News and Publications


Publication

Published:
29 Mar 2022

A single-cell atlas of non-haematopoietic cells in human lymph nodes and lymphoma reveals a landscape of stromal remodelling.

Study was conducted to largely update non-haematopoietic cell (NHC) taxonomy in human lymph nodes (LN) and analysis of disease status. A rich resource and deeper insights into LN and lymphoma biology to advance lymphoma management and therapy is provided.


Nature Cell Biology     Published: March 24, 2022
DOI 10.1038/s41556-022-00866-3

Research Outline (PDF In Japanese language)

Publication

Published:
19 Jan 2022

DAJIN enables multiplex genotyping to simultaneously validate intended and unintended target genome editing outcomes

Genome editing can introduce designed mutations into a target genomic site. Recent research has revealed that it can also induce various unintended events such as structural variations, small indels, and substitutions at, and in some cases, away from the target site. These rearrangements may result in confounding phenotypes in biomedical research samples and cause a concern in clinical or agricultural applications. However, current genotyping methods do not allow a comprehensive analysis of diverse mutations for phasing and mosaic variant detection.


Here, we developed a genotyping method with an on-target site analysis software named Determine Allele mutations and Judge Intended genotype by Nanopore sequencer (DAJIN) that can automatically identify and classify both intended and unintended diverse mutations, including point mutations, deletions, inversions, and cis double knock-in at single-nucleotide resolution. Our approach with DAJIN can handle approximately 100 samples under different editing conditions in a single run. With its high versatility, scalability, and convenience, DAJIN-assisted multiplex genotyping may become a new standard for validating genome editing outcomes.

PLOS Biology     Published: January 18, 2022
DOI 10.1371/journal.pbio.3001507

Research Outline (PDF In Japanese language)

Event

Published:
Posted 27 Jan 2022

20th Anniversary Commemorative Symposium of the University of Tsukuba, Laboratory Animal Research Center (LARC) - (Information in Japanese Only)

REGISTRATION CLOSED *** EVENT COMPLETED ***


→ REGISTRATION CLOSED *** EVENT COMPLETED *** Sympisium Online Event

Symposium Poster Download (Information Available in Japanese Only) PDF (4.1 Mb)


Symposium Program
(Please Note: Symposium will be presented in Japanese language only)

Moderator: TMRC Professor Fumihiro Sugiyama

14:00-14:05
Greetings from the President of University of Tsukuba
President Kyosuke Nagata

14:05-14:10 Guest congratulations
Director, University Research Infrastructure Development Division, Ministry of Education, Culture, Sports, Science and Technology
Mr. Ichiro Kuronuma

14:10-14:30
Outline explanation of Laboratory Animal Resource Center / TMRC, University of Tsukuba
TMRC Chief Satoshi Takahashi


Commemorative Symposium
Chair: Satoshi Takahashi

14:30-15:10
Institute of Medical Science, University of Tokyo and his Stanford University
Professor Hiromitsu Nakauchi
“Creating organs from iPS cells: a cross-border challenge”

15:10-15:50
Keio University School of Medicine
Professor Hideyuki Okano
“Medical research using iPS cells and genetically modified primate technology”

15:50-16:30
Tohoku University School of Medicine and Tohoku Medical Megabank Organization
Professor Masayuki Yamamoto
“Molecular mechanism and pathophysiology of oxidative stress response”

16:30-17:10
University of Tsukuba International Institute for Integrative Sleep Medicine
Professor Masashi Yanagisawa
“Challenge the mystery of sleep -in search of the substance of ‘sleepiness’- “

Closing remarks
TMRC Professor Fumihiro Sugiyama


Award Announcement

Published:
21 Dec 2021

Professor Yamazaki of TMRC selected for 18th Japan Society for the Promotion of Science Award (2021)

Professor Satoshi Yamazaki of TMRC’s Stem Cell Therapy Laboratory was selected to received the “18th Japan Society for the Promotion of Science Award 2021” from JSPS. The award recognised his research work in “Development of Hematopoietic Stem Cell Ex Vivo Expansion”.

Each year, “The Japan Society for the Promotion of Science Award” is presented to high achieving researchers, as encouragement and recognition of their outstanding research creativity and ability at early stages in their careers. Award winners are recognised for their high potential to become Japan’s future leaders, working at the world’s highest levels of academic research and discovery.

→ For more details about the JSPS award, please see the Japan Society for the Promotion of Science web page (external website).

→ The 18th (2021) Japan Society for the Promotion of Science Award winners and reasons for the award (PDF Download - External JSPS Homepage in Japanese language only)


→ See more about TMRC's Stem Cell Therapy Laboratory, led by Professor Yamazaki.

