Normand Beaulieu
University of Toledo
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Publication
Featured researches published by Normand Beaulieu.
Nature Genetics | 2003
Marie-France Robert; Steves Morin; Normand Beaulieu; Ian Chute; Annie Barsalou; A. Robert MacLeod
Transcriptional silencing by CpG island methylation is a prevalent mechanism of tumor-suppressor gene suppression in cancers. Genetic experiments have defined the importance of the DNA methyltransferase Dnmt1 for the maintenance of methylation in mouse cells and its role in neoplasia. In human bladder cancer cells, selective depletion of DNMT1 with antisense inhibitors has been shown to induce demethylation and reactivation of the silenced tumor-suppressor gene CDKN2A. In contrast, targeted disruption of DNMT1 alleles in HCT116 human colon cancer cells produced clones that retained CpG island methylation and associated tumor-suppressor gene silencing, whereas HCT116 clones with inactivation of both DNMT1 and DNMT3B showed much lower levels of DNA methylation, suggesting that the two enzymes are highly cooperative. We used a combination of genetic (antisense and siRNA) and pharmacologic (5-aza-2′-deoxycytidine) inhibitors of DNA methyl transferases to study the contribution of the DNMT isotypes to cancer-cell methylation. Selective depletion of DNMT1 using either antisense or siRNA resulted in lower cellular maintenance methyltransferase activity, global and gene-specific demethylation and re-expression of tumor-suppressor genes in human cancer cells. Specific depletion of DNMT1 but not DNMT3A or DNMT3B markedly potentiated the ability of 5-aza-2′-deoxycytidine to reactivate silenced tumor-suppressor genes, indicating that inhibition of DNMT1 function is the principal means by which 5-aza-2′-deoxycytidine reactivates genes. These results indicate that DNMT1 is necessary and sufficient to maintain global methylation and aberrant CpG island methylation in human cancer cells.
Journal of Biological Chemistry | 2002
Normand Beaulieu; Steves Morin; Ian Chute; Marie-France Robert; Hannah Nguyen; A. Robert MacLeod
Abnormal methylation and associated silencing of tumor suppressor genes is a common feature of many types of cancers. The observation of persistent methylation in human cancer cells lacking the maintenance methyltransferase DNMT1 suggests the involvement of other DNA methyltransferases in gene silencing in cancer. To test this hypothesis, we have evaluated methylation and gene expression in cancer cells specifically depleted of DNMT3A or DNMT3B,de novo methyltransferases that are expressed in adult tissues. Here we have shown that depletion of DNMT3B, but not DNMT3A, induced apoptosis of human cancer cells but not normal cells. DNMT3B depletion reactivated methylation-silenced gene expression but did not induce global or juxtacentromeric satellite demethylation as did specific depletion of DNMT1. Furthermore, the effect of DNMT3B depletion was rescued by exogenous expression of either of the splice variants DNMT3B2 or DNMT3B3 but not DNMT1. These results indicate that DNMT3B has significant site selectivity that is distinct from DNMT1, regulates aberrant gene silencing, and is essential for cancer cell survival.
Journal of Biological Chemistry | 1999
Fournel M; Sapieha P; Normand Beaulieu; Jeffrey M. Besterman; MacLeod Ar
A common event in the development of human neoplasia is the loss of growth regulatory tumor suppressor functions. Methylation of 5′ CpG islands of tumor suppressor genes and elevated levels of the DNA-(cytosine-5)-methyltransferase enzyme (DNA MeTase) are also prevalent features of human neoplasia. However, direct evidence that elevated DNA MeTase levels alter gene expression and influence oncogenesis has been difficult to obtain, in part due to the lack of specific DNA MeTase inhibitors. Here we show that specific reduction of cellular DNA MeTase levels in human cancer cells with potent antisense inhibitors: 1) causes demethylation of the p16 ink4A gene promoter; 2) causes re-expression of the p16ink4A protein; 3) leads to accumulation of the hypophosphorylated form of the retinoblastoma protein (pRb); and 4) inhibits cell proliferation. Stepwise reduction of cellular DNA MeTase protein levels also induced a corresponding rapid increase in the cell cycle regulator p21WAF/Cip1 protein demonstrating a regulatory link between DNA MeTase and the growth regulator p21WAF/Cip1 that is independent of methylation of DNA. These results suggest that the elevated levels of DNA MeTase seen in cancer cells can inhibit tumor suppressors by distinct mechanisms involving either transcriptional inactivation through DNA methylation or by a methylation independent regulation.
