Oncology
Abstract
Clinical studies of cancer patients have shown that overexpression or amplification of thymidylate synthase (TS) correlates with a worse clinical outcome. We previously showed that elevated TS exhibits properties of an oncogene and promotes pancreatic neuroendocrine tumors (PanNETs) with a long latency. To study the causal impact of elevated TS levels in PanNETs, we generated a mouse model with elevated human TS (hTS) and conditional inactivation of Men1 gene in pancreatic islet cells (hTS/Men1-/-). We demonstrated that increased hTS expression was associated with earlier tumor onset and accelerated PanNET development as compared to control Men1-/- and Men1+/ΔN3-8 mice. We also observed decrease in overall survival of hTS/Men1+/- and hTS/Men1-/- mice as compared to control mice. We showed that elevated hTS in Men1-deleted tumor cells enhanced cell proliferation, deregulated cell cycle kinetics and was associated with a higher frequency of somatic mutations, DNA damage and genomic instability. In addition, we analyzed survival of 88 PanNET patients and observed that high TS protein expression independently predicted worse clinical outcome. In summary, elevated hTS directly participates in promoting PanNET tumorigenesis with reduced survival in Men1 mutant background. This work will re-focus attention on new strategies to inhibit TS activity for PanNET treatment.
Authors
Vinod Vijayakurup, Kyungah Maeng, Hye Seung Lee, Benjamin S. Meyer, Sandra Burkett, Akbar Nawab, Michael W. Dougherty, Christian Jobin, Iqbal Mahmud, Timothy J. Garrett, Michael Feely, Kyoung Bun Lee, Frederic J. Kaye, Maria V. Guijarro, Maria Zajac-Kaye
Abstract
Collateral lethality occurs when loss of a gene/protein renders cancer cells dependent on its remaining paralog. Combining genome-scale CRISPR/Cas9 loss-of-function screens coupled with RNA-sequencing in over 900 cancer cell lines, we found that cancers of nervous system lineage, including adult and pediatric gliomas and neuroblastomas, required the nuclear kinase Vaccinia-Related Kinase 1 (VRK1) for their survival in vivo. VRK1 dependency was inversely correlated with expression of its paralog VRK2. VRK2 knockout (KO) sensitized cells to VRK1 loss, and conversely, VRK2 overexpression increased cell fitness in the setting of VRK1 loss. DNA methylation of the VRK2 promoter was associated with low VRK2 expression in human neuroblastomas, and adult and pediatric gliomas. Mechanistically, depletion of VRK1 reduced Barrier-to-Autointegration Factor (BAF) phosphorylation during mitosis, resulting in DNA damage and apoptosis. Together, these studies identify VRK1 as a synthetic lethal target in VRK2 promoter-methylated adult and pediatric gliomas and neuroblastomas.
Authors
Jonathan So, Nathaniel W. Mabe, Bernhard Englinger, Kin-Hoe Chow, Sydney M. Moyer, Smitha Yerrum, Maria C. Trissal, Joana Graca Marques, Jason J. Kwon, Brian H. Shim, Sangita Pal, Eshini Panditharatna, Thomas W. Quinn, Daniel A. Schaefer, Daeun Jeong, David L. Mayhew, Justin Hwang, Rameen Beroukhim, Keith L. Ligon, Kimberly Stegmaier, Mariella G. Filbin, William C. Hahn
Abstract
Cellular stress contributes to the capacity of melanoma cells to undergo phenotype switching into highly migratory and drug tolerant dedifferentiated states. Such dedifferentiated melanoma cell states are marked by loss of melanocyte specific gene expression and increase of mesenchymal markers. Two crucial transcription factors, MITF and SOX10, important in melanoma development and progression have been implicated in this process. In this study we describe that loss of MITF is associated with a distinct transcriptional program, MITF promoter hypermethylation and poor patient survival in metastatic melanoma. From a comprehensive collection of melanoma cell lines, we observed that MITF methylated cultures were subdivided in two distinct subtypes. Examining mRNA levels of neural crest associated genes we found that one subtype had lost the expression of several lineage genes including SOX10. Intriguingly, SOX10 loss was associated with SOX10 gene promoter hypermethylation and distinct phenotypic and metastatic properties. Depletion of SOX10 in MITF methylated melanoma cells using CRISPR/Cas9 confirmed these findings. In conclusion, this study describes the significance of melanoma state and the underlying functional properties explaining the aggressiveness of such states.
