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Direct conversion of human fibroblasts into retinal pigment epithelium-like cells by defined factors

Protein & Cell 2014年第1期5卷 48-58页

作者： Kejing Zhang Guang-Hui Liu Fei Yi Nuria Montserrat Tomoaki Hishida concepcion rodriguez esteban Juan Carlos Izpisua Belmonte] Gene Expression Laboratory Salk Institute for Biological Studies 10010 North Torrey Pines Road La Jolla CA 92037 USA National Laboratory of Biomacromolecules Institute of Biophysics Chinese Academy of Sciences Beijing 100101 China Department of Molecular and Cellular Physiology Stanford University School of Medicine 265 Campus Drive StanfordCA 94305 USA Center for Regenerative Medicine in Barcelona Dr. Aiguader 88 08003 Barcelona Spain Biomedical Research Networking Center in Bioengineering Biomaterials and Nanomedicine (CIBER-BBN) BarcelonaSpain

The generation of functional retinal pigment epithelium （RPE） is of great therapeutic interest to the field of regenerative medicine and may provide possible cures for retinal degenerative diseases, including age-related macular degeneration （AMD）. Although RPE cells can be produced from either embryonic stem cells or induced pluripotent stem cells, direct cell reprogramming driven by lineage-determining transcription factors provides an immediate route to their generation. By monitoring a human RPE specific Bestl：：GFP reporter, we report the conversion of human fibroblasts into RPE lineage using defined sets of transcription factors. We found that Bestl：：GFP positive cells formed colonies and exhibited morphological and molecular features of early stage RPE cells. Moreover, they were able to obtain pigmen- tation upon activation of Retinoic acid （RA） and Sonic Hedgehog （SHH） signaling pathways. Our study not only established an ideal platform to investigate the tran- scriptional network regulating the RPE cell fate deter- mination, but also provided an alternative strategy to generate functional RPE cells that complement the useof pluripotent stem cells for disease modeling, drug screening, and cell therapy of retinal degeneration.

关键词： retinal pigment epithelium fibroblasts,direct conversion

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FOXO3-engineered human mesenchymal progenitor cells efficiently promote cardiac repair after myocardial infarction

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Protein & Cell 2021年第2期12卷 145-151页

作者： Jinghui Lei Si Wang Wang Kang Qun Chu Zunpeng Liu Liang Sun Yun Ji concepcion rodriguez esteban Yan Yao Juan Carlos Izpisua Belmonte Piu Chan Guang-Hui Liu Weiqi Zhang Moshi Song Jing Qu Advanced Innovation Center for Human Brain Protection National Clinical Research Center for Geriatric DisordersXuanwu Hospital Capital Medical UniversityBeijing 100053China State Key Laboratory of Membrane Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China Disease Genomics and Individualized Medicine Laboratory Beijing Institute of GenomicsChinese Academy of SciencesBeijing 100101China Institute of Stem Cell and Regeneration Chinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China The MOH Key Laboratory of Geriatrics Beijing HospitalNational Center of GerontologyBeijing 100730China Department of Cardiology Beijing Anzhen HospitalCapital Medical UniversityBeijing 100029China Gene Expression Laboratory Salk Institute for Biological StudiesLa Jolla 92037USA

Dear Editor,Myocardial infarction(MI)is the irreversible cardiomyocyte death resulting from prolonged oxygen deprivation due to obstructed blood supply(ischemia),leading to contractile dysfunction and cardiac *** recent decades,stem cell transplantation has been extensively investigated for the repair of injured heart in animal studies and clinical trials(Kanelidis et al.,2017;Gyongyosi et al.,2018).

