Signal Transduction Events and the Regulation of Cell Growth

于2023财政年度，我们的主要活动为与临床研究者合作开发适合其临床试验的新药效学（PD）分析，并于同期开放的试验中实施该等PD分析。我们的具体目标包括：1.确定治疗是否达到患者体内的目标。2.研究治疗对宿主的影响，包括在全身水平以及肿瘤和肿瘤微环境中。3.引进新技术，包括在校内和校外转让我们的技术。4.确定新的药物靶点和机制。于二零二三财政年度，我们合作进行超过60项临床试验。我们在生物标志物分析方面的重点主要集中在三个主要领域：1。免疫PD。2.罕见细胞非免疫性PD（例如，循环肿瘤细胞和循环癌症相关成纤维细胞）。3.基因表达分析，适用于新鲜、冷冻或福尔马林固定、石蜡包埋组织（FFPE）。此外，我们还研究了免疫和非免疫靶向治疗对外周血免疫基因表达的全身影响。对于我们合作的大多数临床试验，我们包括通过多参数流式细胞术评估的免疫PD。我们在多项临床试验中发现并发表了与生存率的相关性。这些探索性数据为了解治疗对外周免疫的影响提供了见解。此外，这些数据还可以作为检查点阻断反应的潜在血液指标，正如我们在2019年与Karzai，Madan，Gulley和Dahut博士领导的临床团队合作发表的论文中所证明的那样，该论文研究了用抗PD-L1抗体durvalumab联合PARP抑制剂olaparib治疗的去势抵抗性前列腺癌患者的免疫特征，揭示了CD 8 + T细胞再生的证据。关于免疫表型研究，我们关注多个群体和亚群，包括单核细胞、肿瘤相关巨噬细胞、髓源性抑制细胞、活化和调节性T细胞以及功能标志物的表达。2023年，我们完成了接受durvalumab联合olaparib治疗的三阴性乳腺癌患者的免疫分析。这项研究是与Jung-Min Lee博士合作完成的，手稿正在准备出版。此外，我们与Brigitte Widemann博士及其团队合作，对1型神经纤维瘤病（NF 1）患者和接受司美替尼治疗的不可手术丛状神经纤维瘤患者的外周血进行免疫分析。在Widemann博士的另一项试验中，我们分析了司美替尼联合西罗莫司治疗的不可切除或转移性恶性外周神经鞘瘤患者的免疫亚群。在2022至2023财年期间，我们与Andrea Apolo博士合作，继续确定卡博替尼与检查点抑制剂nivolumab和ipilimumab联合对全身免疫力的影响。于2023财政年度，我们亦继续与Anish托马斯博士合作，进行小细胞肺癌（SCLC）临床试验的免疫分析及循环肿瘤细胞分析。这包括托马斯博士于2022年在《癌细胞》杂志上发表的循环肿瘤细胞结果。此外，我们分析了接受sacituzumab govitecan + berzosertib治疗的SCLC、肺外小细胞神经内分泌癌或对PARP抑制剂耐药的同源重组缺陷型癌症患者的免疫亚群。此外，与Nitin罗珀博士合作，我们研究了有和没有Notch激活药物的SCLC小鼠肿瘤中的免疫亚群变化。对于循环肿瘤细胞，我们一直在利用我们自己的磁珠富集方法进行CTC计数和表型表征。为了进一步推进CTC表征，我们一直在探索富集和分离CTC的方法。目前，我们正在测试两种仪器和一种利用磁珠和柱的方法。与Anish托马斯博士合作，我们在SCLC患者样本中开始了这项工作，以在多细胞水平和单细胞水平上对其进行表征。我们在2023财年开始了三项试验，以评估腹膜癌病、肾上腺皮质癌和胰腺癌FFPE样本的数字空间基因表达谱。

In FY2023, our main activities were working with clinical investigators to develop new pharmacodynamic (PD) assays tailored to their clinical trials and implementing these PD assays in the trials open during the same period. Our specific objectives include: 1. Determining if a therapy hit its target in the patient. 2. Investigating the impact of the therapy on the host, both at the systemic level, and in the tumor and tumor microenvironment. 3. Introducing new technology, including transfer of our technology intramurally and extramurally. 4. Identifying new drug targets and mechanisms. Throughout FY2023, we collaborated on over 60 clinical trials. Our focus in biomarker analysis was primarily on three main areas: 1. Immune PD. 2. Rare cell non-immune PD (e.g., circulating tumor cells and circulating cancer associated fibroblasts). 3. Analyses of gene expression, applicable to fresh, frozen, or formalin-fixed, paraffin-embedded tissue (FFPE). Furthermore, we investigated the systemic effects of immune and non-immune targeted therapy on immune gene expression in peripheral blood. For the majority of the clinical trials on which we collaborate we are including immune PD as assessed by multiparameter flow cytometry. We have found and published correlations with survival in multiple clinical trials. These exploratory data have provided insights into the impact of therapy on peripheral immunity. Additionally, these data serve as a potential blood-based indicator of response to checkpoint blockade, as demonstrated in our 2019 publication in collaboration with a clinical team led by Drs. Karzai, Madan, Gulley, and Dahut, studying immune profiling of castration-resistant prostate cancer patients treated with the anti-PD-L1 antibody durvalumab in combination with the PARP inhibitor olaparib, revealing evidence of CD8+ T cell reinvigoration. Regarding immune phenotyping studies, we focused on multiple populations and subpopulations, including monocytes, tumor-associated macrophages, myeloid-derived suppressor cells, activated and regulatory T-cells, and the expression of functional markers. In 2023 we completed immune profiling of triple-negative breast cancer patients treated with durvalumab plus olaparib. This study was done in collaboration with Dr. Jung-Min Lee and a manuscript is in preparation for publication. Additionally, we collaborated with Dr. Brigitte Widemann and her team on immune analysis of peripheral blood in neurofibromatosis type 1 (NF1) patients and patients with inoperable plexiform neurofibromas treated with selumetinib. In another trial with Dr. Widemann, we analyzed immune subsets in patients with unresectable or metastatic malignant peripheral nerve sheath tumors treated with selumetinib in combination with sirolimus. During the FY2022 to 2023 period, in collaboration with Dr. Andrea Apolo, we continued to define the impact of cabozantinib on systemic immunity in combination with the checkpoint inhibitors nivolumab and ipilimumab. In FY2023, we also continued our collaborations with Dr. Anish Thomas on immune profiling and circulating tumor cell analyses of small cell lung cancer (SCLC) clinical trials. This includes the publication of our circulating tumor cell results by Dr. Thomas in Cancer Cell in 2022. In addition, we analyzed immune subsets in the patients with SCLC, extra-pulmonary small cell neuroendocrine cancer, or homologous recombination-deficient cancers resistant to PARP inhibitors, who were treated with sacituzumab govitecan plus berzosertib. Additionally, in collaboration with Dr. Nitin Roper, we investigated immune subset changes in the tumor of SCLC mice with and without Notch activating drug. For circulating tumor cells, we have been utilizing our own magnetic bead enrichment method for CTC enumeration and phenotypic characterizations. To further advance CTC characterization we have been exploring methods to enrich and isolate CTCs. Currently, we are testing two instruments and a method utilizing magnetic beads and columns. In collaboration with Dr. Anish Thomas, we initiated this effort in SCLC patient samples to characterize them at a multicellular level and single-cell level. We have started three trials in FY23 to assess digital spatial gene expression profiling in FFPE samples in peritoneal carcinomatosis, adrenocortical carcinoma, and pancreatic cancer.