Publication

Published:
11 Nov 2021

The transcriptional corepressor CtBP2 serves as a metabolite sensor orchestrating hepatic glucose and lipid homeostasis

Non-alcoholic fatty liver disease (NAFLD) constitutes a metabolic disorder with high worldwide prevalence and increasing incidence. The inflammatory progressive state, non-alcoholic steatohepatitis (NASH), leads to liver fibrosis and carcinogenesis. Here, we evaluated whether tyrosinase mutation underlies NASH pathophysiology. Tyrosinase point-mutated B6 (Cg)-Tyrc-2J/J mice (B6 albino) and C57BL/6J black mice (B6 black) were fed with high cholesterol diet (HCD) for 10 weeks. Normal diet-fed mice served as controls.


HCD-fed B6 albino exhibited high NASH susceptibility compared to B6 black, a phenotype not previously reported. Liver injury occurred in approximately 50% of B6 albino from one post HCD feeding, with elevated serum alanine aminotransferase and aspartate aminotransferase levels. NASH was induced following 2 weeks in severe-phenotypic B6 albino (sB6), but B6 black exhibited no symptoms, even after 10 weeks. HCD-fed sB6 albino showed significantly higher mortality rate. Histological analysis of the liver revealed significant inflammatory cell and lipid infiltration and severe fibrosis. Serum lipoprotein analysis revealed significantly higher chylomicron and very low-density lipoprotein levels in sB6 albino. Moreover, significantly higher small intestinal lipid absorption and lower fecal lipid excretion occurred together with elevated intestinal NPC1L1 expression. As the tyrosinase point mutation represents the only genetic difference between B6 albino and B6 black, our work will facilitate the identification of susceptible genetic factors for NASH development and expand the understanding of NASH pathophysiology.

Scientific Reports     volume 11, Article number: 21827 (2021)
DOI 10.1038/s42003-021-02334-4

Research Outline (PDF In Japanese language)

Publication

Published:
2 Nov 2021

The transcriptional corepressor CtBP2 serves as a metabolite sensor orchestrating hepatic glucose and lipid homeostasis


Biological systems to sense and respond to metabolic perturbations are critical for the maintenance of cellular homeostasis. Here we describe a hepatic system in this context orchestrated by the transcriptional corepressor C-terminal binding protein 2 (CtBP2) that harbors metabolite-sensing capabilities. The repressor activity of CtBP2 is reciprocally regulated by NADH and acyl-CoAs. CtBP2 represses Forkhead box O1 (FoxO1)-mediated hepatic gluconeogenesis directly as well as Sterol Regulatory Element-Binding Protein 1 (SREBP1)-mediated lipogenesis indirectly. The activity of CtBP2 is markedly defective in obese liver reflecting the metabolic perturbations. Thus, liver-specific CtBP2 deletion promotes hepatic gluconeogenesis and accelerates the progression of steatohepatitis. Conversely, activation of CtBP2 ameliorates diabetes and hepatic steatosis in obesity. The structure-function relationships revealed in this study identify a critical structural domain called Rossmann fold, a metabolite-sensing pocket, that is susceptible to metabolic liabilities and potentially targetable for developing therapeutic approaches.

Nature Communications     12, Article number: 6315 (2021)
DOI 10.1038/s41467-021-26638-5

Research Outline (PDF In Japanese language)

New Publication

Published:
24 Sep 2021

A common genetic variant of a mitochondrial RNA processing enzyme predisposes to insulin resistance

CRISPR-Cas9 genome editing was used to introduce the missense N437S variant into the mouse Mrpp3 gene to study the causes of insulin resistance on a high-fat diet. The variant did not influence mitochondrial gene expression markedly, but instead, it reduced mitochondrial calcium that lowered insulin release from the pancreatic islet β cells of the Mrpp3 variant mice. Reduced insulin secretion resulted in lower insulin levels that contributed to imbalanced metabolism and liver steatosis.

The findings revealed that the MRPP3 variant may be a predisposing factor to insulin resistance and metabolic disease in the human population.


Science Advances - 24 Sep 2021 • Vol 7, Issue 39 •
    doi/10.1126/sciadv.abi7514

Publication

Published:
29 June 2021

Nuclear factor E2-related factor 2 (NRF2) deficiency accelerates fast fibre type transition in soleus muscle during space flight


Microgravity induces skeletal muscle atrophy, particularly in the soleus muscle, which is predominantly composed of slow-twitch myofibre (type I) and is sensitive to disuse. Muscle atrophy is commonly known to be associated with increased production of reactive oxygen species. However, the role of NRF2, a master regulator of antioxidative response, in skeletal muscle plasticity during microgravity-induced atrophy, is not known. To investigate the role of NRF2 in skeletal muscle within a microgravity environment, wild-type and Nrf2-knockout (KO) mice were housed in the International Space Station for 31 days. Gene expression and histological analyses demonstrated that, under microgravity conditions, the transition of type I (oxidative) muscle fibres to type IIa (glycolytic) was accelerated in Nrf2-KO mice without affecting skeletal muscle mass. Therefore, our results suggest that NRF2 affects myofibre type transition during space flight.

