Immunotherapy of low grade lymphoid malignancies

低度淋巴恶性肿瘤的免疫治疗

基本信息

批准号：
10929129
负责人：
adrian u wiestner
金额：
$ 167.99万
依托单位：
NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
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/10929129
关键词：
Affinity Agreement Allogenic Antibodies Antibody Therapy Antigen Targeting Antigens Autologous B lymphoid malignancy B-Lymphocytes BCL2 gene Binding Bispecific Antibodies Blood Bone Marrow CD19 gene CD8-Positive T-Lymphocytes COVID-19 booster COVID-19 pandemic COVID-19 vaccination COVID-19 vaccine Cell Line Cell surface Cells Cellular Immunity Chronic Lymphocytic Leukemia Clinic Clinical Clinical Trials Complement Complement 3d Complement Activation Conduct Clinical Trials Cytolysis Deposition Dose Epitopes Evaluable Disease FDA Emergency Use Authorization Fc Receptor Frequencies Futility Glycoproteins Hepatitis B Vaccines Human IgG1 Immune response Immunotherapy Impairment In Vitro Injections Innovative Therapy Interruption Intervention Investigation Leukemic Cell Lymphoma MS4A1 gene Malignant lymphoid neoplasm Measures Mediating Medical Modeling Monoclonal Antibodies Monoclonal Antibody CD20 Monoclonal Antibody Therapy Multiple Myeloma Mus Oryctolagus cuniculus Patients Peripheral Blood Mononuclear Cell Progressive Disease ROR1 gene Reaction Recombinants Recommendation Reporting Resistance Role Sampling Secondary Immunization Series Serology Stem cell transplant T cell differentiation T memory cell T-Lymphocyte Testing Time Vaccination Vaccines Variant Work Xenograft Model Xenograft procedure Zoster Vaccine anti-CD20 antibody libraries arm biological adaptation to stress cancer immunotherapy cell killing cell type chemotherapy chimeric antigen receptor T cells chronic lymphocytic leukemia cell clinical development cohort comparison control coronavirus disease cytotoxic cytotoxicity engineered T cells improved in vivo inhibitor insight invention mouse model neoantigens neoplastic cell novel strategies open label overexpression pandemic disease patient derived xenograft model patient response primary endpoint prospective recruit response rituximab sialic acid binding Ig-like lectin success tositumomab tumor growth vaccination strategy vaccine development vaccine response vaccine trial

Affinity Agreement Allogenic Antibodies Antibody Therapy Antigen Targeting Antigens Autologous B lymphoid malignancy B-Lymphocytes BCL2 gene Binding Bispecific Antibodies Blood Bone Marrow CD19 gene CD8-Positive T-Lymphocytes COVID-19 booster COVID-19 pandemic COVID-19 vaccination COVID-19 vaccine Cell Line Cell surface Cells Cellular Immunity Chronic Lymphocytic Leukemia Clinic Clinical Clinical Trials Complement Complement 3d Complement Activation Conduct Clinical Trials Cytolysis Deposition Dose Epitopes Evaluable Disease FDA Emergency Use Authorization Fc Receptor Frequencies Futility Glycoproteins Hepatitis B Vaccines Human IgG1 Immune response Immunotherapy Impairment In Vitro Injections Innovative Therapy Interruption Intervention Investigation Leukemic Cell Lymphoma MS4A1 gene Malignant lymphoid neoplasm Measures Mediating Medical Modeling Monoclonal Antibodies Monoclonal Antibody CD20 Monoclonal Antibody Therapy Multiple Myeloma Mus Oryctolagus cuniculus Patients Peripheral Blood Mononuclear Cell Progressive Disease ROR1 gene Reaction Recombinants Recommendation Reporting Resistance Role Sampling Secondary Immunization Series Serology Stem cell transplant T cell differentiation T memory cell T-Lymphocyte Testing Time Vaccination Vaccines Variant Work Xenograft Model Xenograft procedure Zoster Vaccine anti-CD20 antibody libraries arm biological adaptation to stress cancer immunotherapy cell killing cell type chemotherapy chimeric antigen receptor T cells chronic lymphocytic leukemia cell clinical development cohort comparison control coronavirus disease cytotoxic cytotoxicity engineered T cells improved in vivo inhibitor insight invention mouse model neoantigens neoplastic cell novel strategies open label overexpression pandemic disease patient derived xenograft model patient response primary endpoint prospective recruit response rituximab sialic acid binding Ig-like lectin success tositumomab tumor growth vaccination strategy vaccine development vaccine response vaccine trial

