IL-15 TRiKES-based specific immunotherapy of TNBC; resistance mechanisms

基于IL-15 TRiKES的TNBC特异性免疫治疗；

• 批准号: 9751815
• 负责人: SOLDANO FERRONE
• 金额: $ 8.01万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751815
• 关键词: Achievement; Animal Model; Apoptotic; Binding; Biological Models; Bispecific Antibodies; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer cell line; Budgets; Cancer Center; Cell Line; Cells; Chondroitin Sulfate Proteoglycan; Clinical; Clinical Trials Design; Combination immunotherapy; Conduct Clinical Trials; Crosslinker; Cytolysis; Development; Disease; Down-Regulation; Effector Cell; Excision; FCGR3B gene; Failure; Frequencies; Future; Goals; Histocompatibility Antigens Class I; Homologous Gene; Human; Hybrids; Hypoxia; Hypoxia Pathway; Immune; Immunotherapeutic agent; Immunotherapy; In Vitro; Infrastructure; Interleukin-15; Link; Malignant Neoplasms; Mediating; Monoclonal Antibodies; Morbidity - disease rate; Mus; Natural Killer Cells; Neoplasm Metastasis; Normal Cell; Normal tissue morphology; PD-1/PD-L1; PDCD1LG1 gene; Pathway interactions; Patients; Pericytes; Phase I Clinical Trials; Play; Population; Predisposition; Primary Neoplasm; Recurrence; Resistance; Role; SHH gene; SLEB2 gene; Synapses; Testing; Therapeutic; Translating; Treatment Efficacy; Tumor Antigens; Tumor Immunity; Woman; Xenograft procedure; angiogenesis; anti-PD-L1; antigen processing; base; cancer stem cell; clinically significant; combinatorial; crosslink; design; effective therapy; immunological synapse; implementation trial; in vivo; inhibitor/antagonist; malignant breast neoplasm; mortality; neoplastic cell; novel; novel strategies; resistance mechanism; side effect; skills; small molecule; success; theories; triple-negative invasive breast carcinoma; trispecific killer engager; tumor; tumor microenvironment

ABSTRACT The need of an effective therapy for metastatic triple negative breast cancer (TNBC) has prompted us to develop a novel combinatorial immunotherapeutic strategy for this very aggressive subtype of breast cancer. In the design of our strategy we have been guided by the realization that to be effective, a therapy has to eradicate both differentiated TNBC cells and TNBC cancer initiating cells (CICs). According to the cancer stem cell theory, CICs play a major role in disease recurrence and metastatic spread, the major causes of patient morbidity and mortality. Furthermore, immunotherapy has to counteract the multiple escape mechanisms utilized by TNBC cells to avoid immune destruction. Therefore, i) to eliminate both differentiated TNBC cells and TNBC CICs, we have selected as a target chondroitin sulphate proteoglycan 4 (CSPG4), since this TA can mediate the immune destruction of both types of targets. In addition, CSPG4 is selectively up-regulated on activated pericytes in the tumor microenvironment. Therefore, CSPG4 immunotargeting inhibits neoangiogenesis and contributes to the elimination of TNBC cells, even those with CSPG4 loss or down regulation. Lastly CSPG4 has a restricted distribution in normal tissues. Therefore, CSPG4 immunotargeting is not expected to cause side effects because of the targeting of normal tissues. ii) To counteract the escape mechanisms caused by abnormalities in HLA class I antigen processing machinery in TNBC cells, we have selected as effectors NK cells which do not require HLA class I antigen expression for recognition of target cells. To activate and expand the NK cell effector population at the level of the immune synapse, we have added IL-15 to the conventional bispecific NK Cell immune engagers (BiKEs) platform and used it to crosslink the scFv fragments derived from the CSPG4-specific mAb 763.74 to a highly modified camelid CD16 (FCγRIII)-specific scFv fragment . The latter binds to NK cells, while the former to TNBC cells. These novel hybrid molecules which are referred to as IL-15 TriKEs enhance the ability of NK cells to kill tumor cells with limited -if any- damage to normal cells. iii) To restore the susceptibility of TNBC cells to immune lysis we combine IL-15 TriKEs with the small molecule LDE225, an inhibitor of the SHH pathway, since the activation of this pathway up-regulates the level of anti-apoptotic molecules. iv) To enhance the anti-tumor activity of NK cells we disrupt the PD-1/PD-L1 axis with an anti-PD-L1 mAb. We will test the hypothesis that NK cells in combination with IL-15 TriKEs, LDE225 and anti-PD-L1 mAb are effective in eradicating both differentiated TNBC cells and TNBC CICs, both in vitro and in vivo. As a result, TNBC disease recurrence and metastatic spread will be suppressed. Because of the limited budget the proposed studies will be performed only with TNBC cell lines. In future studies the potential clinical significance of the results obtained with cell lines will be assessed with patient derived xenografts. This information will then be translated to a clinical setting in a Phase I clinical trial.

