Abstract B076: ROME, a novel membrane protein in vertebrates, enhances metastatic phenotype of Ewing sarcoma cells
Bibliographic record
Abstract
Abstract In order to discover genes that can accelerate metastasis of Ewing sarcoma (ES), we performed a genome-wide CRISPR/Cas9 transcriptional activation screen in zebrafish using human ES cells as xenografts. We identified the human INAFM2 gene as a strong driver of metastatic activity of ES cells. INAFM2 is the human homolog of the Drosophila InaF gene and there were no publications about the protein product of this gene in any vertebrate animals. We named the new protein ROME (Regulator of metastasis) and characterized its expression and its role in ES metastasis using multiple in vitro and in vivo models. We established that the human ROME protein is indeed expressed at the plasma membrane. We demonstrated that the ROME protein is both phosphorylated and glycosylated. The functional significance of these posttranslational modifications remains to be determined. Lack of ROME expression in zebrafish embryos resulted in severe developmental defects and early mortality. We performed single cell RNA sequencing of zebrafish embryos that were injected with ROME targeting morpholino. RNA sequencing of ES cell lines that were either overexpressing ROME or lack ROME expression (by CRISPR). We determined that ROME expression regulates Ca++, Wnt and MAPK signaling pathways. Overexpression of ROME in four ES cell lines caused increased cell migration and chemotaxis in vitro, and increased intravasation in zebrafish xenografts. ROME knockout in two ES cell lines decreased chemotaxis. Rescue of ROME expression restored the cells’ ability to migrate in CRISPR knockout clones. In vivo studies by tail vein injection of ES cells in immunodeficient mice demonstrated significantly increased gross metastases with ROME overexpression and decreased gross metastases with ROME knockout compared to the respective control groups. Additional ES xenograft studies where the primary tumor was removed by amputating the affected leg, showed that overexpression of ROME results in enhanced lung metastasis. Analysis of patient tumor RNA sequencing data from many different types of human cancers revealed correlations between ROME expression and both an increased metastatic phenotype and poor patient survival. This provides strong evidence for a substantial role of ROME in ES and cancer metastasis in general. In summary, we present the first ever characterization of the ROME protein in zebrafish embryos and ES cancer cell lines. Our data suggest that ROME is a membrane protein that can regulate cell motility and invasion through different signaling pathways, which results in increased metastatic potential. Citation Format: Anna Molotkova, Emre Deniz, Matthew Swift, Eric Glasgow, Jeffrey Toretsky, Aykut Uren. ROME, a novel membrane protein in vertebrates, enhances metastatic phenotype of Ewing sarcoma cells [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pediatric Cancer Research; 2024 Sep 5-8; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl):Abstract nr B076.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.003 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".