Research Interests

The research of the Functional Genomics and RNA-based Therapeutics laboratory is focused on two main areas: i) the identification and molecular characterization of novel cellular factors relevant to cardiac and neuronal regeneration, and the translation of this knowledge into effective RNA-based therapeutic strategies, and ii) the development of high-throughput and high-content screening technologies using genome-wide siRNA and microRNA libraries.

• Novel RNA-based therapeutics for cardiac and neuronal regeneration

  Cardiovascular diseases, including myocardial infarction and cerebral ischemic stroke, are the leading cause of death globally. The main area of research of the Functional Genomics and RNA-based Therapeutics laboratory is the identification and characterization of novel cellular factors that control regeneration of cardiac and neuronal tissues following injury.

  This will be achieved through the use of systematic and unbiased genome-wide experimental approaches such as functional high-throughput screening (HTS) and RNA-seq. The identified factors (genes and microRNAs) will be characterized extensively in terms of their mechanisms of action, and the effect of modulating these factors will be tested in relevant animal models.

  Our ultimate goal is to exploit this knowledge to develop novel RNA-based therapeutic strategies able to induce heart and brain regeneration.

  

• High-Throughput and High-Content Screening using genome-wide siRNA and microRNA libraries

  Another important area of research of the Functional Genomics and RNA-based Therapeutics laboratory is the development of high-throughput screening (HTS) and high-content screening (HCS) technologies and their application to different areas of biomedical research (e.g. regenerative medicine, infection biology, intracellular signaling and cancer).

  The state-of-the-art high-throughput screening (HTS) platform hosted by the Functional Genomics and RNA-based Therapeutics laboratory at CNC is the first platform in Portugal equipped with the adequate instrumentation, libraries and expertise to perform human and mouse genome-wide siRNA and microRNA screenings.

  

Miguel Mano, PhD Group Leader Investigator

Vagner Rigaud PhD Post-Doctoral Fellow

Mariana Bexiga PhD Post-Doctoral Fellow

Ricardo Silva MSc PhD Student PDBEB

Inês Lopes MSc Research Fellow

Inês Caramelo MSc Research Fellow

Highly motivated candidates (undergraduates, prospective PhD students and Postdocs) interested in joining the Functional Genomics and RNA-based Therapeutics laboratory are encouraged to contact the PI directly.

High-Throughput Screening (HTS) Plataform

The Functional Genomics and RNA-based Therapeutics laboratory at CNC hosts a state-of-the-art high-throughput screening (HTS) platform, in a BSL2 environment, equipped with the adequate instrumentation, libraries and expertise to perform genome-wide siRNA and microRNA screenings in human and mouse cellular models.

Libraries available for screening:
- human genome-wide siRNA
- mouse genome-wide siRNA
- human microRNA mimics
- human microRNA inhibitors

Collaborations

Mauro Giacca
International Centre for Genetic Engineering and Biotechnology (ICGEB, Trieste, Italy)

Ana Eulálio
Center for Neuroscience and Cell Biology (CNC, Coimbra, Portugal)
Institute for Infection Biology (IMIB, Wuerzburg, Germany)

Giannino Del Sal
National Laboratory CIB (Trieste, Italy)

Cynthia Sharma
Research Center for Infectious Diseases (ZINF, Wuerzburg, Germany)

Daniel Lopez
National Centre for Biotechnology (CNB/CSIC, Madrid, Spain)

Funding

Designação do projeto:  IR-miRNA : Identificação e caracterização funcional de microRNAs reguladores de dano cardíaco por isquemia-reperfusão

Código do projeto: POCI-01-0145-FEDER-016696
Região de intervenção: Centro - Coimbra Entidade Beneficiária:  CNC - Centro de Neurociências e Biologia Celular

Custo total elegível: 177 540,00 €

Apoio financeiro da União Europeia: FEDER – 150 909,00 €

Apoio financeiro público nacional/regional: FCT – 26 631,00 €

Objetivo Principal:
O dano por isquemia-reperfusão é uma das mais importantes causas de morbilidade e mortalidade, transversal a várias patologias e cenários clínicos, incluindo infarto do miocárdio. A isquemia é caracterizada por uma insuficiente irrigação dos tecidos, privando-os de oxigénio. Paradoxalmente, o dano causado por isquemia prolongada é ulteriormente agravado durante a fase de reperfusão, originando um dano irreversível. A identificação de factores que contribuem para o dano causado por isquemia-reperfusão é essencial para evitar ou diminuir a extensão deste, e potencialmente desenvolver novas abordagens terapêuticas para intervenção em situações de infarto do miocárdio.

MicroRNA (miRNAs) são RNAs endógenos não codificantes de pequenas dimensões, que desempenham um papel importante na regulação da expressão génica ao nível pós-transcripcional. Estudos recentes demonstraram que os miRNAs exercem um papel regulatório importante em diversos aspectos relacionados com doenças cardiovasculares, incluindo o dano por isquemia-reperfusão. Apesar de um pequeno número de miRNAs ter já sido associado com este processo, uma análise detalhada do papel funcional desempenhado por miRNAs durante o dano cardíaco por isquemia-reperfusão não foi ainda efectuada.
    
O principal objectivo deste projecto consiste na identificação de miRNAs capazes de proteger cardiomiócitos humanos contra o dano induzido por isquemia-reperfusão, e caracterizar os mecanismos moleculares subjacentes à sua acção, bem como avaliar o seu potencial terapêutico.