摘要 对转移性三阴性乳腺癌（TNBC）有效治疗的需求促使我们 为这种侵袭性很强的乳腺癌亚型开发一种新的组合免疫策略。在 我们的策略设计是基于这样一种认识，即要想有效，治疗必须 根除分化的TNBC细胞和TNBC癌症起始细胞（CIC）。根据癌症干细胞 根据细胞学说，CIC在疾病复发和转移扩散中起主要作用， 发病率和死亡率。此外，免疫疗法必须抵消多种逃避机制， 利用TNBC细胞来避免免疫破坏。因此，i）为了消除两种分化的TNBC细胞， 和TNBC CIC，我们选择硫酸软骨素蛋白聚糖4（CSPG4）作为靶，因为该TA可以 介导两种类型的靶的免疫破坏。此外，CSPG4在细胞周期中选择性上调。 激活肿瘤微环境中的周细胞。因此，CSPG4免疫靶向抑制 新血管生成，并有助于消除TNBC细胞，即使是CSPG4丢失或降低的细胞 调控最后，CSPG4在正常组织中具有受限的分布。因此，CSPG4免疫靶向是 由于靶向正常组织，预期不会引起副作用。（二）防止逃逸 由于TNBC细胞中HLA I类抗原加工机制异常引起的机制，我们 选择不需要HLA I类抗原表达来识别靶的NK细胞作为效应子 细胞为了在免疫突触水平激活和扩增NK细胞效应子群体，我们 将IL-15添加到常规的双特异性NK细胞免疫抑制剂（BiKE）平台，并使用其交联 将来自CSPG4特异性mAb 763.74的scFv片段转化为来自CSPG4特异性mAb 763.74的scFv片段。 高度修饰的骆驼CD16 (FCγ RIII）特异性scFv片段 .后者与NK细胞结合，而前者与TNBC细胞结合。这些新颖 称为IL-15 TriKE的杂合分子增强NK细胞杀伤肿瘤细胞的能力， 有限的-如果有的话-对正常细胞的损害。iii）为了恢复TNBC细胞对免疫裂解的易感性，我们 将联合收割机IL-15 TriKEs与小分子LDE225（SHH通路的抑制剂）结合，因为 该途径上调抗凋亡分子的水平。iv）增强NK的抗肿瘤活性 我们用抗PD-L1 mAb破坏PD-1/PD-L1轴。我们将检验NK细胞在 与IL-15 TriKE、LDE 225和抗PD-L1 mAb的组合有效地根除了两种分化的 TNBC细胞和TNBC CIC，在体外和体内。因此，TNBC疾病复发和转移性肿瘤的发生率很低。 扩散将被抑制。 由于预算有限，将仅使用TNBC细胞系进行拟定研究。今后 研究将在患者中评估细胞系所得结果的潜在临床意义 异种移植物。然后，这些信息将在I期临床试验中转化为临床环境。

项目成果

Enhanced targeting of triple-negative breast carcinoma and malignant melanoma by photochemical internalization of CSPG4-targeting immunotoxins.

• DOI: 10.1039/c7pp00358g
• 发表时间: 2018-05-16
• 期刊: Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology
• 作者: Eng MS;Kaur J;Prasmickaite L;Engesæter BØ;Weyergang A;Skarpen E;Berg K;Rosenblum MG;Mælandsmo GM;Høgset A;Ferrone S;Selbo PK
• 通讯作者: Selbo PK

CD16-158-valine chimeric receptor T cells overcome the resistance of KRAS-mutated colorectal carcinoma cells to cetuximab.

• DOI: 10.1002/ijc.32618
• 发表时间: 2020-05-01
• 期刊: International journal of cancer
• 影响因子: 6.4
• 作者: Arriga R;Caratelli S;Lanzilli G;Ottaviani A;Cenciarelli C;Sconocchia T;Spagnoli GC;Iezzi G;Roselli M;Lauro D;Coppola A;Dotti G;Ferrone S;Sconocchia G
• 通讯作者: Sconocchia G