crizotinib and Colonic-Neoplasms

To describe a clinical approach to and outcomes of IVF in reproductive-aged cancer survivors receiving targeted cancer therapies.. Case report.. Not applicable.. The first case is of a female patient with metastatic lung cancer receiving long-term crizotinib, an anaplastic lymphoma kinase inhibitor. The second case is of a female patient with metastatic colon cancer receiving long-term denosumab, a RANKL antibody. Both patients presented desiring fertility.. In vitro fertilization.. Live birth and embryo banking.. The potential impact of targeted therapy on oocytes and pregnancy was investigated via literature review and pharmaceutical company inquiries. After oncologic, fertility, and psychological counseling, both survivors underwent ovarian stimulation, IVF, and preimplantation genetic screening. One couple achieved live births of dizygotic twins via gestational surrogacy. The second couple froze one euploid blastocyst for future fertility. Both survivors are stable from their cancer standpoints.. Successful fertility treatments are possible in the context of exposure to crizotinib and denosumab.

Topics: Adult; Anaplastic Lymphoma Kinase; Antineoplastic Agents; Blastocyst; Carcinoma, Non-Small-Cell Lung; Colonic Neoplasms; Crizotinib; Cryopreservation; Denosumab; Drug Administration Schedule; Female; Fertility Preservation; Fertilization in Vitro; Humans; Infertility, Female; Live Birth; Lung Neoplasms; Molecular Targeted Therapy; Ovulation Induction; Pregnancy; Protein Kinase Inhibitors; Pyrazoles; Pyridines; RANK Ligand; Receptor Protein-Tyrosine Kinases; Surrogate Mothers; Twins, Dizygotic

Companion diagnostics (CoDx) will likely continue to rapidly increase in number and application to disease areas including solid tumors, for example EGFR for gefitinib and ALK fusion gene for crizotinib in non-small-cell lung cancer; KRAS against the use of cetuximab and panitumumab in colorectal cancer; HER2 for trastuzumab in breast cancer. CoDx are an indispensable part of personalized medicine and pharmacogenomics. In CoDx development, there are still many challenges, such as the business model promoting cooperation between diagnostics and pharmaceutical companies, and also the regulations related to CoDx. The FDA notice on the development of CoDx in 2011 recommended the co-development of a new drug and CoDx as the best practice, and the Ministry of Health, Labour and Welfare in Japan also issued a statement in 2013. In addition, the recent discovery of many novel variants in the DNA sequence, advances in sequencing and genomic technology, and improved analytic methods have enabled the impact of germline and somatic mutations to be determined using multiplex diagnosis. The complex challenges to develop CoDx necessitate a close collaboration among academic institutions, regulatory authorities, and pharmaceutical companies. [Review].

Topics: Anaplastic Lymphoma Kinase; Biomarkers, Tumor; Breast Neoplasms; Cetuximab; Colonic Neoplasms; Crizotinib; Diagnostic Uses of Chemicals; Drug Discovery; ErbB Receptors; Gefitinib; Gene Fusion; Guidelines as Topic; Humans; Lung Neoplasms; Molecular Diagnostic Techniques; Molecular Targeted Therapy; Pharmacogenetics; Precision Medicine; Proto-Oncogene Proteins p21(ras); Pyrazoles; Pyridines; Quinazolines; Receptor Protein-Tyrosine Kinases; Receptor, ErbB-2; Trastuzumab

Epithelial cell adhesion molecule (EpCAM) is known to highly expression and promotes cancer progression in many cancer types, including colorectal cancer. While metastasis is one of the main causes of cancer treatment failure, the involvement of EpCAM signaling in metastatic processes is unclear. We propose the potential crosstalk of EpCAM signaling with the HGFR signaling in order to govern metastatic activity in colorectal cancer.. Immunoprecipitation (IP), enzyme-linked immunosorbent assay (ELISA), and fluorescence resonance energy transfer (FRET) was conducted to explore the extracellular domain of EpCAM (EpEX) and HGFR interaction. Western blotting was taken to determine the expression of proteins in colorectal cancer (CRC) cell lines. The functions of EpEX in CRC were investigated by proliferation, migration, and invasion analysis. The combined therapy was validated via a tail vein injection method for the metastasis and orthotopic colon cancer models.. This study demonstrates that the EpEX binds to HGFR and induces downstream signaling in colon cancer cells. Moreover, EpEX and HGF cooperatively mediate HGFR signaling. Furthermore, EpEX enhances the epithelial-to-mesenchymal transition and metastatic potential of colon cancer cells by activating ERK and FAK-AKT signaling pathways, and it further stabilizes active β-catenin and Snail proteins by decreasing GSK3β activity. Finally, we show that the combined treatment of an anti-EpCAM neutralizing antibody (EpAb2-6) and an HGFR inhibitor (crizotinib) significantly inhibits tumor progression and prolongs survival in metastatic and orthotopic animal models of colon cancer.. Our findings illuminate the molecular mechanisms underlying EpCAM signaling promotion of colon cancer metastasis, further suggesting that the combination of EpAb2-6 and crizotinib may be an effective strategy for treating cancer patients with high EpCAM expression.