关键词： al. infarction cardiac

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Global DNA methylation and transcriptional analyses of human ESC-derived cardiomyocytes

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Protein & Cell 2014年第1期5卷 59-68页

作者： Ying Gut Guang-Hui Liu Nongluk Plongthongkum' Christopher Benner Fei Yi Jing Qu Keiichiro Suzuki Jiping Yang Weiqi Zhang Mo Li Nuria Montserrat Isaac Crespo Antonio del Sol concepcion rodriguez esteban Kun Zhang Juan Carlos Izpisua Belmonte Gene Expression Laboratory Salk Institute for Biological Studies 10010 North Torrey Pines Road La Jolla CA 92037 USA National Laboratory of Biomacromolecules institute of Biophysics Chinese Academy of Sciences Beijing 100101 China ' State key Laboratory of Drug Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai 201203 China Department of Bioengineering University of California at San Diego La Jolla CA 92093 USA center of Regenerative Medicine in Barcelona Dr. Aiguader 88 08003 Barcelona Spain Biomedical Research Networking Center in Bioengineering Biomaterials and Nanomedicine (CIBER-BBN) Poeta MarianoEsquillor s/n 50018 Sargossa Spain Luxembourg Centre for Systems Biomedicine (LCSB) University of Luxembourg Walferdange Luxembourg

With defined culture protocol, human embryonic stem cells （hESCs） are able to generate cardiomyocytes in vitro, therefore providing a great model for human heart development, and holding great potential for car- diac disease therapies. In this study, we successfully generated a highly pure population of human cardio- myocytes （hCMs）（〉95% cTnT＋） from hESC line, which enabled us to identify and characterize an hCM-specific signature, at both the gene expression and DNA meth- ylation levels. Gene functional association network and gene-disease network analyses of these hCM-enriched genes provide new insights into the mechanisms of hCM transcriptional regulation, and stand as an informative and rich resource for investigating cardiac gene func- tions and disease mechanisms. Moreover, we show that cardiac-structural genes and cardiac-transcription fac- tors have distinct epigenetic mechanisms to regulate their gene expression, providing a better understandingof how the epigenetic machinery coordinates to regulate gene expression in different cell types.

关键词： human cardiomyocyte DNA methylation,microarray heart development

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A widely adaptable approach to generate integration-free iPSCs from non-invasively acquired human somatic cells

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Protein & Cell 2015年第5期6卷 386-389页

作者： Zhichao Ding Lina Sui Ruotong Ren Yanjun Liu Xiuling Xu Lina Fu Ruijun Bai Tingting Yuan Ying Hao Weiqi Zhang Huize Pan Wensu Liu Han Yu concepcion rodriguez esteban Xiaobing Yu Ze Yang Jian Li Xiaomin Wang Juan Carlos Izpisua Belmonte Guang-Hui Liu Fei Yi Jing Qu National Laboratory of Biomacromolecules Institute of Biophysics University of the Chinese Academy of Sciences Chinese Acad- emy of Sciences Beijing 100101 China Department of Endocrinology 306th Hospital of PLA Beijing100101 China Science and Development Xtem Foshan 528000 China State Key Laboratory of Reproductive Biology Institute of ZoologyChinese Academy of Sciences Beijing 100101 China Beijing Hospital of the Ministry of Health Beijing 100073 China Beijing Institute for Brain Disorders Beijing 100069 China Gene Expression Laboratory Salk Institute for Biological Studies10010 North Torrey Pines Road La Jolla CA 92037 USA Center for Molecular and Translational Medicine Beijing 100101China Department of Molecular and Cellular Physiology StanfordUniversity School of Medicine Stanford CA 94305 USA

Dear Editor Human pluripotent stem cells including human embryonic stem cells （ESCs） and induced pluripotent stem cells （iPSCs） are cells displaying abilities of unlimited self-renewal and differentiation into any somatic cell type. These unique properties make them increasingly attractive for novel applications in disease modeling, drug discovery, and cell therapy （Buganim et al., 2014; Liu et al., 2011; Liu et al., 2012; Sanchez Alvarado and Yamanaka, 2014）. Moreover, iPSCs hold great potential for personalized cell therapy as they avoid some of the ethical concerns as well as the immunological rejection issues ascribed to ESCs.

