责人: DAVID SCHRUMP
• 金额: $ 47.65万
• 依托单位: DIVISION OF CLINICAL SCIENCES - NCI
• 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/8938407
• 关键词: Aberrant DNA Methylation; Acute Erythroblastic Leukemia; Adjuvant; Adjuvant Therapy; Adoptive Transfer; Aftercare; Antigens; Apoptosis; Attenuated; Autologous; Biopsy; Buffers; CTAG1 gene; Cancer Patient; Cell Cycle Progression; Cell Line; Cells; Chest; Chromatin Structure; Chromosome Mapping; Clinical; Clinical Protocols; Cyclophosphamide; DNA; DNA Methylation; Data; Decitabine; Depsipeptides; Derivation procedure; Disease; Endogenous Retroviruses; Epigenetic Process; Evaluation; Event; Excision; Exhibits; Freezing; Gene Expression; Genes; Genomic Instability; Germ Cells; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; H1299; Histology; Histone Deacetylase Inhibitor; Histones; Human; Immune response; Immunization; Immunocompetent; Immunologics; Immunosuppressive Agents; Individual; Infusion procedures; Intervention; K-562; K562 Cells; Laboratories; Lasers; Life; MAGEA3 gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of testis; Manuscripts; Mediating; Metastatic to; Modeling; Modification; Molecular; Mus; Neoplasm Metastasis; Oncogenes; Oral; Patients; Phase II Clinical Trials; Polycomb; Positioning Attribute; Primary Neoplasm; Proteins; Publications; Publishing; Randomized; Recurrence; Regimen; Relative (related person); Repression; Resected; Series; Serological; Site; Structure of respiratory epithelium; T-Lymphocyte; Thoracic Oncology; Thoracic Surgical Procedures; Toxic effect; Tumor Suppressor Genes; Tumor Tissue; Up-Regulation; Vaccination; Vaccine Adjuvant; Vaccines; Whole Cell Vaccine; X Chromosome; bronchial epithelium; cancer cell; cancer immunotherapy; cancer stem cell; cancer therapy; celecoxib; chemotherapy; chromatin remodeling; conventional therapy; demethylation; epigenome; genome-wide; high risk; immunogenic; immunogenicity; imprint; in vivo; meetings; neoplastic cell; novel; pre-clinical; preclinical study; research study; response; senescence; therapeutic vaccine; therapy development; tumor; vaccine development; vaccine-induced immunity

During the past decade, more than 100 patients with thoracic malignancies have been treated on a series of clinical protocols examining toxicities and clinical responses following infusions of DNA demethylating agents ( Decitabine; DAC), and HDAC inhibitors, such as romidepsin (DP). Collectively these trials demonstrated no objective clinical regressions, although prolonged stabilization of disease (4-12months) was observed in approximately 10% of patients. Nearly one quarter of all patients receiving DAC infusions exhibited increased expression of p16, MAGE-3, or NY-ESO-1 in tumor tissues. Serologic responses to NY-ESO-1 were observed in several patients receiving DAC for more than six months. Approximately 50% of patients receiving DP infusions exhibited increased intratumoral levels of H3Ac and p21. In addition, several patients exhibited enhanced expression of NY-ESO-1 and MAGE-A3 in tumor biopsies following DP infusions. Micro-array analysis of laser captured tumor cells from pre and post treatment biopsies from patients receiving DAC, DP or sequential DAC/DP infusions revealed a shift from a lung cancer gene signature to one observed in normal respiratory epithelia. These findings, together with observations of up-regulation of CT-X antigens in tumor tissues provided proof of concept for the use of epigenetic regimens in combination with immunologic interventions to target CT-X antigens in thoracic malignancies. In general, thoracic malignancies exhibit low level, focal and heterogeneous CT-X gene expression, as well as variable responses to chromatin remodeling agents. Because CT-X antigens appear to be preferentially expressed in pluripotent tumor cells, it is