Anuj Mankad

Publications (2)15.58 Total impact

• Article: Natural gene therapy in monozygotic twins with Fanconi anemia.

  Anuj Mankad, Toshiyasu Taniguchi, Barbara Cox, Yassmine Akkari, R Keaney Rathbun, Lora Lucas, Grover Bagby, Susan Olson, Alan D'Andrea, Markus Grompe
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  ABSTRACT: Monozygotic twin sisters, with nonhematologic symptoms of Fanconi anemia (FA), were discovered to be somatic mosaics for mutations in the FANCA gene. Skin fibroblasts, but not lymphocytes or committed hematopoietic progenitors, were sensitive to DNA cross-linking agents. Molecular analysis revealed, in skin cells of both twins, a frameshift causing deletion in exon 27 (2555deltaT) and an exon 28 missense mutation (2670G>A/R880Q). The latter resulted in primarily cytoplasmic expression and reduced function of the mutant FANCA (R880Q) protein. Surprisingly, the same acquired exon 30 missense change (2927G>A/E966K) was detected in the hematopoietic cells of both sisters, but not in their fibroblasts, nor in either parent. This compensatory mutation existed in cis with the maternal exon 28 mutation, and it restored function and nuclear localization of the resulting protein. Both sisters have been free of hematologic symptoms for more than 2 decades, suggesting that this de novo mutation occurred prenatally in a single hematopoietic stem cell (HSC) in one twin and that descendants of this functionally corrected HSC, via intra-uterine circulation, repopulated the blood lineages of both sisters. This finding suggests that treating FA patients with gene therapy might require transduction of only a few hematopoietic stem cells.
  Blood 04/2006; 107(8):3084-90. · 9.90 Impact Factor
• Article: Molecular and functional analysis of SUMF1 mutations in multiple sulfatase deficiency

  Maria Pia Cosma, Stefano Pepe, Giancarlo Parenti, Carmine Settembre, Ida Annunziata, Richard Wade-Martins, Carmela Di Domenico, Paola Di Natale, Anuj Mankad, Barbara Cox, Graziella Uziel, Grazia M.S. Mancini, Enrico Zammarchi, Maria Alice Donati, Wim J. Kleijer, Mirella Filocamo, Romeo Carrozzo, Massimo Carella, Andrea Ballabio
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  ABSTRACT: Multiple sulfatase deficiency (MSD) is a rare disorder characterized by impaired activity of all known sulfatases. The gene mutated in this disease is SUMF1, which encodes a protein involved in a post-translational modification at the catalytic site of all sulfatases that is necessary for their function. SUMF1 strongly enhances the activity of sulfatases when coexpressed with sulfatase in Cos-7 cells. We performed a mutational analysis of SUMF1 in 20 MSD patients of different ethnic origin. The clinical presentation of these patients was variable, ranging from severe neonatal forms to mild phenotypes showing mild neurological involvement. A total of 22 SUMF1 mutations were identified, including missense, nonsense, microdeletion, and splicing mutations. We expressed all missense mutations in culture to study their ability to enhance the activity of sulfatases. Of the predicted amino acid changes, 11 (p.R349W, p.R224W, p.L20F, p.A348P, p.S155P, p.C218Y, p.N259I, p.A279V, p.R349Q, p.C336R, p.A177P) resulted in severely impaired sulfatase-enhancing activity. Two (p.R345C and p.P266L) showed a high residual activity on some, but not all, of the nine sulfatases tested, suggesting that some SUMF1 mutations may have variable effects on the activity of each sulfatase. This study compares, for the first time, clinical, biochemical, and molecular data in MSD patients. Our results show lack of a direct correlation between the type of molecular defect and the severity of phenotype. Hum Mutat 23:576–581, 2004. © 2004 Wiley-Liss, Inc.
  Human Mutation 05/2004; 23(6):576 - 581. · 5.69 Impact Factor