Cancer Research | 2006
Frederic J. Reu; Douglas W. Leaman; Ratan R. Maitra; Soo In Bae; Leonid Cherkassky; Mark W. Fox; Donald R. Rempinski; Normand Beaulieu; A. Robert MacLeod; Ernest C. Borden
Resistance of human renal cell carcinoma (RCC) and melanoma to the apoptosis-inducing effects of IFNs was postulated to result from epigenetic silencing of genes by DNA methylation, a common feature of human cancers. To reverse silencing, 5-AZA-deoxycytidine (5-AZA-dC) or selective depletion of DNA methyltransferase 1 (DNMT1) by phosphorothioate oligonucleotide antisense (DNMT1 AS) were employed in cells resistant (<5% terminal deoxynucleotidyl transferase-mediated nick-end labeling positive) to apoptosis induction by IFN-alpha2 and IFN-beta (ACHN, SK-RC-45, and A375). 5-AZA-dC and DNMT1 AS similarly depleted available DNMT1 protein and, at doses that did not cause apoptosis alone, resulted in apoptotic response to IFNs. The proapoptotic tumor suppressor RASSF1A was reactivated by DNMT1 inhibitors in all three cell lines. This was associated with demethylation of its promoter region. IFNs augmented RASSF1A protein expression after reactivation by DNMT1 inhibition. In IFN-sensitive WM9 melanoma cells, expression of RASSF1A was constitutive but also augmented by IFNs. RASSF1A small interfering RNA reduced IFN-induced apoptosis in WM9 cells and in DNMT1-depleted ACHN cells. Conversely, lentiviral expression of RASSF1A but not transduction with empty virus enabled IFN-induced apoptosis. IFN induced tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and TRAIL-neutralizing antibody inhibited apoptotic response to IFN in RASSF1A-expressing ACHN cells. Accordingly, RASSF1A markedly sensitized to recombinant TRAIL. Normal kidney epithelial cells, although expressing RASSF1A, did not undergo apoptosis in response to IFN or TRAIL but had >400-fold higher TRAIL decoy receptor 1 expression than transduced ACHN cells (real-time reverse transcription-PCR). Results identified RASSF1A as regulated by IFNs and participating in IFN-induced apoptosis at least in part by sensitization to TRAIL.
Bioorganic & Medicinal Chemistry Letters | 2008
Stephen William Claridge; Franck Raeppel; Marie-Claude Granger; Naomy Bernstein; Oscar Mario Saavedra; Lijie Zhan; David Llewellyn; Amal Wahhab; Robert Deziel; Jubrail Rahil; Normand Beaulieu; Hannah Nguyen; Isabelle Dupont; Annie Barsalou; Carole Beaulieu; Ian Chute; Serge Gravel; Marie-France Robert; Sylvain Lefebvre; Marja Dubay; Roussen Pascal; Jeff Gillespie; Zhiyun Jin; James C. Wang; Jeffrey M. Besterman; A. Robert MacLeod; Arkadii Vaisburg
A series of thieno[3,2-b]pyridine-based inhibitors of c-Met and VEGFR2 tyrosine kinases is described. The compounds demonstrated potency with IC(50) values in the low nanomolar range in vitro while the lead compound also showed in vivo activity against various human tumor xenograft models in mice. Further exploration of this class of compounds is underway.
Bioorganic & Medicinal Chemistry Letters | 2009
Michael Mannion; Stephane Raeppel; Stephen William Claridge; Nancy Zhou; Oscar Mario Saavedra; Ljubomir Isakovic; Lijie Zhan; Frédéric Gaudette; Franck Raeppel; Robert Deziel; Normand Beaulieu; Hannah Nguyen; Ian Chute; Carole Beaulieu; Isabelle Dupont; Marie-France Robert; Sylvain Lefebvre; Marja Dubay; Jubrail Rahil; James C. Wang; Hélène Ste-Croix; A. Robert MacLeod; Jeffrey M. Besterman; Arkadii Vaisburg
A series of N-(4-(6,7-disubstituted-quinolin-4-yloxy)-3-fluorophenyl)-2-oxo-3-phenylimidazolidine-1-carboxamides targeting c-Met and VEGFR2 tyrosine kinases was designed and synthesized. The compounds were potent against these two enzymes with IC(50) values in the low nanomolar range in vitro, possessed favorable pharmacokinetic profiles and showed high efficacy in vivo in several human tumor xenograft models in mice.
