Form 3
Report for Joint/Usage Research Program for Endocrine/Metabolism(Fiscal Year 2022)

Date：(Year)/(Month)/(Day)To Director of Institute for Molecular and Cellular Regulation, Gunma University
Principal Applicant
Institution

Institute of Molecular and Cell Biology (IMCB), A*STAR

Position

PI; Assistant Professor

Name

Adrian Teo

We report on the results of joint research in fiscal 2022 as below.
（Program No. 22-3-1 ）１．Research Title

Protein translation in human insulin gene mutated beta cells

２．Purpose andSignificance of theresearch projectIn this proposal, we focus on the effects of heterozygous human INS gene mutations on protein translational status in both beta cell lines and human induced
pluripotent stem cells (hiPSC)-derived beta-like cells.

３．Period of The Program

April 1, 2022 ～ March 31, 2023

４．Project Members
NameAgeSex

Affiliation

39

M

Institute of Molecular andCell Biology (IMCB),A*STAR, Singapore

24

F

IMCB

PositionRole（Principal Applicant）

Adrian Teo

Position：PI
Degree：Ph.D.
Acquisition date：2011.3

Project director

Research officer

Cell Analysis（Research Collaborators）

Carmen Ching※If additional space is required, please attach a separate sheet.
５．Collaborating Researcher Name ofDiabetes andof IMCR
Laboratory
Metabolic Disorders

1 / 2NameJunShirakawa

Institute for Molecular andCellular Regulation IMCR
Gunma University

６． Research Plans
The insulin gene is translated as preproinsulin, consisting of the signal peptide, B-chain, A-chain, andC-peptide. The signal peptide is cleaved off in the endoplasmic reticulum (ER), forming proinsulin. Mutantproinsulin exhibits a dominant-negative effect by forming a complex with wild-type (WT) proinsulin, trapping
it and decreasing insulin production. Moreover, mutant proinsulin has been shown to decrease beta cellmass by inducing ER stress.
We recently reported that the presence of ER stress, organelle changes and insulin processing defects,
resulting in a decreased amount of insulin secreted but not the ability to secrete insulin. By 9 weeks of expression of mutant human INS, dominant-negative effects of mutant INS were evident and beta cell insulin
secretory capacity declined. INS+/C109Y patient-derived beta-like cells and single-cell RNA-sequencing
analyses then revealed compensatory upregulation in genes involved in insulin secretion, processing andinflammatory response (Diabetologia, 2021).
However, we still do not know the translational status in INS C109Y or G32V mutant beta cells. In thiscollaborative project, we will access the protein translational status in INS C109Y or G32V mutant beta
cells by using in both beta cell lines and human induced pluripotent stem cells (hiPSC)-derived beta-likecells. We also plan to identify the target genes for translational changes by RNA-Seq of polysome fractions.
７．Research results:
Please describe the details of the contribution of the joint research with IMCR in obtaining the results.
1. Protein translation analysis in INS mutant beta cells.
Polysome profiling analysis was conducted in MIN6 cells stably transfected with WT and mutant humanpreproinsulin or INS C109Y mutant hiPSCs-derived pancreatic beta-like cells. And some translationalchanges were detected.
2. Identification of translational target genes by polysome profiling
We isolated polysome fraction in WT and mutant human preproinsulin or INS C109Y mutant hiPSCs derived pancreatic beta-like cells. We performed qPCR analysis of ribosome-free and polysome-bound RNAs
from INS mutant beta cells and detected some candidate genes.
８．Present status of academic conference presentations and research papers associated with the resultsof the joint research, and exchange of information on the joint research with the collaborating researcher atIMCR.
(As much as possible, please state papers that include the names of the collaborating researcher at IMCR
or papers stating that the research was supported by the Joint Research Program with IMCR.
Regarding papers, please send a PDF file together with the report to the email address of the general affairs section of the Institute.) Office of General Affairs: kk-msomu4@jimu.gunma-u.ac.jp① Please list the publications that include the name of the collaborating researcher from IMCR and send aN/A

reprint of each publication to IMCR.

② Please list the publications that include a description that the research was supported by the Joint
N/A
③
N/A

④

Research Program with IMCR and send a reprint of each publication to IMCR.

List up to 3 conferences (name of conference, date of conference, and title of the presentation).
Exchange of information exchange with collaborating researcher from IMCR (please list main points ofcommunication).

We routinely discuss the experimental results (every month).

2 / 2Institute for Molecular andCellular Regulation IMCR
Gunma University