Alessandra Pelegrini

From Laboratório de Reparo de DNA

Diferença entre revisões)
Revisão de 15:54, 25 Novembro 2011
Root (Discussão | contribs)

← Ver a alteração anterior
Revisão atual
Root (Discussão | contribs)

Linha 1: Linha 1:
{| {|
|- |-
-|[[Image:Alepelegrini.jpg|150px]]+|[[Image:AlePelegrini_Profile.jpg|330px]]
-|Possui bacharelado e licenciatura em Ciências Biológicas pela Universidade Federal do Rio Grande do Sul (2005 e 2006 respectivamente) e mestrado em Biologia Celular e Molecular pela mesma instituição (2007). Está desenvolvendo o projeto de doutorado intitulado "O silenciamento da cinase humana Nek1 altera o reparo normal do DNA" sob orientação do professor Guido Lenz pelo Programa de Pós-Graduação em Biologia Celular e Molecular na Universidade Federal do Rio Grande do Sul. Atualmente realiza estágio de doutoramento no Laboratório de Reparo de DNA, no Instituto de Ciências Biomédicas da Universidade de São Paulo, sob supervisão do professor Carlos Frederico Martins Menck.+|BSc, <b>Biological Sciences</b>, <b>Universidade Federal do Rio Grande do Sul – UFRGS</b>,(2001/2005).<br>MSc, <b>Celular and Molecular Biology</b>, <b>Universidade Federal do Rio Grande do Sul – UFRGS</b>, (2006/2007).<br>PhD, <b>Celular and Molecular Biology</b>, <b>Universidade Federal do Rio Grande do Sul - UFRGS and Universidade de São Paulo – USP</b>, ( 2007/2012).<br>Post-doctoral Fellowship, <b> Cancer Research Center of Toulouse – CRCT</b> (2016/2018)<br>Post-doctoral Fellowship in DNA Repair Lab - <b>Institute of Biological Sciences at University of São Paulo - USP</b> (2013 – Current)<br><br>The researcher has experience in cell and molecular biology, culture of human and animal cells, proteomics and mass spectrometry, microscopy and immunofluorescence, drug testing and cytotoxicity<br><br> Research keywords: <b>Cancer</b>, <b>DNA repair</b>, <b>Replicative stress</b>, <b>Genetic instability</b> and <b>Chemotherapy and Therapeutic resistance</b><br> [http://lattes.cnpq.br/4410437371278362 Lattes].
-<br> +<b>Contato:</b> aleepelegrini[at]gmail.com
- +
|- |-
|colspan=2| |colspan=2|
-== Resumo do Projeto ==+== Current Project ==
-===O silenciamento da cinase humana Nek1 altera o reparo normal do DNA===+Cancer is currently the leading cause of morbidity and mortality worldwide which it is characterized by abnormal and uncontrolled cell growth in addition to genetic instability. It is very likely that genetic instability accelerates and even possibly initiates the proliferation of cancer cells by favoring the emergence of variant cells. Indeed, alterations of genes involved in genome maintenance promote carcinogenesis. This process is predominantly generated in S phase, during the DNA synthesis and relies on a replication stress activated by oncogenes. During the replication, the fork can be stalled for several reasons, such as DNA damage, structured DNA, nucleotide depletion and encounters with the transcription machinery, characterizing replication stress. In this way, replication stress contributes to chromosomal instability since stalled forks may eventually collapse, producing a broken DNA end. Up until recently, the mechanisms leading to fork collapse were only described after prolonged exposition of DNA replication stress sources (24 hours) by Mus81 activity. However, we have been described a phenomenon in which DNA replication forks stalls in response to DNA replication stress and are rapidly converted into DSBs, which we refer to as rapid replication fork breakage (RRFB) and it is dependent of XPF and Artemis nucleases. In this project, i have investigated the role of that endonuclease and its partner, ERCC1, in interstrand crosslink (ICLs) repair and its evolvement in chemotherapy sensitivity. Since the ICLs may cause persistent fork stalling, the proposed objective for this research is understanding how the XPF-ERCC1 complex and each of these separate protein participate in DNA replication stress, exploring their roles from the regulation of fork collapse to repair mechanisms, and evaluating the importance of these early events in the development of genetic instability from chromosomal aberrations.
- +
