Asian nonsmoking populations have a higher incidence of lung cancer compared with their European counterparts. There is a long-standing hypothesis that the increase of lung cancer in Asian never-smokers is due to environmental factors such as second-hand smoke. We analyzed whole-genome sequencing of 30 Asian lung cancers. Unsupervised clustering of mutational signatures separated the patients into two categories of either all the never-smokers or all the smokers or ex-smokers. In addition, nearly one third of the ex-smokers and smokers classified with the never-smoker-like cluster. The somatic variant profiles of Asian lung cancers were similar to that of European origin with G.C>T.A being predominant in smokers. We found EGFR and TP53 to be the most frequently mutated genes with mutations in 50% and 27% of individuals, respectively. Among the 16 never-smokers, 69% had an EGFR mutation compared with 29% of 14 smokers/ex-smokers. Asian never-smokers had lung cancer signatures distinct from the smoker signature and their mutation profiles were similar to European never-smokers. The profiles of Asian and European smokers are also similar. Taken together, these results suggested that the same mutational mechanisms underlie the etiology for both ethnic groups. Thus, the high incidence of lung cancer in Asian never-smokers seems unlikely to be due to second-hand smoke or other carcinogens that cause oxidative DNA damage, implying that routine EGFR testing is warranted in the Asian population regardless of smoking status. Cancer Res; 74(21);

Patient-derived xenografts (PDX) have emerged as an important translational research tool for understanding tumor biology and enabling drug efficacy testing. They are established by transfer of patient tumor into immune compromised mice with the intent of using them as Avatars; operating under the assumption that they closely resemble patient tumors. In this study, we established 27 PDX from 100 resected gastric cancers and studied their fidelity in histological and molecular subtypes. We show that the established PDX preserved histology and molecular subtypes of parental tumors. However, in depth investigation of the entire cohort revealed that not all histological and molecular subtypes are established. Also, for the established PDX models, genetic changes are selected at early passages and rare subclones can emerge in PDX. This study highlights the importance of considering the molecular and evolutionary characteristics of PDX for a proper use of such models, particularly for Avatar trials.

Gastric cancer is the fourth most common cancer diagnosed and the second most frequent cause of cancer-related death worldwide. Multiple factors can contribute to the development of gastric cancer, including H. pylori infection, dietary behaviour and life style, possibly resulting in distinct cancer subtypes with different drug sensitivity profiles. In the present study we searched for gastric cancer mutation patterns in the dataset of the “The Cancer Genome Atlas” (TCGA) and in our collection of patient derived xenografts (PDX). In a second part, we evaluated gene alteration patterns for their implications for drug sensitivity. In both TCGA and our PDX datasets, Whole Exome Sequencing analyses revealed two subsets of gastric tumors characterized by specific mutation signatures, with different types and numbers of genomic alterations. The first subset (60% and 75% of samples) contained lower levels of mutations and was characterized by increased numbers of large chromosomal rearrangements resulting in gene loss or amplifications. The second subset of tumors (25%-40% of samples) revealed higher levels of mutations that were predominantly nucleic acid substitutions and small indels linked to mismatch repair genes including MLH1 or MSH3 and to high microsatellite instability. In both subsets, the mutation spectrum was dominated by C>T transitions with an increase of small indels in the subset of highly-mutated tumors. At the gene level, the genes which were mutated in our gastric PDX collection overlapped to great extent with the mutations found in TCGA tumors, especially regarding the most frequently mutated genes. In the first subset, high levels of gene amplifications and deletions were found, including growth factor receptor amplifications in EGFR and HER2. Furthermore, the mutation frequency in genes associated with drug resistance such as KRAS was decreased. The tumors with growth factor receptor amplification responded consistently to therapies such as Cetuximab or Trastuzumab. In contrast, an increased frequency of mutations in oncogenes and tumor suppressors, including KRAS (n=5/10), PIK3CA (n=5/10) and PTEN (n=7/10), was found in the second subset. The mutational profile of these tumors suggest the use of compounds targeting downstream molecules, such as PIK3CA, or targeting effectors of DNA repair, such as PARP, for anti-cancer therapy. Of note, no association was found between the mutation groups and sensitivity to chemotherapeutic agents such as 5FU, Cisplatin or Paclitaxel. In conclusion, we identified two subsets of gastric tumors both in the TCGA dataset and in our collection of PDX models, characterized by distinct genomic alteration profiles suggesting different therapeutic approaches. Currently, we are assessing drug sensitivity profiles within the two subsets in our PDX models. Citation Format: Anne-Lise Peille, Swee-Seong Wong, Florian Kiefer, Bruno Zeitouni, Armin Maier, Frederic Foucault, Tim Kees, Vincent Vuaroqueaux, Amit Aggarwal, Christoph Reinhard, Heinz Herbert Fiebig. Whole exome sequencing analyses of gastric cancers reveal two distinct genomic alteration patterns with implications in drug sensitivity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-314. doi:10.1158/1538-7445.AM2014-LB-314