Topics: Animals; Cell Line, Tumor; Cell Movement; Colonic Neoplasms; Crizotinib; Epithelial Cell Adhesion Molecule; Epithelial-Mesenchymal Transition; Signal Transduction

Three-dimensional (3D) culture systems like tumor spheroids represent useful in vitro models for drug screening and more broadly for cancer biology research, but the generation of uniform populations of spheroids remains challenging. The possibility to properly characterize spheroid properties would increase the reliability of these models. To address this issue different analysis were combined: i) a new device and relative analytical method for the accurate, simultaneous, and rapid measurement of mass density, weight, and size of spheroids, ii) confocal imaging, and iii) protein quantification, in a clinically relevant 3D model. The LoVo colon cancer cell line forming spheroids, treated with crizotinib (CZB) an ATP-competitive small-molecule inhibitor of the receptor tyrosine kinases, was employed to study and assess the correlation between biophysical and morphological parameters in both live and fixed cells. The new fluidic-based measurements allowed a robust phenotypical characterization of the spheroids structure, offering insights on the spheroids bulk and an accurate measurement of the tumor density. This analysis helps overcome the technical limits of the imaging that hardly penetrates the thickness of 3D structures. Accordingly, we were able to document that CZB treatment has an impact on mass density, which represents a key marker characterizing cancer cell treatment. Spheroid culture is the ultimate technology in drug discovery and the adoption of such precise measurement of the tumor characteristics can represent a key step forward for the accurate testing of treatment's potential in 3D in vitro models.

Topics: Antineoplastic Agents; Cell Culture Techniques; Cell Survival; Colonic Neoplasms; Crizotinib; Humans; Spheroids, Cellular; Tumor Cells, Cultured

Small-molecule drugs may complement antibody-based therapies in an immune-oncology setting, yet systematic methods for the identification and characterization of the immunomodulatory properties of these entities are lacking. We surveyed the immumomodulatory potential of 1,402 small chemical molecules, as defined by their ability to alter the cell-cell interactions among peripheral mononuclear leukocytes ex vivo, using automated microscopy and population-wide single-cell image analysis. Unexpectedly, ∼10% of the agents tested affected these cell-cell interactions differentially. The results accurately recapitulated known immunomodulatory drug classes and revealed several clinically approved drugs that unexpectedly harbor the ability to modulate the immune system, which could potentially contribute to their physiological mechanism of action. For instance, the kinase inhibitor crizotinib promoted T cell interactions with monocytes, as well as with cancer cells, through inhibition of the receptor tyrosine kinase MSTR1 and subsequent upregulation of the expression of major histocompatibility complex molecules. The approach offers an attractive platform for the personalized identification and characterization of immunomodulatory therapeutics.

Topics: Cardiac Myosins; Cell Line, Tumor; Colonic Neoplasms; Crizotinib; Humans; Immunomodulation; Myosin Heavy Chains; Pyrazoles; Pyridines; Small Molecule Libraries

The identification and clinical validation of cancer driver genes are essential to accelerate the translational transition of cancer genomics, as well as to find clinically confident targets for the therapeutic intervention of cancers. Here we identified recurrent LMNA-NTRK1 and TPM3-NTRK1 fusions in Korean patients with colon cancer (3 out of 147, 2%) through next-generation RNA sequencing (RNA-seq). NTRK1 fusions were mutually exclusive oncogenic drivers of colon cancer that were accompanied with in vitro potential of colony formation and in vivo tumorigenicity comparable to KM12, a human colon cancer cell line harboring TPM3-NTRK1 fusion. NTRK1-encoded TrkA protein was prevalent in 11 out of 216 Korean (5.1%) and 28 out of 472 Chinese patients (5.9%) from independent cohorts, respectively. The expression level of TrkA was significantly correlated with NTRK1 fusion (p = 0.0192), which was verified by a fluorescence in situ hybridization (FISH). Korean patients with TrkA-positive colon cancer had a marginal but significant shorter overall survival time than TrkA-negative colon cancer [hazard ratio (HR) = 0.5346, 95% confidential interval (CI) = 0.2548-0.9722, p = 0.0411]. In addition, KM12 cell line was sensitive to selective TrkA inhibitors. These results demonstrate that NTRK1 fusion is granted as a clinically relevant target for therapeutic intervention of colon cancer.