关键词： adaptable approach generate integration-free iPSCs

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Mutations in foregut SOX2^+ cells induce efficient proliferation via CXCR2 pathway

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Protein & Cell 2019年第7期10卷 485-495页

作者： Tomoaki Hishida Eric Vazquez-Ferrer Yuriko Hishida-Nozaki Ignacio Sancho-Martinez Yuta Takahashi Fumiyuki Hatanaka Jun Wu Alejandro Ocampo Pradeep Reddy Min-Zu Wu Laurie Gerken Reuben J. Shaw concepcion rodriguez esteban Christopher Benner Hiroshi Nakagawa Pedro Guillen Garcia Estrella Nunez Delicado Antoni Castells Josep M. Campistol Guang-Hui Liu Juan Carlos Izpisua Belmonte Gene Expression Laboratory Salk Institute for Biological Studies 10010 North Torrey Pines Road La Jolla CA 92037 USA Advaneed Innovation Center for Human Brain Protection National Clinical Research Center for Geriatric Disorders Xuanwu Hospital Capital Medical University Beijing 100053 China National Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences Beijing 100101 China University of the Chinese Academy of Sciences Beijing 100049 China Insitute for Stem Cell and Regeneration Chinese Academy of Sciences Beijing 100101 China Beijing Institute for Brain Disorder Beijing 100069 China Universidad Catolica San Antonio de Murcia Campus de los Jeronimos 135 Guadalupe 30107 Spain Molecular and Cell Biology Laboratory Dulbecco Center for Cancer Research Salk Institute for Biological Studies 10010 North Torrey Pines Road La Jolla CA 92037 USA Howard Hughes Medical Institute Dulbecco Center for Cancer Research Salk Institute for Biological Studies 10010 North Torrey Pines Road La Jolla CA 92037 USA Integrative Genomics Core Salk Institute for Biological Studies 10010 North Torrey Pines Road La Jolla CA 92037 USA Division of Gastroenterology Department of Medicine Perelman School of Medicine University of Pennsylvania Philadelphia PA 19104 USA Abramson Cancer Center University of Pennsylvania Philadelphia PA 19104 USA Department of Traumatology and Research Unit Clinica CEMTRO Av. Ventisquero de la Condesa 42 Madrid 28035 Spain Gastroenterology Department Hospital Clinic University of Barcelona IDIBAPS CIBEREHD Barcelona 08036 Spain

Identification of the precise molecular pathways involved in oncogene-induced transformation may help us gain a better understanding of tumor initiation and promotion. Here, we demonstrate that SOX2^+ foregut epithelial cells are prone to oncogenic transformation upon mutagenic insults, such as Kras^G12D and p53 deletion. GFP-based lineage-tracing experiments indicate that SOX2^+ cells are the cells-of-origin of esophagus and stomach hyperplasia. Our observations indicate distinct roles for oncogenic KRAS mutation and P53 deletion. p53 homozygous deletion is required for the acquisition of an invasive potential, and Kras^G12D expression, but not p53 deletion, suffices for tumor formation. Global gene expression analysis reveals secreting factors upregulated in the hyperplasia induced by oncogenic KRAS and highlights a crucial role for the CXCR2 pathway in driving hyperplasia. Collectively, the array of genetic models presented here demonstrate that stratified epithelial cells are susceptible to oncogenic insults, which may lead to a better understanding of tumor initiation and aid in the design of new cancer therapeutics.

关键词： Sox2 tumor CXCR2 stratified epithelia

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αKLOTHO and sTGFβR2 treatment counteract the osteoarthritic phenotype developed in a rat model

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Protein & Cell 2020年第3期11卷 219-226页