Authors
Adriana Sanna, Bengt Phung, Shamik Mitra, Martin Lauss, Jiyeon Choi, Tongwu Zhang, Ching-Ni Jenny Njauw, Eugenia Cordero, Katja Harbst, Frida Rosengren, Rita Cabrita, Iva Johansson, Karolin Isaksson, Christian Ingvar, Ana Carneiro, Kevin Brown, Hensin Tsao, My Andersson, Kristian Pietras, Göran Jönsson
Abstract
Macrophages in the tumor microenvironment have a significant impact on tumor progression. Depending on the signaling environment in the tumor, macrophages can either support or constrain tumor progression. It is therefore of therapeutic interest to identify the tumor-derived factors that control macrophage education. With this aim, we correlated the expression of ADAM proteases, which are key mediators of cell-cell signaling, to the expression of pro-tumorigenic macrophage markers in human cancer cohorts. We identified ADAM17, a sheddase upregulated in many cancer types, as a protein of interest. Depletion of ADAM17 in cancer cell lines reduced the expression of several pro-tumorigenic markers in neighboring macrophages in vitro as well as in mouse models. Moreover, ADAM17-/- educated macrophages demonstrated a reduced ability to induce cancer cell invasion. Using mass spectrometry-based proteomics and ELISA, we identified HB-EGF and AREG, shed by ADAM17 in the cancer cells, as the implicated molecular mediators of macrophage education. Additionally, RNA-seq and ELISA experiments revealed that ADAM17-dependent HB-EGF-ligand release induces the expression and secretion of CXCL chemokines in macrophages, which in turn stimulates cancer cell invasion. In conclusion, we provide evidence that ADAM17 mediates a paracrine EGFR-ligand-chemokine feedback loop, whereby cancer cells hijack macrophages to promote tumor progression.
Authors
Sebastian Gnosa, Laia Puig-Blasco, Krzysztof B. Piotrowski, Marie L. Freiberg, Simonas Savickas, Daniel H. Madsen, Ulrich auf dem Keller, Pauliina Kronqvist, Marie Kveiborg
Abstract
Histopathology, the standard method to assess bone marrow in hematologic malignancies such as myeloproliferative neoplasms (MPNs), suffers from notable limitations in both research and clinical settings. Bone marrow biopsies in patients fail to detect disease heterogeneity; may yield a non-diagnostic sample; and cannot be repeated frequently in clinical oncology. Endpoint histopathology precludes monitoring disease progression and response to therapy in the same mouse over time, missing likely variations among mice. To overcome these shortcomings, we used magnetic resonance imaging (MRI) to measure changes in cellularity, macromolecular constituents, and fat versus hematopoietic cells in bone marrow using diffusion-weighted imaging, magnetization transfer, and chemical shift encoded fat imaging. Combining metrics from these imaging parameters revealed dynamic alterations in bone marrow following myeloablative radiation and transplantation. In a mouse MPLW515L bone marrow transplant model of MPN, MRI detected effects of a JAK2 inhibitor, ruxolitinib, within five days of initiating treatment and identified differing kinetics of treatment responses in sub-regions of the tibia. Histopathology validated MRI results for bone marrow composition and heterogeneity. Anatomic MRI scans also showed reductions in spleen volume during treatment. These findings establish an innovative, clinically translatable MRI approach to quantify spatial and temporal changes in bone marrow in MPN.
Authors
Tanner H. Robison, Manisha Solipuram, Kevin Heist, Ghoncheh Amouzandeh, Winston Y. Lee, Brock A. Humphries, Johanna M. Buschhaus, Avinash Bevoor, Anne Zhang, Kathryn E. Luker, Kristen Pettit, Moshe Talpaz, Dariya Malyarenko, Thomas L. Chenevert, Brian D. Ross, Gary D. Luker
Abstract
Identification of molecular subtypes that reflect different prognoses and treatment responses, especially immune checkpoint inhibitors (ICIs) in esophageal squamous cell carcinoma (ESCC), is essential for treatment decisions. We performed targeted sequencing in 201 patients with ESCC to discover genetic subtypes and validate our findings via multiple data sets. We identified 3 driver genes (FCGBP, GRIN2B, and FRY), and recurrent truncating mutations in FRY impaired its tumor-suppressive function and promoted tumor proliferation. A 3-gene mutation signature (FAT1, FAT3, and FRY) recognized a molecular subtype named “FAT/FRY” with frequent Hippo pathway–related mutations. In multiple ESCC cohorts, the patients with the FAT/FRY subtype had poorer prognosis than did patients in the WT group. Transcriptome analysis indicated that the FAT/FRY subtype was characterized by inactivation of the Hippo pathway, hypoxia, chemoresistance, higher infiltration of CD8+ T cells and activated DCs, and a transcriptome similar to that of cancer responders. Furthermore, the 3-gene signature predicted better survival for patients treated with ICIs, partially explained by its positive correlation with the tumor mutation burden and neoantigen burden. The 3-gene signature is a biomarker to recognize the FAT/FRY molecular subtype, evaluate prognosis, and select potential beneficiaries of ICIs in ESCC.