Bioorganic & Medicinal Chemistry Letters | 2009
Stephane Raeppel; Stephen William Claridge; Oscar Mario Saavedra; Frédéric Gaudette; Lijie Zhan; Michael Mannion; Nancy Zhou; Franck Raeppel; Marie-Claude Granger; Ljubomir Isakovic; Robert Deziel; Hannah Nguyen; Normand Beaulieu; Carole Beaulieu; Isabelle Dupont; Marie-France Robert; Sylvain Lefebvre; Marja Dubay; Jubrail Rahil; James C. Wang; Hélène Ste-Croix; A. Robert MacLeod; Jeffrey M. Besterman; Arkadii Vaisburg
A series of N-(3-fluoro-4-(2-arylthieno[3,2-b]pyridin-7-yloxy)phenyl)-2-oxo-3-phenylimidazolidine-1-carboxamides targeting c-Met and VEGFR2 tyrosine kinases was designed and synthesized. The compounds were potent against these two enzymes with IC(50) values in the low nanomolar range in vitro, possessed favorable pharmacokinetic profiles and showed high efficacy in vivo in several human tumor xenograft models in mice.
Bioorganic & Medicinal Chemistry Letters | 2009
Oscar Mario Saavedra; Stephen William Claridge; Lijie Zhan; Franck Raeppel; Marie-Claude Granger; Stephane Raeppel; Michael Mannion; Frédéric Gaudette; Nancy Zhou; Ljubomir Isakovic; Naomy Bernstein; Robert Deziel; Hannah Nguyen; Normand Beaulieu; Carole Beaulieu; Isabelle Dupont; James C. Wang; A. Robert MacLeod; Jeffrey M. Besterman; Arkadii Vaisburg
A family of thieno[3,2-b]pyridine based small molecule inhibitors of c-Met and VEGFR2 were designed based on lead structure 2. These compounds were shown to have IC(50) values in the low nanomolar range in vitro and were efficacious in human tumor xenograft models in mice in vivo.
Bioorganic & Medicinal Chemistry Letters | 2010
Stephane Raeppel; Frédéric Gaudette; Michael Mannion; Stephen William Claridge; Oscar Mario Saavedra; Ljubomir Isakovic; Robert Deziel; Normand Beaulieu; Carole Beaulieu; Isabelle Dupont; Hannah Nguyen; James C. Wang; A. Robert MacLeod; Christiane R. Maroun; Jeffrey M. Besterman; Arkadii Vaisburg
A novel series of N-(3-fluoro-4-(2-substituted-thieno[3,2-b]pyridin-7-yloxy)phenyl)-1-phenyl-5-(trifluoromethyl)-1H-pyrazole-4-carboxamides targeting RON receptor tyrosine kinase was designed and synthesized. SAR study of the series allowed us to identify compounds possessing either inhibitory activity of RON kinase enzyme in the low nanomolar range with low residual activity against the closely related c-Met or potent dual inhibitory activity against RON and c-Met, with no significant activity against VEGFR2 in both cases.
Bioorganic & Medicinal Chemistry Letters | 2010
Frédéric Gaudette; Stephane Raeppel; Hannah Nguyen; Normand Beaulieu; Carole Beaulieu; Isabelle Dupont; A. Robert MacLeod; Jeffrey M. Besterman; Arkadii Vaisburg
A novel series of malonamide-type dual VEGFR2/c-Met inhibitors in which one of the amide bonds was replaced by an amide isostere-a trifluoroethylamine unit, was designed, synthesized, and evaluated for their enzymatic and cellular inhibition of VEGFR2 and c-Met enzymes. Optimization of these molecular entities resulted in identification of potent and selective inhibitors of VEGFR2 enzyme.