作者： Paloma Martinez-Redondo Isabel Guillen-Guillen Noah Davidsohn Chao Wang Javier Prieto Masakazu Kurita Fumiyuki Hatanaka Cuiqing Zhong Reyna Hernandez-Benitez Tomoaki Hishida Takashi Lezaki Akihisa Sakamoto Amy NNemeth Yuriko Hishida concepcion rodriguez esteban Kensaku Shojima Ling Huang Maxim Shokhirev Estrella Nunez-Delicado Josep MCampistol Isabel Guillen-Vicente Elena rodriguez-Inigo Juan Manuel Lopez-Alcorocho Marta Guillen-Vicente George Church Pradeep Reddy Pedro Guillen-Garcia Guang-Hui Liu Juan Carlos Izpisua Belmonte Gene Expression Laboratory Salk Institute for Biological Studies10010 North Torrey Pines RoadLa JollaCA 92037USA Wyss Institute for Biologically Inspired Engineering Harvard UniversityCambridgeMA 02115USA Department of Genetics Harvard Medical SchoolBostonMA 02115USA Integrative Genomics and Bioinformatics Core Salk Institute for Biological Studies10010 North Torrey Pines RoadLa JollaCA 92037USA Universidad Catolica San Antonio de Murcia(UCAM) Campus de los JeronimosNo 1351230107 GuadalupeSpain Hospital Clinic of Barcelona Career Villarroel17008036 BarcelonaSpain Department of Traumatology and Research Unit Clinica CEMTRO28035 MadridSpain National Laboratory of Biomacromolecules CAS Center for Excellence in BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Institute for Stem cell and Regeneration Chinese Academy of SciencesBeijing 100101China Translational Medical Center for Stem Cell Therapy Shanghai East HospitalTongji University School of MedicineShanghai 200120China

Dear Editor,Homeostasis and repair are critical biological processes that allow for tissue and organ preservation and function in multicellular *** regulation and extension vary drastically across the animal kingdom,and mammals show limited tissue-specific regenerative capacity that declines with *** aging,articular cartilage is one of the tissues that undergo substantial changes in the matrix structure,molecular composition,metabolic activity,and mechanical properties(Loeser et al.2016).

关键词： aging treatment critical

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Open avenues for carotenoid biofortification of plant tissues

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Plant Communications 2023年第1期4卷 228-237页

作者： Luca Morelli Manuel rodriguez-concepcion Institute for Plant Molecular and Cell Biology(IBMCP) CSIC-Universitat Polite`cnica de Vale`ncia46022 ValenciaSpain Department of Biology&CESAM&ECOMARE Universidade de Aveiro3810-193 AveiroPortugal

Plant carotenoids are plastidial isoprenoids that function as photoprotectants,pigments,and precursors of apocarotenoids such as the hormones abscisic acid and *** do not produce carotenoids but need to obtain them from their diet as precursors of retinoids,including vitamin *** also provide numerous other health *** attempts to improve the carotenoid profile of different crops have been carried out by manipulating carotenoid biosynthesis,degradation,and/or ***,we will focus on open questions and emerging subjects related to the use of biotechnology for carotenoid *** impressive achievements,new efforts should be directed to extend the use of genome-editing technologies to overcome regulatory constraints and improve consumer acceptance of the carotenoid-enriched *** challenge is to prevent off-target effects like those resulting from altered hormone levels and metabolic *** on biofortification of green tissues should also look for new ways to deal with the negative impact that altered carotenoid contents may have on *** a carotenoid-enriched product has been obtained,additional effort should be devoted to confirming that carotenoid intake from the engineered food is also *** involves ensuring post-harvest stability and assessing bioaccessibility of the biofortified product to confirm that release of carotenoids from the food matrix has not been negatively *** addressing these challenges will ensure new milestones in carotenoid biotechnology and biofortification.

关键词： biofortification biotechnology carotenoids plastidial isoprenoids vitamin A

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Plant geranylgeranyl diphosphate synthases: every (gene) family has a story

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aBIOTECH 2021年第3期2卷 289-298页

作者： M.Victoria Barja Manuel rodriguez-concepcion Centre for Research in Agricultural Genomics(CRAG)CSIC-IRTA-UAB-UB Campus UAB Bellaterra08193 Barce-lonaSpain Institute for Plant Molecular and Cell Biology(BMCP) CSIC-Universitat Politecnica de Valencia46022 ValenciaSpain

