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.

抽象的 需要进行有效的转移性三重阴性乳腺癌（TNBC）的疗法促使我们提出 为这种非常激进的乳腺癌亚型制定一种新型的组合免疫治疗策略。在 我们的战略的设计我们一直在指导下，意识到要有效，疗法必须 根除分化的TNBC细胞和TNBC癌启动细胞（CICS）。根据癌症的茎 细胞理论，CIC在疾病复发和转移性扩散中起主要作用，是患者的主要原因 发病率和死亡率。此外，免疫疗法必须抵消多重逃生机制 由TNBC细胞利用以避免免疫抑制。因此，i）消除两个分化的TNBC细胞 和TNBC CICS，我们已选择为靶硫酸盐硫酸盐蛋白蛋白聚糖4（CSPG4），因为该TA可以 调解两种靶标的免疫限制。此外，CSPG4在 肿瘤微环境中活化的周细胞。因此，CSPG4免疫靶向抑制 新血管生成并有助于消除TNBC细胞，甚至是CSPG4损失或降低的细胞 规定。最后，CSPG4在正常组织中的分布有限。因此，CSPG4免疫靶向是 由于正常组织的靶向，预计不会引起副作用。 ii）抵消逃生 HLA I类抗原加工机械在TNBC细胞中引起的机制，我们有 选择不需要HLA I类抗原表达以识别目标的效果NK细胞 细胞。为了激活和扩展免疫突触水平的NK细胞效应子群体，我们有 将IL-15添加到常规的双特异性NK细胞免疫转发器（自行车）平台中，并将其用于交联 从CSPG4特异性mAb 763.74到A的SCFV片段 高度修改的骆驼CD16 （FcγRIII） - 特异性SCFV片段 。后者与NK细胞结合，而前者与TNBC细胞结合。这些小说 称为IL-15 Trikes的混合分子增强了NK细胞杀死肿瘤细胞的能力 有限 - 如果对正常细胞的任何损害。 iii）恢复TNBC细胞对免疫裂解的敏感性 由于激活的激活 该途径上调了抗凋亡分子的水平。 iv）增强NK的抗肿瘤活性 细胞我们用抗PD-L1 mAb破坏PD-1/PD-L1轴。我们将测试NK细胞中的假设 与IL-15 Trikes，LDE225和抗PD-L1 MAB结合使用，可有效地消除两个分化 TNBC细胞和TNBC CICS，包括体外和体内。结果，TNBC疾病复发和转移性 传播将被抑制。 由于预算有限，因此只有通过TNBC细胞系进行拟议的研究。将来 研究将用患者评估通过细胞系获得的结果的潜在临床意义 派生的异种移植物。然后，此信息将在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