We have developed several GEM and GEM-derived allograft (GDA) cancer models for preclinical biomarker and therapeutic development. In addition, responding to growing preclinical needs and supporting collaborative activities initiated by CCR clinical investigators, CAPR specialists also recently embarked on a mission to establish several patient-derived xenograft models (PDX), to be employed for therapeutic discovery in oligo-proliferative lung cancer cases. On the path of building a state-of-art portfolio of preclinical models, CAPR major accomplishments include: 1. Generation of multiple driver-specific GEM and GDA metastatic human-relevant SEOC models along with multiple cell cultures for each Inducible models that have been shown to develop SEOC that resembles the human disease in both molecular and biological properties. Models that retain wild type BRCA1 and BRCA2 and models that are deficient in BRCA1 or 2 were established. Models were validated to respond to PARP inhibition with outcomes similar to those in clinical cases and are currently employed in several collaborative studies (also in collaboration with major Pharma company) pursuing validation of combinatorial strategies to attack SEOC malignancy. In addition, a collection of novel ovarian models is being assembled in which oviduct epithelial cells are genetically programmed to give rise to SEOC carcinogenesis representing a current paradigm that the secretory cells of human fallopian tubes represent the cell of origin for SEOC cancer. 2. Generation of human-relevant GEM and GDA primary GBM models and cell lines. Tractable intracranial transplant GBM models were developed and robust therapeutic evaluation workflows were established. The combination of MEK and PI3 kinase inhibition has been demonstrated to inhibit tumor growth and extend survival, whereas inhibition of either alone proved less effective. Development of a human EGFR-driven erlotinib resistant focally evolving lung adenocarcinoma (LA) model All previously established models of this type develop diffuse adenocarcinoma lesions, making it difficult to follow disease for enrollment into drug studies and to monitor and quantify efficacy. We developed a model wherein the erlotinib resistant human EGFRL858R;T790M drives focal development of LA. The model has been validated for increased accuracy of tumor development and efficacy evaluation. 3. Generation/characterization of GEM-Lung adenocarcinoma models driven by 3 distinct human EML4/ALK fusion variants. Transgenic mice harboring human EMK4/Alk fusion genes representing the most frequent human translocations and crisotinib-resistant forms were generated. Mice were shown to express these proteins specifically in lung leading to the development of LA. Generation/characterization of metastatic GDA-PDAC model and cell cultures. 4. Multiple early passage cell cultures were established from pancreatic cancerous lesions that developed in the KPC GEM model (established by Drs. Tuveson and Hingorani). Such cells transplanted orthotopically into the pancreas of recipient immunocompetent mice develop metastatic pancreatic adenocarcinomas with complete set of characteristics typical for the parental tumor. These models have the highest metastatic rate of any GEM PDAC model reported thus far. 5. We completed the construction and extensively validated three novel GEM strains for the most common p53 missense mutations identified in human cancer cases (R172H, R270H, and R270C). Pancreatic Ductal Adenocarcinoma (PDAC) models harboring these p53 alleles have been generated by inter-crossing these alleles with mice harboring the inducible RasG12D allele and PDX-Cre activating transgene. Mice harboring the common p53-172 mutant have been characterized and shown to develop PDAC with properties similar to the KPC model. 6. As an exploratory technology adoption effort, CRISPR/Cas9 system was optimized to introduce multiple oncogenic mutation in lung and brain cells using in vivo infection by recombinant Adeno-Associated Viral (rAAV) Vectors. Intra-tracheal infusion of rAAV reagents engineered to introduce a common spectrum of pro-oncogenic mutations detected in clinical non-small cell lung cancer cases (s.u. p53, Lkb1, and PTEN deletions, Kras-G12D, -D12V, G13D missense mutations, etc.) led to development of pulmonary adenocarcinomas with 5-7-month latency. This technology is being optimized to enable establishing cohorts of experimental animals using such somatic genome editing strategy to introduce oncogenic events in adult tissues of interest, including (but not limited to) brain, lung and pancreas. 7. Via homologous recombination in ES cells, CAPR model development experts have constructed and validated a melanocyte-specific CreERT2 expressing tamoxifen-inducible 'deletor' mouse strain suitable for preparation of preclinical models for melanomagenesis. Although still considered 'work in progress', it has been confirmed that the inter-breeding of this mouse line with conditional BrafV600E and PTENflox/flox mutations leads, upon topic induction by tamoxifen skin painting, to robust carcinogenesis and formation of cutaneous tumors featuring the melanoma component with high penetrance. 8. In addition to the aforementioned achievements in model development, two integrated databases were developed tor managing (1) efficient GEM breeding and maintenance workflows, and (2) preclinical therapeutic evaluation, workflow execution, and data collection in genetic model for pancreatic cancer.