Communications Biology     volume 4, Article number: 787 (2021)
DOI 10.1038/s42003-021-02334-4
Nuclear factor E2-related factor 2 (NRF2) deficiency accelerates fast fibre type transition in soleus muscle during space flight.

Research Outline (PDF In Japanese language)

Publication

Published:
25 May 2021

Overexpression of Gata4, Mef2c, and Tbx5 Generates Induced Cardiomyocytes Via Direct Reprogramming and Rare Fusion in the Heart


(Note: Text has been auto translated from Japanese language.)
Cardiomyocytes that make up the heart have poor regenerative capacity and heart transplantation is the only radical treatment when cardiac function is significantly reduced. However, due to issues such as a shortage of donors, it is challenging to provide sufficient treatment. Regenerative medicine using pluripotent stem cells such as ES cells and iPS cells is attracting attention as an alternative method to heart transplantation, but it also leads to possibility of tumor formation, function issues and low tissue engraftment rate. The complexity and high cost of this approach is also a challenge to negate.


To date, this research group has developed a "myocardial direct reprogramming method" that induces cardiomyocytes directly from cardiac fibroblasts without using stem cells. This is another method that can help solve these issues. However, the myocardial cells produced by introducing a myocardial reprogramming gene into cardiac fibroblasts in vivo are truly induced regenerated myocardial cells derived from cardiac fibroblasts, or cardiac fibroblasts and surrounding myocardial cells.

It was unclear whether it was a myocardial cell that was being regenerated and formed by fusing. Therefore, in this study, we have shown that the new cardiomyocytes produced in vivo using genetically modified mice, are the genealogy and fusion of cells that are true regenerated cardiomyocytes derived from cardiac fibroblasts. This is the first time this has been revealed.

In this study, it was found that cardiomyocytes can be directly induced from cardiac fibroblasts by introducing a myocardial reprogramming gene into a mouse body. The results of this study greatly advance the realization of new cardiac regenerative medicine for heart disease.

Circulation     143:2123–2125(2021)
Overexpression of Gata4, Mef2c, and Tbx5 Generates Induced Cardiomyocytes Via Direct Reprogramming and Rare Fusion in the Heart


Research Outline (PDF In Japanese language)

Publication

Published:
11 June 2021

Development of technology to amplify hematopoietic stem cells in vitro and enable free gene transfer


(Note: Text has been auto translated from Japanese language.)

Hematopoietic stem cells have the ability to differentiate into various blood cells such as red blood cells, white blood cells, and platelets, and are being applied to the treatment of blood cancer and hereditary blood diseases. However, hematopoietic stem cells are rarely present in the bone marrow, and post-collection amplification is essential.


As a technique for mass amplification of hematopoietic stem cells derived from donor mice in vitro, a method using a medium supplemented with polyvinyl alcohol (PVA) has been reported. Therefore, this research team applied this method to maintain the function of hematopoietic stem cells from bone marrow cells containing a very small amount of hematopoietic stem cells by only immunomagnetic cell separation method using magnetic beads and subsequent simple culture operation. We have developed a technology that can be selectively purified and concentrated as it is. This facilitates gene transfer into hematopoietic stem cells outside the body, and at the same time, it can be amplified in large quantities, making it possible to transplant without using conventional pretreatment such as irradiation. Furthermore, by devising when introducing the target gene, by selecting and transplanting only the hematopoietic stem cells into which the gene has been correctly introduced, the target gene can be expressed in the blood cells in the mouse body at any time after transplantation. Was successful. With this method, there is no tissue damage caused by radiation to the recipient mice after transplantation, and stable breeding and observation can be performed for a long period of time. In addition, there is little invasion and tissue damage to living mice, and it is considered to be an excellent method from the viewpoint of animal welfare. By using these platforms, it has become possible to freely design the function of hematopoietic stem cells according to the purpose of treatment.

Nature Communications
12, Article number: 3568 (2021)     DOI 10.1038/s41467-021-23763-z
Non-conditioned bone marrow chimeric mouse generation using culture based enrichment of hematopoietic stem and progenitor cells.

Research Outline (PDF In Japanese language)