A identificação sistemática e imparcial de miRNA com ac;\ao protectora contra morte celular induzida por isquemia-reperfusão será conseguida através da realização de screenings funcionais de larga escala (high-throughput screening) usando bibliotecas de miRNAs, em células humanas. A identificação e caracterização dos alvos moleculares dos miRNAs selecionados será baseada na combinação de abordagens computacionais e experimentais. A modulação de miRNAs, ou dos seus alvos moleculares, poderá constituir uma abordagem promissora para o desenvolvimento de novas terapias para a patologia cardíaca associada ao dano por isquemia-reperfusão.

Publicizing Sheet


 

The Functional Genomics and RNA-based Therapeutics laboratory welcomes collaborative research projects, both national and international, exploiting genome-wide siRNA and microRNA screenings using high-throughput and high-content screening technologies.

Selected Publications

“Mechanical cues control mutant p53 stability through a Mevalonate/RhoA axis” (2018). E. Ingallina, G. Sorrentino, K. Lisek, R. Bertolio, A. Zannini, L. Azzolin, F. Mantovani, M. Mano, A. Rosato, S. Bicciato, S. Piccolo and G. Del Sal. Nature Cell Biology 20:28-35.

“Id genes are essential for early heart formation” (2017). T. Cunningham, M.S. Yu, W.L. McKeithan, S. Spiering, F. Carrette, C.-T. Huang, P.J. Bushway, M. Tierney, S. Albini, M. Giacca, M. Mano, P.L. Puri, A. Sacco, P. Ruiz-Lozano, J.-F. Riou, M. Umbhauer, G. Duester, M. Mercola and A.R. Colas. Genes and Development 31:1325-1338.

“Analysis of host microRNA function in Shigella infection reveals a role for miR-29b-2-5p in bacterial capture by filopodia” (2017). U. Sunkavalli, C. Aguilar, R.J. Silva, M. Sharan, A.R. Cruz, C. Tawk, C. Maudet, M. Mano and A. Eulalio. PLOS Pathogens 13:e1006327.

“Glucocorticoid receptor signalling controls YAP in breast cancer” (2017). G. Sorrentino, N. Ruggeri, A. Zannini, E. Ingallina, R. Bertolio, C. Marotta, C. Neri, M. Forcato, A. Rosato, M. Mano, S. Bicciato, E. Cappuzzello and G. Del Sal. Nature Communications 8:14073.

"Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction" (2015). M. Mano, R. Ippodrino, S. Zacchigna, L. Zentilin and M. Giacca. Proceedings of the National Academy of Sciences USA 112:11276-11281.

“Functional high-throughput screening identifies the miR-15 microRNA family as cellular restriction factors for Salmonella infection” (2014). C. Maudet, M. Mano, U. Sunkavalli, M. Sharan, M. Giacca, K. Forstner and A. Eulalio. Nature Communications, 5:4718.

“Metabolic control of YAP/TAZ by the Mevalonate Pathway” (2014). G. Sorrentino, N. Ruggeri, V. Specchia, M. Cordenonsi, M. Mano, S. Dupont, A. Manfrin, E. Ingallina, R. Sommaggio, S. Piazza, A. Rosato, S. Piccolo and G. Del Sal. Nature Cell Biology 16:357-366.

“Functional screening identifies microRNAs inducing cardiac regeneration” (2012). A. Eulalio, M. Mano, M. Dal Ferro, L. Zentilin, G. Sinagra, S. Zacchigna and M. Giacca. Nature 492:376-81.

“A Pin1/mutant p53 axis promotes aggressiveness in breast cancer” (2011). J.E. Girardini, M. Napoli, S. Piazza, A. Rustighi, C. Marotta, E. Radaelli, V. Capaci, L. Jordan, P. Quinlan, A. Thompson, M. Mano, A. Rosato, T. Crook, E. Scanziani, A.R. Means, G. Lozano, C. Schneider and G. Del Sal. Cancer Cell 20:79-91.

Patents

“MicroRNA hsa-miR-665 in cardiac hypertrophy” / EPO17160768.2, priority date 14/03/2017

“MicroRNAs for cardiac regeneration through induction of cardiac myocyte proliferation” / PCT/US2012/057590, priority date 23/12/2011

Curriculum Vitae

Career History:
2018-present: Principal Investigator, CNC, Coimbra, Portugal
2014-2018: Assistant Investigator (FCT Investigator Programme), CNC, Coimbra, Portugal
2011-2015: Staff Research Scientist, International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy
2008-2015: Scientific Manager High-Throughput Screening Facility, International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy
2006-2008: Postdoctoral Fellow, Molecular Medicine laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy
2001-2006: Ph.D. in Biochemistry, University of Coimbra, Coimbra, Portugal
1995-2000: Degree in Biochemistry, University of Coimbra, Coimbra, Portugal

Fellowships:
2006-2008: Postdoctoral Fellowship – Portuguese Science Foundation, FCT, Portugal
2001-2006: Ph.D. Fellowship – Portuguese Science Foundation, FCT, Portugal

Prizes:
2014: Starting Grant – FCT Investigator Programme, Portuguese Science Foundation, FCT, Portugal
2011: “Excellence in Research Award”, American Society for Gene and Cell Therapy (ASGCT), USA
2000: “Alvaro J.S. Basto Award” for best final classification in Degree in Biochemistry, University of Coimbra, Portugal