Publications

♦ 2018 Publications:

♠ Shaw-Fang Yet 林秀芳博士

Redox Biol. 2018 May;15:51-61. doi: 10.1016/j.redox.2017.11.019. Epub 2017 Nov 22.

PMID: 29216542

Loss Of Heme Oxygenase-1 Accelerates Mesodermal Gene Expressions During Embryoid Body Development From Mouse Embryonic Stem Cells.

Heme oxygenase (HO)-1 is an inducible stress response protein and well known to protect cells and tissues against injury. Despite its important function in cytoprotection against physiological stress, the role of HO-1 in embryonic stem cell (ESC) differentiation remains largely unknown. We showed previously that induced pluripotent stem (iPS) cells that lack HO-1 are more sensitive to oxidant stress-induced cell death and more prone to lose pluripotent markers upon LIF withdrawal. To elucidate the role of HO-1 in ESC differentiation and to rule out the controversy of potential gene flaws in iPS cells, we derived and established mouse HO-1 knockout ESC lines from HO-1 knockout blastocysts. Using wild type D3 and HO-1 knockout ESCs in the 3-dimensional embryoid body (EB) differentiation model, we showed that at an early time point during EB development, an absence of HO-1 led to enhanced ROS level, concomitant with increased expressions of master mesodermal regulator brachyury and endodermal marker GATA6. In addition, critical smooth muscle cell (SMC) transcription factor serum response factor and its coactivator myocardin were enhanced. Furthermore, HO-1 deficiency increased Smad2 in ESCs and EBs, revealing a role of HO-1 in controlling Smad2 level. Smad2 not only mediates mesendoderm differentiation of mouse ESCs but also SMC development. Collectively, loss of HO-1 resulted in higher level of mesodermal and SMC regulators, leading to accelerated and enhanced SMC marker SM α-actin expression. Our results reveal a previously unrecognized function of HO-1 in regulating SMC gene expressions during ESC-EB development. More importantly, our findings may provide a novel strategy in enhancing ESC differentiation toward SMC lineage. https://www.sciencedirect.com/science/article/pii/S2213231717306158?via%3Dihub

♠ Betty Lin-Ju Yen, M.D 顏伶汝博士

NHRI researchers report differentiation of MSCs from iPSCs in downregulation of c‐Myc and DNA replication pathways with immunomodulation toward CD4 and CD8 cells

人類誘導性多功能幹細胞衍生之間質幹細胞具有低致癌性及調控異常免疫反應的應用潛力

Stem Cells 36:903-914, 2018 (http://dx.doi.org/10.1002/stem.2795)

Multilineage tissue‐source mesenchymal stem cells (MSCs) possess strong immunomodulatory properties and are excellent therapeutic agents, but require constant isolation from donors to combat replicative senescence. The differentiation of human induced pluripotent stem cells (iPSCs) into MSCs offers a renewable source of MSCs; however, reports on their immunomodulatory capacity have been discrepant. A team led by Dr. B. Linju Yen from the Institute of Cellular and System Medicine, in collaboration with Dr. Ko-Jiunn Liu, National Institute of Cancer Research and Vice President Dr. Huey-Kang Sytwu, from National Institute of Infectious Diseases and Vaccinology, recently published their findings in Stem Cells (2018 Feb 3. doi: 10.1002/stem.2795), supporting that iPSC‐MSCs possess low oncogenicity and strong immunomodulatory properties regardless of cell‐of‐origin or reprogramming method and are good potential candidates for therapeutic use. Using MSCs differentiated from iPSCs reprogrammed using diverse cell types and protocols, and in comparison to human embryonic stem cell (ESC)‐MSCs and bone marrow (BM)‐MSCs, the team performed transcriptome analyses and assessed for functional immunomodulatory properties.

人類的間質幹細胞(mesenchymal stem cells; MSCs)是一種具有多分化潛力及免疫調控能力的成體幹細胞,故擁有高度臨床應用性。但其數目在體內較為稀少,而幹細胞治療則需要較多的細胞數目(每次約需108~1010細胞),因此MSCs取得後皆需在體外培養,將細胞數量放大才能達到臨床使用需求。但這個體外培養的過程往往會產生複製性衰老(replicative senescence),因而降低幹細胞的增殖、分化、及免疫調控功能,這一點是MSCs在臨床應用上的一大瓶頸。因此如果利用與人類胚胎幹細胞(human embryonic stem cells; hESCs)相似具高度增生能力,但又可“客製化”且的人類誘導性多功能幹細胞(induced pluripotent stem cells; iPSCs)來做為細胞材料來源,將其源源不絕地衍生出下游的MSCs,則不但可以解決成體幹細胞體外培養會衰老的問題,同時因已分化至MSC,故也可避開直接使用iPSCs及hESCs可能在體內會產生畸胎瘤(teratoma)的疑慮。