-A proteína humana Nek1 pertence a uma família de serina/treonina quinases caracterizadas pelo seu envolvimento no ciclo celular que estão presentes em diferentes organismos. Nos últimos anos, cada vez mais estudos estão se focando nessa família de proteínas devido sua relação com patologias do desenvolvimento e à suscetibilidade a tumores. A Nek1, por exemplo, parece estar envolvida na etiologia de duas síndromes hereditárias, a Doença Policística do Rim e a Síndrome de costela curta e polidactilia tipo Majewski. Entretanto, as bases moleculares que relacionam a deficiência dessa proteína a essas patologias não estão estabelecidas. Por isso, alguns estudos buscam entender a função da Nek1 e os achados mais promissores destacam seu papel na ciliogênesis e na resposta ao estresse genotóxico, sugerindo a Nek1 como uma molécula sensora de danos ao material genético e até mesmo participando ativamente de vias de reparo. +
-<br>+
-Em nosso trabalho, através do estudo de células silenciadas por RNA de interferência (figura 1), encontramos um retardo no reparo do DNA em células renais humanas quando tratadas com os agentes genotóxicos metil-metanosulfonato, peróxido de hidrogênio e cisplatina. Quando silenciadas para Nek1, as células não conseguem reparar principalmente as lesões do tipo crosslinks causadas pela cisplatina, como demonstrado através do ensaio cometa nas versões alcalina e neutra (Figura 2). Em vista disso, o objetivo desse projeto é investigar o papel da Nek1 em resposta a lesões causada pelo quimioterápico cisplatina, um indutor de pontes inter e intracadeia de DNA (crosslinks), tentando estabelecer a via de atuação dessa proteína.+
- +
<br> <br>
<center> <center>
 +[[Image:AlePelegriniEsquema.jpg|700px]]
<br> <br>
-[[Image:Alepelegrinifigura1.jpg|350px]] 
<br> <br>
-western-blot demonstrando o silenciamento de Nek1 por shRNA nas linhagens trabalhadas 
-<br><br> 
-[[Image:Alepelegrinifigura2.jpg|350px]] 
-<br> 
-Legenda da figura 2: O Silenciamento da Nek1 altera o reparo normal do DNA de células expostas a Cisplatina. Células Hek293T Wild-type (WT) e silenciadas para Nek1 (KD) foram tratadas com cisplatina por 1h e após diferentes tempos de recuperação o cometa alcalino ou neutro foi realizado conforme descrito em Pelegrini et al., 2010. 
-<br><br> 
</center> </center>
-==Publicações==+== Articles Published in Scientific Journals ==
-Pelegrini, A. L. ; Moura, D. J. ; Brenner, B. L. ; Ledur, P. F. ; Maques, G. P. ; Henriques, J. A. P. ; Saffi, J. ; Lenz, G. (2010). Nek1 silencing slows down DNA repair and blocks DNA damage-induced cell cycle arrest. Mutagenesis, 25,447-454.+Melo-Hanchuk TD, Slepick PF, Meirelles GV, Basei FL, Lovato DV, Granato DC, Pauletti BA, Domingues RR, Leme AFP, <b>Pelegrini AL</b>, Lenz G, Knapp S, Elkins JM and Kobarg J. NEK1 kinase domain structure and its dynamic protein interactome after exposure to Cisplatin. <u>Scientific Reports</u> 2017; 7: (5445) 1-13.<br>[https://www.nature.com/articles/s41598-017-05325-w Full Text]<br><br>
-Gerhardt, D; Horn, A. P. ; Gaelzer, M. M. ; Frozza, R. L. ; Delgado-Cañedo, A. ; Pelegrini, A. L. ; Henriques, Amélia T. ; Lenz, G. ; Salbego, C. (2009). Boldine: a potential new antiproliferative drug against glioma cell lines. Investigational New Drugs, 27, 517-525.+Filippi-Chiela EC, Thomé MP, Bueno e Silva MM, <b>Pelegrini AL</b>, Ledur PF, Garicochea B, Zamin LL, Lenz G. Resveratrol abrogates the temozolomide-induced G2 arrest leading to mitotic catastrophe and reinforces the temozolomide-induced senescence in glioma cells. <u>BMC Cancer</u> 2013; 13: 147.<br> [https://bmccancer.biomedcentral.com/articles/10.1186/1471-2407-13-147 Full Text]<br><br>
-</center>+Lopez PL, Filippi-Chiela EC, Silva AO, Cordero EA, Garcia-Santos D, <b>Pelegrini AL</b>, Reder GM, Barbieri NL, Lenz G. Sensitization of glioma cells by x-linked inhibitor of apoptosis protein knockdown. <u>Oncology</u> 2012; 83: 75-82.<br>[https://www.karger.com/Article/FullText/337978 Full Text]<br><br>
 +<b>Pelegrini AL</b>, Moura DJ, Brenner BL, Ledur PF, Maques GP, Henriques JA, Saffi J, Lenz G. Nek1 silencing slows down DNA repair and blocks DNA damage-induced cell cycle arrest. <u>Mutagenesis</u> 2010; 25: 447-54.<br>[https://academic.oup.com/mutage/article-lookup/doi/10.1093/mutage/geq026 Full Text]<br><br>
 +Gerhardt D, Horn AP, Gaelzer MM, Frozza RL, Delgado-Cañedo A, <b>Pelegrini AL</b>, Henriques AT, Lenz G, Salbego C. Boldine: a potential new antiproliferative drug against glioma cell lines. <u>Invest New Drugs</u> 2009; 27, 517-25.<br> [https://dx.doi.org/10.1007/s10637-008-9203-7 Full Text]<br><br>