Topics: Aged; Animals; Carbazoles; Carcinogenesis; Case-Control Studies; Colonic Neoplasms; Crizotinib; Female; Follow-Up Studies; Furans; Gene Fusion; High-Throughput Nucleotide Sequencing; Humans; Immunoenzyme Techniques; In Situ Hybridization, Fluorescence; Lamin Type A; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Staging; Oncogene Proteins, Fusion; Prognosis; Protein Kinase Inhibitors; Pyrazoles; Pyridines; Real-Time Polymerase Chain Reaction; Receptor, trkA; Republic of Korea; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Survival Rate; Tropomyosin; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Activated RAS GTPase signalling is a critical driver of oncogenic transformation and malignant disease. Cellular models of RAS-dependent cancers have been used to identify experimental small molecules, such as SCH51344, but their molecular mechanism of action remains generally unknown. Here, using a chemical proteomic approach, we identify the target of SCH51344 as the human mutT homologue MTH1 (also known as NUDT1), a nucleotide pool sanitizing enzyme. Loss-of-function of MTH1 impaired growth of KRAS tumour cells, whereas MTH1 overexpression mitigated sensitivity towards SCH51344. Searching for more drug-like inhibitors, we identified the kinase inhibitor crizotinib as a nanomolar suppressor of MTH1 activity. Surprisingly, the clinically used (R)-enantiomer of the drug was inactive, whereas the (S)-enantiomer selectively inhibited MTH1 catalytic activity. Enzymatic assays, chemical proteomic profiling, kinome-wide activity surveys and MTH1 co-crystal structures of both enantiomers provide a rationale for this remarkable stereospecificity. Disruption of nucleotide pool homeostasis via MTH1 inhibition by (S)-crizotinib induced an increase in DNA single-strand breaks, activated DNA repair in human colon carcinoma cells, and effectively suppressed tumour growth in animal models. Our results propose (S)-crizotinib as an attractive chemical entity for further pre-clinical evaluation, and small-molecule inhibitors of MTH1 in general as a promising novel class of anticancer agents.

Topics: Aminoquinolines; Animals; Antineoplastic Agents; Colonic Neoplasms; Crizotinib; Crystallization; Disease Models, Animal; DNA Breaks, Single-Stranded; DNA Repair; DNA Repair Enzymes; Female; Homeostasis; Humans; Mice; Mice, SCID; Models, Molecular; Nucleotides; Phosphoric Monoester Hydrolases; Protein Conformation; Protein Kinase Inhibitors; Proteomics; Proto-Oncogene Proteins; Proto-Oncogene Proteins p21(ras); Pyrazoles; Pyridines; ras Proteins; Substrate Specificity; Xenograft Model Antitumor Assays

Oncogenic alterations in MET or anaplastic lymphoma kinase (ALK) have been identified in a variety of human cancers. Crizotinib (PF02341066) is a dual MET and ALK inhibitor and approved for the treatment of a subset of non-small cell lung carcinoma and in clinical development for other malignancies. Crizotinib can induce apoptosis in cancer cells, whereas the underlying mechanisms are not well understood. In this study, we found that crizotinib induces apoptosis in colon cancer cells through the BH3-only protein PUMA. In cells with wild-type p53, crizotinib induces rapid induction of PUMA and Bim accompanied by p53 stabilization and DNA damage response. The induction of PUMA and Bim is mediated largely by p53, and deficiency in PUMA or p53, but not Bim, blocks crizotinib-induced apoptosis. Interestingly, MET knockdown led to selective induction of PUMA, but not Bim or p53. Crizotinib also induced PUMA-dependent apoptosis in p53-deficient colon cancer cells and synergized with gefitinib or sorafenib to induce marked apoptosis via PUMA in colon cancer cells. Furthermore, PUMA deficiency suppressed apoptosis and therapeutic responses to crizotinib in xenograft models. These results establish a critical role of PUMA in mediating apoptotic responses of colon cancer cells to crizotinib and suggest that mechanisms of oncogenic addiction to MET/ALK-mediated survival may be cell type-specific. These findings have important implications for future clinical development of crizotinib.

Topics: Animals; Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; Cell Line, Tumor; Colonic Neoplasms; Crizotinib; Drug Synergism; Female; Gefitinib; Gene Expression Regulation, Neoplastic; Humans; Mice; Niacinamide; Phenylurea Compounds; Protein Kinase Inhibitors; Proto-Oncogene Proteins; Pyrazoles; Pyridines; Quinazolines; Sorafenib; Tumor Suppressor Protein p53; Xenograft Model Antitumor Assays