本所顏伶汝醫師的團隊,之前已成功建立多株台灣本土的人類iPSCs,此次則是經由顏醫師、本院癌症研究所劉柯俊博士以及感染及免疫研究所司徒惠康副院長共同合作,發現人類iPSC-MSCs,不論是使用何種再程序化方式(reprogramming method),或何種細胞來源(cell source)得到的iPSCs,其所衍生出來的MSC都具有多分化潛力。而且它們的免疫調控能力與骨髓間葉幹細胞及由hESC衍生而來的MSCs(hESC-MSCs),相較起來並無差異。此研究成果已發表於 2018年2月的Stem Cells期刊 [2018 Feb 3. doi: 10.1002/stem.2795]。

 

♠ Hua-Jung Li, 李華容博士

Lin MC, Chen SY, He PL, Herschman HR, and Li HJ* PGE2/EP4 antagonism enhances tumor chemosensitivity by inducing extracellular vesicle‐mediated clearance of cancer stem cells. Int J Cancer. 2018;15;143(6):1440-1455 (IF 2017: 7.36, 24/217, ONCOLOGY).

♦ 2017 Publications:

♠ Jeng-Jiann Chiu 裘正健博士

NHRI researchers report microRNA-10a is crucial for endothelial response to different flow patterns via interaction of retinoid acid receptors and histone deacetylases

微小核糖核酸-10a藉由維甲酸受體及組織蛋白去乙醯酶的互動調控血管內皮接受不同血液動力學作用時的病生理反應

http://enews.nhri.org.tw/enews_list_new2_more.php?volume_indx=714&showx=showarticle&article_indx=11891

♠ Hua-Jung Li, 李華容博士

Lin MC, Chen SY, Tsai HM, He PL, Lin YC, Herschman HR, and Li HJ* PGE2/EP4 signaling controls the transfer of the mammary stem cell state by lipid rafts in extracellular vesicles. Stem Cells 2017;35(2):425-444 (IF 2016: 6.281, 14/161, BIOTECHNOLOGY & APPLIED MICROBIOLOGY).

Lin MC, Chen SY, He PL, Luo WT, and Li HJ* Transfer of Mammary Gland-forming Ability Between Mammary Basal Epithelial Cells and Mammary Luminal Cells via Extracellular Vesicles/Exosomes. J. Vis. Exp. 2017; 124, e55736, doi:10.3791/55736 (IF 2016: 1.659, 28/64, MULTIDISCIPLINARY SCIENCES)

2016 Publications:

♠ Betty Lin-Ju Yen, M.D 顏伶汝博士

Human pluripotent stem cell (PSC)-derived mesenchymal stem cells (MSCs) show potent neurogenic capacity which is enhanced with cytoskeletal rearrangement 人類多功能幹細胞之間質幹細胞透過骨架重組增強神經分化的能力

Oncotarget 7:43949-43959, 2016 (doi: 10.18632/oncotarget.9947)

Stem cells therapy provides a therapeutic strategy for neuro-degenerative diseases due to generate functional neuron. Post-natal mesenchymal stem cells (post-natal MSCs) have capacities of neuro-lineage differentiation and immunomodulatory, which offer the possibility for therapeutic use in neuro-degenerative disease. However, in order to support high cell volume for cell therapy, post-natal MSCs expanses in vitro which easily result in replicative senescence. MSCs derived from pluripotent stem cells (PSC), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPS), can overcome this issue by enhanced proliferative ability. We found that the development stage of human MSCs affected the outcome of lineage commitment as mediated by cytoskeletal rearrangement. The ROCK inhibitor, Y-27632, in human pluripotent stem cells-MSCs (PSC-MSCs) efficiently pushed these developmentally early-stage MSCs towards a neuron progenitor (NP) phenotype. PSC-MSCs treated with Y-27632 do not appear mesodermal differentiation, but rather showed a neuron-like phenotype, with rapid development of cell processes while preserving cell viability. Moreover, Y-27632-treated PSC-MSCs expressed several neuron-restricted progenitor (NRP) markers included β-III-tubulin, Doublecortin, and NeuN at protein level. Our data not only demonstrate the important role of the cytoskeleton in MSC lineage commitment, but also the influence of the developmental stages of stem cells on the subsequent differentiation.

圖一: PSC-MSCs 在神經分化培養液中抑制ROCK作用,更能促進神經分化 Nestin (綠色螢光),Doublecortin (紅色螢光),Y (Y-27632, ROCK inhibitor),NDM (neurogenic differentiation medium), SF+Y(serum free+Y-27632). Scale bar: 20μm.

Figure 1: Inhibition of ROCK in PSC-MSCs lead to more committed differentiation into neural-like cells. Y (Y-27632, ROCK inhibitor),NDM (neurogenic differentiation medium), SF+Y(serum