Authors
Zihang Mai, Jianye Yuan, Hong Yang, Shuogui Fang, Xiuying Xie, Xinye Wang, Jiaxin Xie, Jing Wen, Jianhua Fu
Abstract
Development of resistance to chemo- and immuno- therapies often occurs following treatment of melanoma brain metastasis (MBM). In this scenario, astrocytes cooperate towards MBM progression by upregulating secreted-factors, amongst which we found that monocyte chemoattractant protein-1 (MCP-1) and its receptors, CCR2 and CCR4, are overexpressed in activated astrocytes and in brain metastatic melanoma cells compared to primary lesions. We show that melanoma cells alter astrocytes-secretome and evoke MCP-1 expression and secretion, which in turn enhance vascular hyperpermeability and proliferation, migration, and invasion of CCR2-expressing melanoma cells, while inhibiting MCP-1 rescued this phenotype. Pharmacological or molecular inhibition of MCP-1/CCR2 in MBM mouse model activates an anti-tumor immune-mediated response as revealed by the enhanced infiltration of cytotoxic CD8+ T cells, attenuated immunosuppressive phenotype of tumor-associated macrophages, and reduced infiltration of regulatory T cells, leading to inhibition of MBM progression and prolonged survival. In addition, blocking this key target in MBM, improved the therapeutic response of anti-PD-1 immunotherapy, regardless of the tumor mutational load. These results show that the MCP-1/CCR2 axis polarizes the brain microenvironment towards an anti-inflammatory/pro-tumorigenic phenotype, highlighting the therapeutic relevance of this pathway as a potential immune checkpoint in MBM.
Authors
Sabina Pozzi, Anna Scomparin, Dikla Ben-Shushan, Eilam Yeini, Paula Ofek, Alessio D. Nahmad, Shelly Soffer, Ariel Ionescu, Antonella Ruggiero, Adi Barzel, Henry Brem, Thomas M. Hyde, Iris Barshack, Sanju Sinha, Eytan Ruppin, Tomer Weiss, Asaf Madi, Eran Perlson, Inna Slutsky, Helena F. Florindo, Ronit Satchi-Fainaro
Abstract
BACKGROUND. New therapeutic combinations to improve the outcome of ovarian cancer patients are clearly needed. Preclinical studies with ribociclib (LEE-011), a CDK4/6 cell cycle checkpoint inhibitor, demonstrate a synergistic effect with platinum chemotherapy and efficacy as a maintenance therapy after chemotherapy. We tested the safety and initial efficacy of ribociclib in combination with platinum-based chemotherapy in recurrent ovarian cancer. METHODS. This phase I trial combined weekly carboplatin and paclitaxel chemotherapy with ribociclib followed by ribociclib maintenance in patients with recurrent platinum-sensitive ovarian cancer. Primary objectives were safety and maximum tolerated dose (MTD) of ribociclib when given with platinum and taxane chemotherapy. Secondary endpoints were response rate (RR) and progression-free survival (PFS). RESULTS. Thirty-five patients were enrolled. Patients had a mean 2.5 prior lines of chemotherapy, and 51% received prior maintenance therapy with Poly (ADP-ribose) polymerase inhibitors (PARPi) and/or Bevacizumab. The MTD was 400mg. The most common AEs included anemia (82.9%), neutropenia (82.9%), fatigue (82.9%), and nausea (77.1%). Overall RR was 79.3% with a stable disease (SD) rate of 18% resulting in a clinical benefit rate of 96.6%. The PFS was 11.4 months. RR and PFS did not differ based on number of lines of prior chemotherapy or prior maintenance therapy. CONCLUSIONS. This work demonstrates the combination of ribociclib with chemotherapy in ovarian cancer is feasible and safe. With a clinical benefit rate of 97%, this work provides encouraging evidence of clinical efficacy in patients with recurrent platinum-sensitive disease. TRIAL REGISTRATION. ClinicalTrials.gov NCT03056833. FUNDING. This investigator-initiated trial was supported by Novartis who provided drug and funds for trial execution.