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项目摘要

Monoclonal antibodies (mAbs) combined with chemotherapy improve survival of lymphoma patients. Fc receptor-dependent effector mechanisms achieve tumor cell killing but also induce antigen loss through trogocytosis. Activation of complement can lead to cell lysis and invariably deposits complement C3 activation fragments on the cell surface. For chronic lymphocytic leukemia (CLL) it is well established that not all of the mAb-targeted B cells are killed during these reactions. Instead, substantial numbers of surviving B cells that have lost CD20 due to trogocytosis and are covalently tagged with the complement breakdown product C3d are readily demonstrable in blood and bone marrow. We tested the hypothesis that covalently attached C3d fragments deposited on B cells by anti-CD20 mAbs constitute a neoantigen suitable for targeting antigen loss variants and, when used in combination with a complement fixing mAb, can increase efficacy of mAb therapy. We generated mouse and rabbit mAbs specific for C3d and selected three mAbs that bind distinct epitopes to construct human IgG1 chimeric mAbs. In lymphoma xenografts, co-administration of anti-C3d mAbs with either rituximab or ofatumumab was more effective than the anti-CD20 mAb alone. To extend the application of this invention to Multiple Myeloma (MM) we investigated the combination of the anti-C3d mAbs with the anti-CD38 mAb daratumumab (DARA) that is widely used in patients with MM. In xenograft models, the combination of DARA + anti-C3d antibody was superior to single agent DARA. In conclusion, our findings indicate that C3d-targeting with a specific mAb can provide a decisive second hit to enhance the efficacy of complement fixing mAbs commonly used in lymphoma and MM therapy. In a collaborative agreement with Genmab, we investigated epcoritamab, a CD20xCD3 T cell engaging bispecific antibody in CLL samples in vitro and in the patient-derived xenograft model. Epcoritamab, is in clinical development for B cell malignancies. We found that epcoritamab effectively induced autologous T cells to lyse CLL cells in vitro. Compared to samples from treatment-nave patients, epcoritamab-mediated cytotoxicity was higher in PBMCs from patients being treated with a BTKi, including for patients who had progressive disease on BTKi therapy. Epcoritamab induced CLL cell lysis was correlated with effector to target ratio but not to CD20 expression. Epcoritamab expanded CD4+ and CD8+ T cells, induced memory T-cell differentiation and promoted Th1 polarization; attributes associated with long-term protective responses to cancer immunotherapy. We also tested epcoritamab in our PDX mouse model. Compared to controls treated with non-targeting B12, the median leukemic cell burden was reduced by 71% after one injection of epcoritamab and by 94% after a second injection (P = .003). There was no apparent difference in efficacy of epcoritamab against CLL cells obtained from treatment-nave patients or patients who were progressing but remained on continuous BTKi treatment. Reflecting the situation in patients, the activity of the bsAb in this model depends entirely on effector functions of autologous T cells. The Bcl-2 inhibitor venetoclax, combined with anti-CD20 antibodies, is a commonly used next therapy for patients resistant to or intolerant of BTKis. We tested single agent epcoritamab and venetoclax and their combination in PBMCs from treatment-nave, BTKi-treated, and BTKi resistant patients. In all 3 settings, the combination was significantly more cytotoxic than either agent alone. While most clinically advanced T cell engaging bsAbs for B cell malignancies target CD20, there are many opportunities to develop bsAb against other targets. We previously reported on a bsAb targeting ROR1. More recently, we focused on Siglec-6, a suitable target for antibody-based therapy due to its overexpression on CLL cells and coincidental absence on most healthy cell types. The Siglec-6 binding antibody JML-1 was isolated from a patient cured of CLL by allogeneic stem cell transplantation. CAR-T cells specific for Siglec-6 based on the sequence of JML-1 effectively targeted CLL cells in vitro and in a xenograft mouse model. Using the patients antibody library our collaborator identified additional high affinity clones targeting Siglec-6 and engineered T-cell recruiting bsAbs. In vitro and in vivo these bsAbs lysed Siglec-6 expressing cell lines and primary CLL cells, while sparing most normal B cells. Using the MEC-1 CLL model, we demonstrated that an anti-Siglec-6 in a dual-affinity re-targeting (DART) format inhibited tumor growth and extended survival, comparable to a CD19-targeting control T-bsAb. Patients with CLL have impaired responses to vaccines, and work within our group has yielded insights informing clinical approaches to vaccination in CLL. In two open-label, single-arm clinical trials, we measured the effect of BTKis on de novo immune response against recombinant hepatitis B vaccine (HepB-CpG) and recall response against recombinant zoster vaccine (RZV, Shingrix) in CLL patients who were treatment-naive or on BTKi therapy (ibrutinib or acalabrutinib) for 6 months. The primary endpoint was serologic response to HepB-CpG (anti-HBs 10 mIU/mL) and RZV (4-fold increase in anti-glycoprotein E (gE)). 78 patients received HepB-CpG and 116 patients RZV. Among patients receiving HepG-CpG; 1 (3.8%) of 26 patients on BTKi, and 9 (28.1%) of 32 treatment-nave patients responded (P =.017). At that point, we closed the HepB-CpG arm of the study to patients on BTKi therapy due to futility. In 106 patients evaluable following RZV vaccination, the humoral response rate was significantly higher in the treatment-naive cohort (76.8%) compared to patients receiving a BTKi (40.0%; P = .0002). Similar to humoral responses, the rate of cellular immunity was significantly higher in the treatment-naive cohort (70.0%) compared to patients treated with a BTKi (41.3%; P = .0072). Median anti-gE T-cell frequencies rose from 36 cells per million prior to vaccination to 475 cells per million after completing the vaccine series. There was no difference in T-cell frequencies at baseline between cohorts. Interestingly, 39.0% of subjects attained a cellular immune response in absence of a serologic response. With the onset of the COVID pandemic and the development of vaccines for SARS-CoV2, clinical need for effective vaccination strategies became paramount. Due to the distribution of COVID vaccines under Emergency Use Authorization we could not set up prospective interventional vaccine studies. Instead, we prospectively measured anti-spike (S) Ab titers in our clinic to assess patients response to SARS-CoV2 vaccination. Further, our studies on vaccine responses in years prior to the pandemic inspired our recommendation to interrupt BTKi treatment for up to 3 weeks at the time of booster vaccination. Our strategy was a response to the relatively low success rate of the primary vaccine series in patients with CLL and informed by our investigations on the rate of BTK resynthesis. The recommendation was judiciously shared with patients expected not to suffer negative consequences as projected from our observation that medically indicated, short dosing interruptions do not reduce the long-term benefit of ibrutinib therapy. Indeed, our analysis indicates that patients who interrupted BTKi had higher anti-S Ab titers than those who continued around the time of SARS-CoV2 booster vaccination.