conceivable that autologous epigenetically modified tumor cells may be unique, personalized vaccines targeting cancer stem cells, which promote systemic metastases. Recent clinical efforts have focused on the evaluation of epigenetically-modified tumor cells as adjuvant vaccines in patients with primary thoracic malignancies, as well as patients with extra-thoracic malignancies metastatic to the chest, who have been rendered NED by standard therapy. The vaccines are administered with Iscomatrix, a proprietary adjuvant that has been shown to elicit potent immunologic responses to purified CT-X antigens; additionally, oral celecoxib will be used to inhibit activity of Tregs that have been shown to attenuate immune responses to therapeutic vaccines in cancer patients. To date, 30 patients have been accrued to this trial. Despite preclinical studies demonstrating feasibility of this approach, reliable derivation of cell lines from primary tumors has been a significant challenge, in part due the histologies of the resected malignancies. Although cells lines have been established from several individuals, these patients were ineligible for vaccinations due to disease recurrence. In an attempt to circumvent the problems of establishing primary cell lines, we have identified several established lines including K562 erythroleukemia, and H1299 lung cancer cells that exhibit high level CT-X gene expression without pharmacologic manipulation for use as adjuvant vaccines. In a recent Phase II trial in the Thoracic Surgery Section, TGIB, 20 patients received six, monthly immunizations with 2.5e7-1e8 live irradiated K562 cells constitutively expressing GM-CSF (K562-GM) in conjunction with metronomic oral cyclophosphamide and celecoxib (to inhibit immunosuppressive Tregs) as adjuvant therapy following complete resection of their malignancies. Metronomic chemotherapy was well tolerated, and no vaccine-related toxicities were observed. Whereas several patients exhibited increased serologic reactivity to purified CT-X antigens following their vaccinations, none met criteria for immunologic response. Several patients with uncharacteristically long disease free intervals while on therapy, developed rapid recurrence shortly after being taken off study. These findings raise the possibility that there were responses to the vaccine that were not detected by our analysis, and/or a potential clinical benefit of the metronomic chemotherapy. A manuscript pertaining to this trial is presently being prepared for publication. In more recent efforts we have developed a vaccine using H1299 lung cancer cells. Due to amplification of the X chromosome, H1299 cells exhibit broader and higher levels of CT-X gene expression than K562-GM; as such, H1299 cells may be more effective vaccines for inducing immunity to CT antigens that potentially can be up-regulated in clinical settings by chromatin remodeling agents. However, unlike MHC-deficient K562-GM, H1299 cells exhibit class I as well as class II HLA expression; consequently, H1299 cells may induce more allo-reactivity than K562-GM cells when used as whole cell vaccines. In laboratory experiments, we have observed that relative to conventional RIPA-buffer lysates, freeze-thaw lysates of H1299 cells have significantly lower levels of HLA proteins, while retaining high levels of CT-X antigen. In a trial commencing in September 2014 (14-C-0053), thoracic oncology patients who are rendered NED by conventional therapy, yet are at high risk for recurrence, will be randomized to receive H1299 lysates with Iscomatix with or without metronomic cyclophosphamide/celecoxib. Vaccines will be administered monthly for 6 months followed by treatment evaluation one month later. A variety of biologic and immunologic endpoints will be assessed during this First-in-Humans vaccine trial.

