bioRxiv Science⌕ Search

bioRxiv · 10.1101/795468

Evolutionary analyses guide selection of model systems to investigate proto-oncogene function in ALK and LTK

Abstract

Model systems to investigate oncogene-driven cancer have played an essential role in the development of therapies for cancer. However, not all systems are appropriate for all therapeutic targets. Knowing where and when proto-oncogenes and their interactors originated in evolutionary history is key to understanding which organisms can serve as models. Here we investigate two tyrosine kinase receptors that underlie tumorigenesis in cancer: anaplastic lymphoma kinase (ALK) and leukocyte tyrosine kinase (LTK). In Drosophila melanogaster, Caenorhabitis elegans, and Homo sapiens, the discovery of putative ligands Jeb, Hen-1, and AUG has the potential to accelerate the development of novel therapeutics. However, homology of these ligands and receptors is unclear. We performed an exhaustive search for their homologs spanning the metazoan tree of life. Jeb and Hen-1 were restricted to species that diverged prior to the origin of all vertebrates. No non-vertebrate species had ligands orthologous to AUG. Instead, an ancestral receptor tyrosine kinase and AUG gene were present in early vertebrates and are still solitary in lamprey; we demonstrate that the early embryonic expression of AUG in lamprey parallels its expression in model mammal systems. The presence of ALK and LTK in jawed vertebrates is an evolutionary innovation, as is a previously unrecognized functional convergence within ALK and LTK occurring between actinopterygians and sarcopterygians. Our results provide the phylogenetic context necessary for the selection of model organisms that will provide informative investigations of the biology of these critically important tyrosine kinase receptors, enabling successful therapeutic development.\n\nSignificanceThe anaplastic lymphoma kinase ALK can be oncogenically altered to become a driver of several malignancies, including non-small-cell lung cancer and anaplastic large-cell lymphomas. The development of therapeutics targeting this gene depend on the discovery of its interacting partner ligands in relevant model organisms. ALK is found across most major animal groups including mammals, fishes, and invertebrates. Correspondly, several candidate ligands for ALK and its duplicate LTK have been advanced by research in model species. However their homology to the human ligands and therefore their potential to guide therapeutic development is unknown. Our comparative evolutionary analysis revealed which model organisms had functional receptor-ligand pairings that are informative regarding the role of these genes in human tumorigenesis.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Wang, Z., Dornburg, A., Wang, J., Elizabeth, M., Lopez-Giraldez, F., Townsend, J. P.. 2019-10-07. Evolutionary analyses guide selection of model systems to investigate proto-oncogene function in ALK and LTK. https://doi.org/10.1101/795468

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Arrest-associated transcriptional programmes outweigh apoptosis-associated programmes in treatment-naive pancreatic ductal adenocarcinoma

Background: Pancreatic ductal adenocarcinoma (PDAC) is driven by mutant KRAS, and RAS inhibitors now extend survival in previously treated metastatic disease. Whether treatment shrinks a tumour or only halts its growth depends partly on whether cancer cells die or stop dividing, yet how untreated PDAC cells are balanced between these programmes is unknown. Aims: We aimed to characterise the balance between apoptosis-associated and arrest-associated transcriptional programmes in treatment-naive PDAC and to examine their relationships with tumour subtype, TGM2 expression, tissue organisation, survival and drug sensitivity. Methods and results: We scored 52,999 malignant cells from 18 untreated patients in the Hwang atlas using a transcriptional contrast between genes associated with apoptosis and genes associated with resistance to apoptosis or cell-cycle arrest. In every patient the contrast was shifted toward the arrest-associated side: using equality between the two sides as the dividing threshold, 86.3% of cells with an informative score fell on that side, although the proportion depends on the threshold chosen. The shift recurred in the independent Steele atlas (median 71.6% per sample, range 37.1% to 90.8%). In the discovery cohort, tumours with higher basal-like scores had fewer arrest-side cells, and TGM2 expression was modestly higher on the arrest-associated side in both cohorts. The contrast did not track cell-line drug sensitivity or survival in bulk tumours. Conclusion: Untreated PDAC cells show a reproducible transcriptional bias toward anti-apoptotic and cell-cycle-arrest programmes. Whether this bias predicts cellular responses to treatment remains to be tested.

cancer biology↗

Quantitative live imaging reveals region-dependent medulloblastoma disseminationin the zebrafish brain

Medulloblastoma is the most common malignant pediatric brain tumor, and relapse is frequently driven by dissemination within the central nervous system. Yet the early dynamics of tumor spread remain difficult to resolve in vivo. Here, we orthotopically transplanted human medulloblastoma cells into larval zebrafish and followed individual xenografts by longitudinal high-resolution confocal imaging. ONS-76 showed particularly robust dissemination, with hindbrain tumors spreading into the developing cerebellum in 81.5% of larvae, more than fourfold the frequency observed for forebrain or midbrain tumors. Disseminating cells extended along brain-surface and leptomeningeal-like regions and frequently associated with cerebral vasculature. Quantitative analysis of tumor area, perimeter, compactness, convex hull area, and fractal dimension captured changes in growth and spatial complexity over time. Dasatinib reduced tumor expansion and dissemination-related features in a concentration-dependent manner, while higher doses revealed host toxicity. These findings establish a powerful in vivo model for resolving medulloblastoma dissemination and therapeutic response.

cancer biology↗

Targeting REV1 Exposes Context-Dependent Therapeutic Responses in Colorectal Cancer

Translesion synthesis (TLS) promotes DNA damage tolerance and contributes to chemotherapy resistance by enabling replication across damaged DNA templates. REV1, a central TLS regulator, is a potential therapeutic target to enhance the efficacy of genotoxic chemotherapy. Here, we investigated the therapeutic consequences of pharmacologic and genetic REV1 targeting in colorectal cancer (CRC) and evaluated the sensitivity of patient-derived CRC organoids to the REV1 inhibitor JH-RE-06. JH-RE-06 enhanced oxaliplatin sensitivity in HCT116 cells and 5-fluorouracil (5-FU) sensitivity in HT-29 cells, whereas reciprocal combinations produced limited sensitization, indicating treatment- and cellular context-dependent responses. siRNA-mediated REV1 depletion similarly enhanced chemotherapy sensitivity, supporting REV1 involvement in these responses. MNNG sensitivity assays and MLH1 expression analyses did not reveal consistent treatment-associated changes in mismatch repair function, while targeted HPRT1 sequencing identified no detectable sequence alterations in the region examined. Chemosensitization was not accompanied by consistent increases in caspase-3/7 activation under the conditions tested. REV1 inhibition also caused LC3-II accumulation and, after prolonged inhibitor exposure, reduced beta-tubulin abundance, indicating additional autophagy-associated and cytoskeletal responses. In two patient-derived CRC organoid models, JH-RE-06 exhibited single-agent growth-inhibitory activity, with mean IC50 values of 2.01 micromolar and 1.4 micromolar, respectively. Together, these findings demonstrate context-dependent chemosensitization and organoid growth inhibition following REV1 targeting, suggesting that its therapeutic consequences may extend beyond canonical TLS-mediated DNA damage tolerance.

cancer biology↗