MétaCan
Menu
← Back to cohort
Record W3043616004 · doi:10.1101/2020.07.17.209015

Roles for the long non-coding RNA <i>Pax6os1</i> / <i>PAX6-AS1</i> in pancreatic beta cell identity and function

2020· preprint· en· W3043616004 on OpenAlexaff
Livia López–Noriega, Rebecca Callingham, Aida Martínez-Sánchez, Sameena Nawaz, Grazia Pizza, Nejc Haberman, Nevena Cvetešić, Marie‐Sophie Nguyen‐Tu, Boris Lenhard, Piero Marchetti, Lorenzo Piemonti, Eelco J.P. de Koning, A. M. James Shapiro, Paul Johnson, Isabelle Leclerc, Benoît Hastoy, Benoit R. Gauthier, Timothy J. Pullen, Guy A. Rutter

Bibliographic record

VenuebioRxiv (Cold Spring Harbor Laboratory) · 2020
Typepreprint
Languageen
FieldMedicine
TopicPancreatic function and diabetes
Canadian institutionsUniversity of Alberta
FundersMedical Research CouncilEuropean CommissionDiabetes UKDiabetes Research and Wellness FoundationInnovative Medicines InitiativeWellcome Trust
KeywordsPAX6BiologyPDX1Small hairpin RNAGene silencingGene knockdownRNA interferenceBeta cellMolecular biologyRNAPAX4Gene expressionGeneMessenger RNAIsletTranscription factorGeneticsInsulinEndocrinologyHomeobox

Abstract

fetched live from OpenAlex

Abstract Aim/Hypothesis Long non-coding RNAs (lncRNAs) are emerging as crucial regulators of beta cell development and function. Here, we investigate roles for an antisense lncRNA expressed from the Pax6 locus (annotated as Pax6os1 in mice and PAX6-AS1 in humans) in beta cell identity and functionality. Methods Pax6os1 expression was silenced in MIN6 cells using siRNAs and changes in gene expression were determined by RNA sequencing or qRT-PCR. Mice inactivated for Pax6os1 and human PAX6-AS1 -null EndoC-βH1 cells, were generated using CRISPR/Cas9 technology. Human islets were infected with lentiviral vectors bearing a targeted shRNA or PAX6-AS1 , which were used to silence or overexpress, respectively, the lncRNA. RNA sequencing or RT-qPCR were used to measure transcriptomic changes and RNA pulldown in mice and human cells followed by mass spectrometry/western blot were performed to explore RNA protein interactions. Results Pax6os1/PAX6-AS1 expression was upregulated at high glucose concentrations in derived beta cell lines as well as in mouse and human islets, and in pancreatic islets isolated from mice fed a high fat diet (n=6, p=0.003) and patients with type 2 diabetes (n=11-5, p<0.01). Silencing or deletion of Pax6os1 / PAX6-AS1 in MIN6 or EndoC-βH1cells increased the expression of several β-cell signature genes, including PDX1 and INS . Female, but not male, Pax6os1 null mice fed a high fat diet showed slightly enhanced glucose clearance. ShRNA-mediated silencing of PAX6-AS1 in human islets robustly increased INS mRNA, enhanced glucose-stimulated insulin secretion and calcium dynamics, while overexpression of the lncRNA exerted opposing effects. Pax6os1/AS-1 interacted with histones H3 and H4 in mouse and human cells, indicating a possible role for this lncRNA in histone modifications in both species. Conclusions Increased expression of PAX6-AS1 at high glucose levels may impair beta cell functionality and thus contribute to the development of type 2 diabetes. Thus, targeting PAX6-AS1 may provide a promising strategy to enhance insulin secretion and improve glucose homeostasis in this disease. Research in context What is already known about the subject? Long non-coding RNAs (lncRNAs) are crucial components of the pancreatic islet regulome, whose misexpression may contribute to the development of diabetes. What is the key question? Is the lncRNA Pax6os1/PAX6-AS1 involved in beta cell functionality and type 2 diabetes? What are the new findings? The expression of Pax6os1/PAX6-AS1 is upregulated in mice fed a high fat diet and in pancreatic islets from type 2 diabetes donors. Overexpression of PAX6-AS1 in human pancreatic islets reduces insulin expression, glucose stimulated secretion and intracellular calcium dynamics. Silencing PAX6-AS1 in human pancreatic islets upregulates insulin expression, enhances glucose stimulated insulin secretion and increases intracellular calcium dynamics. How may this impact the clinic in the foreseeable future? Understanding the genetic factors induced by high glucose/obesity involved in beta cell dysfunction is crucial for the development of new therapies to treat T2D.

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 imitation

Not 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.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.002
Threshold uncertainty score0.008

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0020.001

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.

Opus teacher head0.019
GPT teacher head0.232
Teacher spread0.213 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

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".

Quick stats

Citations6
Published2020
Admission routes1
Has abstractyes

Explore more

Same venuebioRxiv (Cold Spring Harbor Laboratory)→Same topicPancreatic function and diabetes→French-language works237,207→