单克隆抗体（mAb）与化学疗法结合改善了淋巴瘤患者的存活。 FC受体依赖性效应子机制可实现肿瘤细胞的杀伤，但也通过曲霉菌诱导抗原丧失。补体的激活会导致细胞裂解，并总是沉积在细胞表面上补充C3激活片段。对于慢性淋巴细胞性白血病（CLL），众所周知，在这些反应中，并非所有受mAb靶向的B细胞都被杀死。取而代之的是，由于Trogococytosis而丢失CD20并与补体分解产物C3D共价标记的大量存活的B细胞在血液和骨髓中很容易证明。我们检验了以下假设：通过抗CD20 mABS沉积在B细胞上的C3D片段构成一种新抗原，适合于靶向抗原损失变异，并且与补体固定MAB结合使用时，可以提高MAB治疗的效率。我们生成了针对C3D的小鼠和兔MABS，并选择了三个MAB，结合不同表位以构建人IgG1嵌合mAb。在淋巴瘤异种移植物中，抗C3D mAb与利妥昔单抗或Ofatumumab的共同给药比单独的抗CD20 MAB更有效。为了扩展本发明的应用到多发性骨髓瘤（MM），我们研究了抗C3D mAb与抗CD38 mAb daratumumumab（dara）的组合，该抗CD38 mAb（dara）广泛用于MM患者。在异种移植模型中，DARA +抗C3D抗体的组合优于单药DARA。总之，我们的发现表明，具有特定MAB的C3D靶向可以提供决定性的第二次命中，以增强在淋巴瘤和MM治疗中常用的补体固定MAB的功效。在与Genmab的一项协议中，我们研究了Epocoritamab，这是CD20XCD3 T细胞在体外和患者衍生的异种移植模型中与CLL样品中的双特异性抗体参与的CD20XCD3 T细胞。 Eponcoritamab，正在用于B细胞恶性肿瘤的临床发育。我们发现，eporitamab有效地诱导自体T细胞在体外裂解CLL细胞。与治疗nave患者的样本相比，在接受BTKI治疗的患者的PBMC中，Epocoritamab介导的细胞毒性较高，包括在BTKI治疗中患有进行性疾病的患者。 Epocoritamab诱导的CLL细胞裂解与效应子与靶比率相关，但与CD20表达不相关。 Eponcoritamab扩展了CD4+和CD8+ T细胞，诱导记忆T细胞分化并促进Th1极化。与对癌症免疫疗法的长期保护反应相关的属性。我们还在PDX小鼠模型中测试了epcoritamab。与用非靶向B12治疗的对照组相比，一次注射Epcoritamab后，白血病细胞负担中位数减少了71％，第二次注射后减少了94％（p = .003）。 eporitamab对从进步但仍处于连续BTKI治疗的患者或患者中获得的CLL细胞的疗效没有明显差异。反映患者的情况，该模型中BSAB的活性完全取决于自体T细胞的效应函数。 Bcl-2抑制剂Venetoclax与抗CD20抗体结合使用，是一种常用的下一个疗法，适用于对BTKIS耐药或不耐受的患者。我们测试了单药epcoritamab和venetoclax及其在治疗范围内的PBMC中的组合，BTKI治疗和BTKI耐药患者。在所有3个设置中，组合的细胞毒性明显比单独的任何一种药物要多得多。虽然大多数临床上先进的T细胞与B细胞恶性肿瘤的靶标CD20吸引BSABS，但有很多机会针对其他靶标开发BSAB。