Abstract LB-329: Enhancing the resolution and accelerating the pace of translational fusion characterization in oncology by RNA sequencing
Bibliographic record
Abstract
Abstract Chromosomal rearrangements are common markers of cancer progression across a wide range of cancer types, and therefore, identification of fusion transcripts in cancer biopsies may have potential to provide tumor-specific insight toward diagnosis, prognosis and precision treatment. Currently, routine methods for fusion detection using fluorescent in-situ hybridization (FISH) provide a low-resolution view of the aberrant fusion transcript. We describe an RNA-Seq approach designed to survey cancer fusions in a single assay by selectively enriching the cancer transcriptome using probes that target the coding regions of over 1385 cancer-associated genes. We tested the performance of the 1385 gene, RNA-Seq Pan-Cancer panel on RNA extracted from 47 patient-derived samples from brain, sarcoma and leukemia, including blood, bone marrow, and formalin-fixed paraffin-embedded (FFPE) samples. Each sample harbored at least one orthogonally verified gene fusion transcript, previously confirmed by FISH or Reverse Transcriptase PCR (RT-PCR). RNA-Seq libraries were prepared from 10-100 ng of total RNA from blood or bone marrow and 20-200 ng total RNA from FFPE tissue and subsequently enriched by hybridization to the Pan-Cancer panel. All samples yielded sufficient library and were sequenced with 76 base-pair paired-end reads on an Illumina MiSeq at 8 samples per flow cell (∼3 million reads per sample). Sequencing data was analyzed using RNA-Seq with STAR aligner and Manta fusion caller. Using this capture-based single-assay approach, we successfully detected fusions commonly associated with leukemia (BCR-ABL1, MLL-MLLT3, MLL-AFF1, RUNX1-ETV6, EBF1-PDGFRB, TCF3-PBX1, IKZF1-PAX5), sarcoma (EWSR1-ATF1, EWSR1-FLI1, JAZF1-SUZ12, SS18-SSX, FUS-DDIT3, FUS-KLF17, YWHAE-FAM22B) and brain cancer (KIAA1459-BRAF) consistent with previously confirmed RT-PCR or FISH results. Several examples of previously unknown fusion partners or additional structural information that were not identified from the FISH or RT-PCR testing were also uncovered in this study. These cases are described in detail. In summary, we show that selective enrichment of RNA-Seq libraries with cancer-specific probes enables detection of known and novel fusions across a broad range of cancer pathologies in a single reaction, creating new opportunities for discovery and translational cancer studies. Citation Format: Lisa C. Watson, Stephen M. Gross, Felix Schlesinger, Anthony Mai, Mariko Kellogg, Steve Lee, Claire Attwooll, Monica Brenca, David Swanson, Andrew Wong, Angelo P. Dei Tos, Claudia Haferlach, Torsten Haferlach, Wolfgang Kern, Roberta Maestro, Manja Meggendorfer, Niroshan Nadarajah, Maurizio Polano, Sabrina Rossi, Marta Sbaraglia, George S. Charames, Gary P. Schroth, Grace DeSantis. Enhancing the resolution and accelerating the pace of translational fusion characterization in oncology by RNA sequencing. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr LB-329.
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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 | 0.002 | 0.002 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.001 | 0.001 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.003 | 0.003 |
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".