Abstract 1198: Neuropilin-1 blockade chemosensitizes pancreatic cancer cells via dual inhibition of epithelial-mesenchymal transition and autophagy
Bibliographic record
Abstract
Abstract Introduction: Fibrotic and malignant pancreatic ductal adenocarcinoma (PDAC) tumors are often resistant to chemotherapy, and remain a leading cause of morbidity and mortality. Epithelial–mesenchymal transition (EMT), a process by which epithelial cells lose their cell polarity and adhesion to become migratory and invasive mesenchymal stem cells, contributes to chemoresistance in PDAC. Likewise, PDAC tumors benefit from autophagy, a self-degradation pathway that confers a survival benefit against metabolic stress. Over-activation of EMT and autophagy has been linked to increased drug resistance in PDAC. Both these processes are largely mediated by TGFβ1. Recently, we demonstrated the role of neuropilin-1 (NRP-1) in TGFβ1-dependent endothelial-mesenchymal transition and fibrosis in PDAC. However, the potential mechanisms linking NRP-1 with EMT and autophagy-mediated chemoresistance remain unexplored. We hypothesized that NRP-1 blockade will enhance chemotherapeutic effect, by suppressing EMT and autophagy in PDAC. Methods: Human PDAC cells (BxPC-3) or serially gemcitabine exposed chemoresistant (CR-BxPC-3) cells were transfected with siNRP-1 or scramble using DharmaFECT-2. For EMT and chemotherapy studies, transfected cells were treated with TGFβ1 (5ng/mL, 24h), followed by gemcitabine (1μM) for 48h. RNA (24h) and protein (48h) was isolated using TRIzolTM and RIPA, respectively. EMT markers were evaluated by RT-PCR and immunoblotting/immunostaining. For autophagy studies, cells were serum starved post NRP-1 silencing and protein was isolated. For autophagy inhibition, cells were treated with bafilomycin-A1 (100nM) for 24h post NRP-1 silencing and protein was isolated. Autophagy flux (LC3-II/I) and P62 were measured by immunoblotting/immunostaining. Cell viability was evaluated by MTT assay. Results: SiNRP-1 treatment in BxPC-3 and CR-BxPC-3 cells resulted in NRP-1 knockdown and promoted cell-death after chemotherapy. TGFβ1-stimulated BxPC-3 cells demonstrated a morphological change consistent with EMT that was revoked by siNRP-1. Also, siNRP-1 maintained epithelial cellular morphology via gain of epithelial and loss of mesenchymal markers. Remarkably, CR-BxPC-3 cells exhibited enhanced EMT features. SiNRP-1 inhibited EMT and promoted cell-death by overcoming gemcitabine resistance in CR-BxPC-3 cells, possibly through reduced ERK signaling. Moreover, siNRP-1 inhibited autophagy (decreased LC3-II/I ratio and increased P62 levels) following starvation or chemotherapy stress, and enhanced cell-death after chemotherapy and autophagy inhibition in BxPC-3 cells. Conclusions: Our findings define a previously undetermined role of NRP-1 in regulating autophagy and TGFβ1-induced EMT in PDAC, suggesting that NRP-1 may represent a novel therapeutic target to overcome chemoresistance through reduced EMT and autophagy. Citation Format: Pratiek N. Matkar, Krishna K. Singh, Gerald Prud’homme, David Hedley, Howard Leong-Poi. Neuropilin-1 blockade chemosensitizes pancreatic cancer cells via dual inhibition of epithelial-mesenchymal transition and autophagy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1198. doi:10.1158/1538-7445.AM2017-1198
Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.
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 | 0.001 |
| Insufficient payload (model declined to judge) | 0.003 | 0.000 |
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".