Authors
Lan G. Coffman, Taylor J. Orellana, Tianshi Liu, Leonard G. Frisbie, Daniel Normolle, Kent Griffith, Shitanshu Uppal, Karen McLean, Jessica L. Berger, Michelle Boisen, Madeleine Courtney-Brooks, Robert P. Edwards, Jamie Lesnock, Haider Mahdi, Alexander Olawaiye, Paniti Sukumvanich, Sarah E. Taylor, Ronald Buckanovich
Abstract
Immune checkpoint blockade (ICB) therapy has achieved breakthroughs in the treatment of advanced non-small cell lung cancer (NSCLC). Nevertheless, the low response due to immuno-cold tumor microenvironment (TME) largely limits the application of ICB therapy. Based on the glycolytic/cholesterol synthesis axis, a stratification framework for EGFR wild-type NSCLC was developed to summarize the metabolic features of immuno-cold and immuno-hot tumors. The cholesterol subgroup displays the worst prognosis in immuno-cold NSCLC with significant enrichment of the cholesterol gene signature, indicating targeting cholesterol synthesis is essential for the therapy for immuno-cold NSCLC. Statin, the inhibitor for cholesterol synthesis, can suppress the aggressiveness of NSCLC in vitro and in vivo and also drastically reverse immuno-cold to an inflamed phenotype in vivo which exhibited a higher response to ICB therapy. Moreover, both our in-house data and meta-analysis further support that statin can significantly enhance ICB efficacy. In terms of preliminary mechanisms, statin could transcriptionally inhibit PD-L1 expression and induce ferroptosis in NSCLC cells. Overall, we reveal the significance of cholesterol synthesis in NSCLC and demonstrate the improved therapeutic efficacy of ICB in combination with statin. These findings could provide a innovative clinical insight to treat NSCLC patients with immuno-cold tumors.
Authors
Wenjun Mao, Yun Cai, Danrong Chen, Guanyu Jiang, Yongrui Xu, Ruo Chen, Fengxu Wang, Xuehai Wang, Mingfeng Zheng, Xinyuan Zhao, Jie Mei
Abstract
Despite intensive therapy, children with high-risk neuroblastoma are at risk of treatment failure. We applied a multi-omic system approach to evaluate metabolic vulnerabilities in human neuroblastoma. We combined metabolomics, CRISPR screening and transcriptomic data across >700 solid tumor cell lines and identified dihydroorotate dehydrogenase (DHODH), a critical enzyme in pyrimidine synthesis, as a potential treatment target. Of note, DHODH inhibition is currently under clinical investigation in patients with hematologic malignancies. In neuroblastoma, DHODH expression was identified as an independent risk factor for aggressive disease, and high DHODH levels correlated to worse overall and event-free survival. A subset of tumors with the highest DHODH expression was associated with a dismal prognosis, with a 5-year survival of <10%. In xenograft and transgenic neuroblastoma mouse models treated with the DHODH inhibitor brequinar, tumor growth was dramatically reduced, and survival was extended. Furthermore, brequinar treatment was shown to reduce the expression of MYC targets in three different neuroblastoma models in vivo. A combination of brequinar and temozolomide was curative in the majority of transgenic TH-MYCN neuroblastoma mice, indicating a highly active clinical combination therapy. Overall, DHODH inhibition combined with temozolomide has therapeutic potential in neuroblastoma and we propose this combination for clinical testing.
Authors
Thale Kristin Olsen, Cecilia Dyberg, Bethel Tesfai Embaie, Adele M. Alchahin, Jelena Milosevic, Jane Ding, Jörg Otte, Conny Tümmler, Ida Hed Myrberg, Ellen M. Westerhout, Jan Koster, Rogier Versteeg, Han-Fei Ding, Per Kogner, John Inge Johnsen, David B. Sykes, Ninib Baryawno
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