我们以前报告了针对ROR1的BSAB。最近，我们专注于SIGLEC-6，这是基于抗体治疗的合适靶标，这是由于其对CLL细胞的过表达以及对大多数健康细胞类型的偶然缺失。通过同种异体干细胞移植从CLL治疗的患者中分离出SIGLEC-6结合抗体JML-1。基于JML-1的序列在体外和异种移植小鼠模型中有效靶向的CLL细胞的序列对SIGLEC-6的特异性CAR-T细胞。使用患者抗体库，我们的合作者确定了针对SIGLEC-6和设计T细胞招募BSAB的其他高亲和力克隆。在体外和体内，这些BSABS裂解Siglec-6表达细胞系和原代CLL细胞，同时散发大多数正常B细胞。使用MEC-1 CLL模型，我们证明了双亲和力重新靶向（DART）格式中的抗Siglec-6抑制了肿瘤的生长和扩展生存期，可与CD19靶向对照T-bsab相当。 CLL患者对疫苗的反应受损，并且我们小组的工作已经产生了有关CLL临床疫苗接种方法的见解。在两项开放标签的单臂临床试验中，我们测量了BTKI对重组丙型肝炎B疫苗（HEPB-CPG）（HEPB-CPG）的影响，并针对重新组合疫苗（RZV，Shingrix）对BTKIIB或ACARIB的CLL患者（RZV，Shingrix）的召回反应。主要终点是对HEPB-CPG（抗HBS 10 MIU/mL）和RZV（抗糖蛋白E（GE）增加4倍）的血清学反应。 78例患者患有HEPB-CPG和116例RZV患者。在接受HEPG-CPG的患者中；在BTKI的26例患者中，有1名（3.8％），32名治疗nave患者中有9例（28.1％）反应（p = .017）。那时，由于徒劳无益，我们将研究的HEPB-CPG部门关闭给了BTKI治疗的患者。在RZV疫苗接种后可评估的106例患者中，与接受BTKI的患者相比，治疗群体中的体液反应率明显更高（76.8％）（40.0％； P = .0002）。与体液反应相似，与接受BTKI治疗的患者相比，治疗群体中的细胞免疫率明显更高（70.0％）（41.3％； P = .0072）。疫苗接种之前，抗GE T细胞频率从每百万的36个细胞上升到完成疫苗系列后的475个细胞。队列之间基线时T细胞频率没有差异。有趣的是，在没有血清学反应的情况下，有39.0％的受试者获得了细胞免疫反应。随着共证大流行的开始和SARS-COV2疫苗的开发，对有效疫苗接种策略的临床需求变得至关重要。由于在紧急使用授权下分配了共vid疫苗，我们无法进行前瞻性介入疫苗研究。取而代之的是，我们前瞻性测量了诊所中的抗尖峰滴度，以评估患者对SARS-COV2疫苗接种的反应。此外，在大流行之前，我们对疫苗反应的研究启发了我们建议在加强疫苗接种时中断BTKI治疗长达3周的建议。我们的策略是对CLL患者主要疫苗系列成功率相对较低的响应，并通过我们对BTK重新合成速率的研究得出的答复。明智地与预期不会遭受负面后果的患者明智地分享了这一建议，因为我们的观察结果表明，短期剂量中断不会降低ibrutinib疗法的长期益处。实际上，我们的分析表明，中断BTKI的患者的抗S AB滴度比SARS-COV2促进疫苗接种时继续的患者更高。