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

The combination of tumor targeted therapeutics with PD-L1 checkpoint blockade is being explored as a method to increase the clinical benefits of immunotherapy, and expand response to additional cancer types. Merestinib (Mer) is a kinase inhibitor targeting several oncokinases1 (including MET, MST1R, AXL, MERTK, and MKNK1/2) that can potentially modulate immune function, angiogenesis, as well as target the tumor 1-5. To determine the combinatorial potential with immunotherapy, the effects of Mer were evaluated in vitro on human T cells, PBMCs and murine tumor lines CT26 colon carcinoma (harbors KRASmt G12D expresses low Met/no p-Met/high Axl/p-Axl) and B16F10 melanoma (expressing high Met/pMet/peIF4E). Additionally, the anti-tumor effect of Mer was tested in vivo on established CT26 and B16F10 tumors compared to MET specific TKIs (savolitinib, PF4217903) alone or in combination with PD-L1 antibody (Ab) blockade. In vitro, Mer showed no significant effects on either T cells or PBMCs, but was able to inhibit downstream signaling in both CT26 and B16F10 showing activity on murine tumor cell lines. In vivo, daily Mer monotherapy (6, 12 or 24 mg/kg) showed significant anti-tumor effect at all doses in both CT26 and B16F10, that was not seen with either savolitinib or PF4217903. Concurrent combination of Mer (12 mg/kg) and anti-PD-L1 Ab (0.5 mg qw) in CT26 was found to have anti-tumor activity that was synergistic as compared to each single agent alone. While the effect of Mer monotherapy was lost when treatment ended, tumors continued to regress in the combination group even upon cessation of therapy. The combination was well tolerated and resulted in 90% complete responders compared to 30% with anti-PD-L1 Ab alone, 35 days after completing dosing. To test the ability to generate immunologic memory, complete responders were re-challenged with CT26 cells on the contralateral side. All mice in the combination group resisted re-challenge, showing that Mer/PD-L1 Ab combination was triggering immunologic memory. Although there was no significant change in intra-tumor immune cell populations between groups, combination therapy showed an enhanced and unique intra-tumor immune activation/inflammation gene expression signature compared to PD-L1 Ab monotherapy. The enhanced immune activation of the combination therapy, leading to synergistic anti-tumor efficacy, demonstrates that merestinib has the potential to augment immunotherapy while targeting the tumor directly. This preclinical data provides the rationale for the clinical investigation of merestinib in combination with checkpoint therapies targeting the PD-L1/PD1 axis (NCT02791334). 1 - Yan et al. Invest New Drugs 2013;31:833-44 2 - Balan et al. J Biol Chem 2015;290:8110-20 3 - Eyob et al. Cancer Discov 2013;3:751-60 4 - Lemke G. CSH Persp Biol 2013;5:a009076 5 - Piccirillo et al. Nat Immunol 2014;15:503-11 Citation Format: Sau-Chi Betty Yan, Victoria L. Peek, Jennifer R. Stephens, Um L. Um, Amaladas Nelusha, Colleen A. Burns, Kelly M. Credille, Thompson N. Doman, Scott W. Eastman, Beverly L. Falcon, Gerald E. Hall, Philip W. Iversen, Bruce W. Konicek, Jason R. Manro, Any T. Pappas, Julie A. Stewart, Michael B. Topper, Swee-Seong Wong, Michael Kalos, Ruslan D. Novosiadly, Richard A. Walgren, David Schaer. Combination of an oncokinase inhibitor merestinib with anti-PD-L1 results in enhanced immune mediated antitumor activity in CT26 murine tumor model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5590. doi:10.1158/1538-7445.AM2017-5590