Revisão atual

BSc, Biological Sciences, Universidade Federal do Rio Grande do Sul – UFRGS,(2001/2005).
MSc, Celular and Molecular Biology, Universidade Federal do Rio Grande do Sul – UFRGS, (2006/2007).
PhD, Celular and Molecular Biology, Universidade Federal do Rio Grande do Sul - UFRGS and Universidade de São Paulo – USP, ( 2007/2012).
Post-doctoral Fellowship, Cancer Research Center of Toulouse – CRCT (2016/2018)
Post-doctoral Fellowship in DNA Repair Lab - Institute of Biological Sciences at University of São Paulo - USP (2013 – Current)

The researcher has experience in cell and molecular biology, culture of human and animal cells, proteomics and mass spectrometry, microscopy and immunofluorescence, drug testing and cytotoxicity

Research keywords: Cancer, DNA repair, Replicative stress, Genetic instability and Chemotherapy and Therapeutic resistance
Lattes.

Contato: aleepelegrini[at]gmail.com


Current Project

Cancer is currently the leading cause of morbidity and mortality worldwide which it is characterized by abnormal and uncontrolled cell growth in addition to genetic instability. It is very likely that genetic instability accelerates and even possibly initiates the proliferation of cancer cells by favoring the emergence of variant cells. Indeed, alterations of genes involved in genome maintenance promote carcinogenesis. This process is predominantly generated in S phase, during the DNA synthesis and relies on a replication stress activated by oncogenes. During the replication, the fork can be stalled for several reasons, such as DNA damage, structured DNA, nucleotide depletion and encounters with the transcription machinery, characterizing replication stress. In this way, replication stress contributes to chromosomal instability since stalled forks may eventually collapse, producing a broken DNA end. Up until recently, the mechanisms leading to fork collapse were only described after prolonged exposition of DNA replication stress sources (24 hours) by Mus81 activity. However, we have been described a phenomenon in which DNA replication forks stalls in response to DNA replication stress and are rapidly converted into DSBs, which we refer to as rapid replication fork breakage (RRFB) and it is dependent of XPF and Artemis nucleases. In this project, i have investigated the role of that endonuclease and its partner, ERCC1, in interstrand crosslink (ICLs) repair and its evolvement in chemotherapy sensitivity. Since the ICLs may cause persistent fork stalling, the proposed objective for this research is understanding how the XPF-ERCC1 complex and each of these separate protein participate in DNA replication stress, exploring their roles from the regulation of fork collapse to repair mechanisms, and evaluating the importance of these early events in the development of genetic instability from chromosomal aberrations.



Articles Published in Scientific Journals

Melo-Hanchuk TD, Slepick PF, Meirelles GV, Basei FL, Lovato DV, Granato DC, Pauletti BA, Domingues RR, Leme AFP, Pelegrini AL, Lenz G, Knapp S, Elkins JM and Kobarg J. NEK1 kinase domain structure and its dynamic protein interactome after exposure to Cisplatin. Scientific Reports 2017; 7: (5445) 1-13.
Full Text

Filippi-Chiela EC, Thomé MP, Bueno e Silva MM, Pelegrini AL, Ledur PF, Garicochea B, Zamin LL, Lenz G. Resveratrol abrogates the temozolomide-induced G2 arrest leading to mitotic catastrophe and reinforces the temozolomide-induced senescence in glioma cells. BMC Cancer 2013; 13: 147.
Full Text

Lopez PL, Filippi-Chiela EC, Silva AO, Cordero EA, Garcia-Santos D, Pelegrini AL, Reder GM, Barbieri NL, Lenz G. Sensitization of glioma cells by x-linked inhibitor of apoptosis protein knockdown. Oncology 2012; 83: 75-82.
Full Text

Pelegrini AL, Moura DJ, Brenner BL, Ledur PF, Maques GP, Henriques JA, Saffi J, Lenz G. Nek1 silencing slows down DNA repair and blocks DNA damage-induced cell cycle arrest. Mutagenesis 2010; 25: 447-54.
Full Text

Gerhardt D, Horn AP, Gaelzer MM, Frozza RL, Delgado-Cañedo A, Pelegrini AL, Henriques AT, Lenz G, Salbego C. Boldine: a potential new antiproliferative drug against glioma cell lines. Invest New Drugs 2009; 27, 517-25.
Full Text

Ferramentas pessoais