Reversing Drug Resistance in Tumors with Clickable Antibody Pairs

项目总结/摘要-胃癌和乳腺癌的发病率正在迅速增加， 第四和第五大癌症死亡原因。临床分类为HER 2阳性的患者 （ERBB 2扩增和/或2+/3+蛋白过表达，免疫组织化学法），抗体-药物偶联物 (ADC)延长无进展生存期和总生存期。然而，这些疗法在HER 2-低表达中具有低活性。 癌细胞，并不是所有的HER 2阳性肿瘤受益，甚至那些最初反应不可避免地发展 时间的阻力。根据我们在HER 2异质性患者来源的异种移植物中获得的临床前数据， 证明HER-ADC内吞作用的增加增强了治疗效果，我们开发了 抗体递送方法导致癌细胞中ADC内化增加15.5倍。我们 一种新的方法使用帕妥珠单抗和曲妥珠单抗药物抗体， 结合不同的HER 2结构域，以增加HER 2-ADC复合物的数量，并进一步提高HER 2-ADC复合物的速率。 HER 2-ADC内吞作用。我们已经产生的抗体PET成像研究表明，我们的 一种方法增加了含有HER 2高和HER 2低的异质性肿瘤中89 Zr-ADC的摄取 癌细胞在这里，我们将优化两个表位不同的抗体-ADC生物分子的可点击对，以增强 乳腺癌和胃癌耐药模型中的肿瘤靶向和药物递送。除了测试潜在的 我们的方法在癌细胞系和类器官，我们将进行随机成像和治疗研究 在代表三种肿瘤群体的患者来源的乳腺和胃异种移植物中： HER 2异质性、ADC不合格肿瘤和ADC耐药肿瘤。我们将通过分子成像 （89 Zr-抗体PET）、安全性、药代动力学特征和单独ADC（无点击）与 与点击对缀合的ADC加帕妥珠单抗（点击）。这些随机临床前研究将使我们 以确定分子特征，赋予药物敏感性或耐药性，这一有前途的研究方法。 目标1将优化帕妥珠单抗/ADC点击对，与对照相比， 与ADC单一疗法和Aim 2将验证抗体点击对作为一种新的治疗方法的使用。 这两个目标将为使用抗体点击对增强药物治疗提供重要的新临床前数据。 交付，这可以为许多未来的研究提供良好的基础，包括临床 使用点击对来增强药物递送的翻译以及对其他药物递送的潜在更广泛应用。 膜受体和异质性肿瘤。拟议研究的长期转化目标 是为利用抗体点击来预防或延缓耐药性的临床试验奠定基础， 异质性乳腺癌和胃癌患者。

PROJECT SUMMARY/ABSTRACT – The incidence of gastric and breast cancers is increasing rapidly, rating fourth and fifth leading causes of cancer mortality worldwide. In patients clinically classified as HER2-positive (ERBB2 amplification and/or 2+/3+ protein overexpression by immunohistochemistry), antibody-drug conjugates (ADC) prolong progression-free and overall survival. However, these therapies have low activity in HER2-low cancer cells, and not all HER2-positive tumors benefit, or even those who initially respond inevitably develop resistance over time. Guided by preclinical data we obtained in HER2 heterogeneous patient-derived xenografts demonstrating that an increase in HER-ADC endocytosis enhances therapeutic efficacy, we developed approaches of antibody delivery that result in a 15.5-fold increase of ADC internalization in cancer cells. Our novel approach uses pertuzumab and trastuzumab-drug antibodies that click at the surface of cancer cells upon binding distinct HER2 domains to increase the number of HER2-ADC complexes and further enhance the rate of HER2-ADC endocytosis. Antibody-PET imaging studies that we have generated demonstrate that our approach increases the uptake of 89Zr-ADC in heterogeneous tumors containing HER2-high and HER2-low cancer cells. Here, we will optimize clickable pairs of two epitope-distinct antibody-ADC biomolecules to enhance tumor targeting and drug delivery in resistant models of breast and gastric cancer. In addition to test the potential of our approach in cancer cell lines and organoids, we will perform randomized imaging and therapeutic studies in patient-derived breast and gastric xenografts representing three tumor populations: ADC-eligible tumors of HER2 heterogeneity, ADC-ineligible tumors, and ADC-resistant tumors. We will determine the molecular imaging (89Zr-Antibody PET), safety, pharmacokinetic profile, and therapeutic efficacy of ADC alone (no-click) versus ADC plus pertuzumab conjugated with clicking pairs (click). These randomized preclinical studies will allow us to identify molecular features that confer drug sensitivity or resistance to this promising investigational approach. Aim 1 will optimize pertuzumab/ADC clicking pairs with improved tumor uptake and drug delivery when compared with ADC monotherapies and Aim 2 will validate the use of antibody clicking pairs as a new therapeutic approach. The two aims will provide important new preclinical data on the use of antibody clicking pairs to enhance drug delivery, which could provide an excellent foundation for many future investigations, including the clinical translation of using clicking pairs to enhance drug delivery and the potential broader application to other membrane receptors and heterogeneous tumors. The long-term translational objectives of the studies proposed are to establish a foundation for a clinical trial using antibody clicking to prevent or delay drug resistance in patients with heterogeneous breast and gastric cancers.