在过去的十年中，在一系列临床方案中对100多名胸腔恶性肿瘤患者进行了治疗，这些临床方案检查了DNA脱甲基化剂（Decitabine; DAC）和HDAC抑制剂（例如Romidepsin（DP）），检查了毒性和临床反应。尽管在大约10％的患者中观察到疾病的稳定延长（4-12个月），但这些试验总共表现出没有客观的临床回归。在所有接受DAC输注的患者中，将近四分之一的患者表现出肿瘤组织中p16，Mage-3或NY-ESO-1的表达增加。在接受DAC的几名患者中观察到了对NY-ESO-1的血清学反应超过六个月。接受DP输注的患者中约有50％表现出肿瘤内H3AC和P21的升高。此外，在DP输注后，几名患者在肿瘤活检中表现出增强的NY-ESO-1和MAGE-A3的表达。对激光的微阵列分析从接受DAC，DP或顺序DAC/DP输注的患者的治疗前和治疗后活检中捕获的肿瘤细胞表明，从肺癌基因的签名转移到正常呼吸性上皮elia中观察到的肿瘤细胞。这些发现，以及肿瘤组织中CT-X抗原上调上调的观察，为使用表观遗传方案与免疫学干预措施结合使用，以靶向胸腔恶性肿瘤中的CT-X抗原。通常，胸腔恶性肿瘤表现出低水平，局灶性和异质CT-X基因表达，以及对染色质重塑剂的可变反应。由于CT-X抗原似乎在多能肿瘤细胞中优先表达，因此可以想象，自体性表观遗传修饰的肿瘤细胞可能是靶向癌症干细胞的独特的个性化疫苗，从而促进全身转移。最近的临床努力集中在评估原发性胸腔恶性肿瘤患者的表观遗传改性肿瘤细胞，以及胸外恶性肿瘤转移到胸部的患者，这些患者已通过标准治疗赋予了NED。该疫苗是用Iscomatrix施用的，Iscomatrix是一种专有辅助物，已显示出对纯化的CT-X抗原产生有效的免疫反应；此外，口服塞来昔布将用于抑制已显示可减弱癌症患者治疗疫苗的免疫反应的Treg的活性。迄今为止，已有30名患者参加了这项试验。尽管临床前研究证明了这种方法的可行性，但原发性肿瘤细胞系的可靠推导是一个重大挑战，部分原因是切除的恶性肿瘤的组织学。尽管已经从几个个体建立了细胞系，但由于疾病复发，这些患者没有资格进行疫苗接种。为了避免建立原始细胞系的问题，我们已经确定了包括K562红血球血症在内的几个已建立的线条和H1299肺癌细胞，这些肺癌细胞表现出高水平的CT-X基因表达，而无需用作辅助疫苗的药物操纵。在最近的II期试验中，TGIB在TGIB中，有20例患者接受了六次每月免疫，2.5E7-1E8 LIVE LIVE辐照的K562细胞组成表达GM-CSF（K562-GM），并与分配的口服细胞磷酰胺和Celecoxib（celecoxib）（抑制免疫抑制tregs）作为佐剂治疗，在其恶性肿瘤完全切除后。计量化疗的耐受性良好，未观察到与疫苗相关的毒性。虽然几名患者在接种疫苗后表现出对纯化的CT-X抗原的血清学反应性，但没有一个符合免疫反应的标准。在接受治疗期间，几名在接受治疗期间不典型的无疾病间隔的患者在被取消研究后不久就会迅速复发。这些发现增加了我们分析未检测到的对疫苗的反应和/或分析化学疗法的潜在临床益处。目前，与该审判有关的手稿正在准备出版。在最近的努力中，我们使用H1299肺癌细胞开发了一种疫苗。由于X染色体的扩增，H1299细胞表现出比K562-GM更广泛，更高的CT-X基因表达。因此，H1299细胞可能是诱导CT抗原免疫的更有效疫苗，该疫苗可能通过染色质重塑剂在临床环境中可能上调。但是，与缺乏MHC的K562-GM不同，H1299细胞表现出I类以及II类HLA表达。因此，当用作全细胞疫苗时，H1299细胞可能比K562-gm细胞诱导更多的同种反应性。在实验室实验中，我们观察到，相对于常规的Ripa-buffer裂解物，H1299细胞的冻融裂解物的HLA蛋白水平明显较低，同时保留了高水平的CT-X抗原。在2014年9月开始的试验中（14-C-0053），通过常规疗法进行NED但重复的高风险，将随机分配带有ISComatix的H1299裂解物，具有或不具有分配性环磷酰胺/CelecoxIb 。疫苗将在一个月后每月进行6个月，然后进行治疗评估。在这项首次人类疫苗试验中，将评估各种生物学和免疫终点。