Plant isoprenoids(also known as terpenes or terpenoids)are a wide family of primary and secondary metabolites with multiple *** particular,most photosynthesis-related isoprenoids(including carotenoids and chlorophylls)as well as diterpenes and polyterpenes derive from geranylgeranyl diphosphate(GGPP)produced by GGPP synthase(GGPPS)enzymes in several cell *** genomes typically harbor multiple copies of differentially expressed genes encoding GGPPS-like *** sequence comparisons allow to identify potential GGPPS candidates,experimental evidence is required to ascertain their enzymatic activity and biologi cal ***,functional analyses of the full set of potential GGPPS paralogs are only available for a handful of plant *** we review our current knowledge on the GGPPS families of the model plant Arabidopsis thaliana and the crop species rice(0ryza sativa),pepper(Capsicum annuum)and tomato(Solanum lycopersicum).The results indicate that a major determinant of the biological role of particular GGPPS paralogs is the expression profile of the corresponding genes even though specific interactions with other proteins(including GGPP-consuming enzymes)might also contribute to *** some species,however,a single GGPPS isoforms appears to be responsible for the production of most if not all GGPP required for cell *** the mechanisms regulating GGPPS activity in particular cell compartments,tissues,organs and plant species will be very useful for future metabolic engineering approaches aimed to manipulate the accumulation of particular GGPP-derived products of interest without negatively impacting the levels of other isoprenoids required to sustain essential cell functions.

关键词： Geranylgeranyl diphosphate GGPP GGPP synthase Isoprenoid Isoprenyl transferase

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New Insights into Plant Isoprenoid Metabolism

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Molecular Plant 2012年第5期5卷 964-967页

作者： Pablo Pulido Catalina Perello Manuel rodriguez-concepcion Department of Molecular Genetics Centre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB Campus UAB Bellaterra 08193 BarcelonaSpain

Isoprenoids are a hugely diverse family of compounds derived from the C5 precursors isopentenyl diphosphate （IPP） and dimethylallyl diphosphate （DMAPP）. Although all free-living organisms synthesize isoprenoids, they are particularly abun- dant and diverse in plants, with tens of thousands structures known to date. The highest variety of plant isoprenoids are specialized （secondary） metabolites that participate in the interaction of plants with their environment. These include pigments, volatiles, and defense compounds, some of which have applications in industry and agriculture.

关键词： Metabolism Insight Plant

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SNOWY COTYLEDON 2 Promotes Chloroplast Development and Has a Role in Leaf Variegation in Both Lotus japonicus and Arabidopsis thaliana

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Molecular Plant 2017年第5期10卷 721-734页

作者： Nicola Zagari Omar Sandoval-lbanez Niels Sandal Junyi Su Manuel rodriguez-concepcion Jens Stougaard Mathias Pribil Dario Leister Pablo Pulido Plant Molecular Biology Department of Biology I Ludwig-Maximilians-Universitat Munchen 82152 Planegg-Martinsried Germany Copenhagen Plant Science Center University of Copenhagen 1871 Frederiksberg C Denmark Research and Innovation Center Fondazione Edmund Mach via E. Mach 1 38010 San Michele all'Adige Italy Centre for Carbohydrate Recognition and Signalling Department of Molecular Biology and Genetics Aarhus University Gustav Wieds Vej 10 8000 Aarhus C Denmark Centre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB Campus UAB Bellaterra 08193 Barcelona Spain

Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essential for chloroplast development and the transition from heterotrophic to phototrophic growth. Snowy cotyledon 2 （SCO2） is a DNAJ-like protein involved in thylakoid membrane biogenesis and interacts with the light-harvesting chlorophyll-binding protein LHCBI. In Arabidopsis thaliana, SCO2 function was previ- ously reported to be restricted to cotyledons. Here we show that disruption of SC02 in Lotus japonicus results not only in paler cotyledons but also in variegated true leaves. Furthermore, smaller and pale- green true leaves can also be observed in A. thaliana sco2 （atsco2） mutants under short-day conditions. In both species, SCO2 is required for proper accumulation of PSlI-LHCll complexes. In contrast to other variegated mutants, inhibition of chloroplastic translation strongly affects L. japonicus sco2 mutant devel- opment and fails to suppress their variegated phenotype. Moreover, inactivation of the suppressor of variegation AtClpR1 in the atsco2 background results in an additive double-mutant phenotype with variegated true leaves. Taken together, our results indicate that SCO2 plays a distinct role in PSll assembly or repair and constitutes a novel factor involved in leaf variegation.

关键词： SCO2 DNAJ-like photosynthesis assembly factor variegation chloroplast development

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