間質幹細胞(mesenchymal stem cells; MSC)為俱有免疫調節特性之組織幹細胞。間葉幹細胞為硬骨、軟骨、及脂肪組織的幹細胞,但也被發現在特定的調控下可分化為神經細胞。目前間葉幹細胞最常見的來源為從成體的骨髓(bone marrow MSCs; BMMSCs)及脂肪分離,但成體來源的幹細胞數目稀少,需要體外大量繼代培養才可有足夠的數量進行臨床應用,而在體外培養時會造成成體細胞的老化。人類多功能幹細胞(human pluripotent stem cells; PSCs)包含人類胚胎幹細胞(human embryonic stem cells; hESCs)及誘導性幹細胞(induced pluripotent stem cells; iPSCs),提供兩全其美的間質幹細胞來源。本院細胞及系統醫學研究所顏伶汝研究員及其研究團隊發現PSC-MSCs較成體間葉幹細胞俱有更強的神經分化能力。與骨髓間質幹細胞比較,PSC-MSCs高度表現神經幹細胞(neural stem cells) 相關的基因及轉錄因子,且透過神經分化培養液 (neurogenic differentiation medium, NDM)或抑制細胞骨架調控激酶蛋白(ROCK)能迅速分化成似神經細胞 (neural-like cell)。最重要是,當在NDM狀態下同時抑制ROCK造成細胞骨架重組,PSC-MSCs可分化成較後期神經細胞,此細胞俱有有絲分裂後期神經細胞 (post-mitotic neurons)特徵,包括細胞核縮小,樹突的延展及表現特定神經前驅細胞(neuron-restricted progenitor)基因,如NeuN, β-III-tubulin 和 Doublecortin (圖一)。此研究也是第一篇論文應用到由生技醫藥國家型科技計畫(NRPB)補助之人類疾病誘導型多潛能幹細胞服務聯盟資源中心(Human Disease iPSCs Service Consortium)所建立的iPSC細胞株。此資源中心的目標為建立及提供本土之人類iPSCs,應用於相關研究及建立iPSC測試疾病及藥物之平台,以及研究幹細胞治療之細胞。

本研究的結果顯示PSC-MSCs俱有分化成神經細胞的高度潛力,對於神經退化性疾病治療提供希望。此研究發表在2016年六月的Oncotarget期刊 (2016 Jun 11. doi: 10.18632/oncotarget.9947.)

圖二:利用調控細胞骨架,可造成PSC-MCSs表現特定神經前驅細胞(neuron-restricted progenitor)基因及相似的細胞形態《文/圖:細胞及系統醫學研究所顏伶汝研究員、彭凱彥博士班學生》

2015 Publications:

Interleukin-25 Mediates Transcriptional Control of PD-L1 via STAT3 in Multipotent Human Mesenchymal Stromal Cells (hMSCs) to Suppress Th17 Responses.

間葉幹細胞透過IL-25/PD-L1治療Th17相關自體免疫疾病的新曙光

Stem Cell Reports 5:392-404, 2015

Multipotent human mesenchymal stem cells (hMSCs) harbor immunomodulatory properties which are therapeutically relevant. One of the most clinically important populations of leukocytes is the interleukin-17A (IL-17A)-secreting T (Th17) lymphocytes, which are implicated in many human pathologies including autoimmune diseases and chronic inflammatory states. However, mechanisms of hMSC and Th17 cell interactions are incompletely resolved. Dr. B. Linju Yen’s lab demonstrated that hMSCs strongly suppress Th17 responses through expression of IL-25, a paracrine factor, and programmed death ligand-1 (PD-L1), a cell surface ligand critical to T cell tolerance induction. The requirement of both factors is explained by IL-25 modulation of PD-L1 expression via JNK and STAT3 to orchestrate an overall effect of suppressing Th17 responses (Figure 1). These findings demonstrate the complexities of hMSC-mediated Th17 suppression, as well as highlight the IL-25/STAT3/PD-L1 axis as a candidate therapeutic target. The article was published in Stem Cell Reports and was a highlighted article, with ongoing patent application and technology transfer of this discovery.

Fig 1. Schematic shows a model of hMSC-mediated suppression of Th17 responses involving the IL-25/STAT3/PD-L1 axis.

人類間葉幹細胞(簡稱MSCs)是目前被認為最有臨床應用性的幹細胞種類之一,此類組織幹細胞不但具有多種細胞分化能力,也有強烈的免疫抑制功能,尤其是針對T淋巴細胞。本所顏伶汝醫師實驗室發現利用間葉幹細胞分泌介白素-25(IL-25)因子,經由JNK訊息傳遞及STAT3轉錄因子,增加間葉幹細胞本生細胞膜表面PD-L1 的表現而抑制一種會分泌介白素17A(IL-17A)的特殊CD4 T淋巴細胞(又稱為Th17細胞)之反應(圖一)。Th17細胞在多種疾病中均有重要角色,包含許多種自體免疫疾病、慢性發炎疾病、以及癌症,而此研究為間葉幹細胞與Th17細胞間的互動找出清楚的路徑,是幹細胞研究的又一創新成果本研究之成果而也可以提供未來直接利用間葉幹細胞或藉由調控間葉幹細胞分泌IL-25之功能,來治療因過度Th17細胞反應而造成之相關免疫疾病。這些結果於2015發表於CELL系列之國際期刊Stem Cell Reports,而且是當月被highlight的論文,備受國際重視,而此發現也目前進行申請專利及技轉中。

圖一:經由JNK訊息傳遞及STAT3轉錄因子,增加間葉幹細胞本生細胞膜表面PD-L1的表現,而抑制一種會分泌介白素17A(IL-17A)的特殊CD4 T淋巴細胞(又稱為Th17細胞)之反應。

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