Rose-Mary Na'Aman Boustany, M.D.

Adjunct Professor in the Department of Pediatrics

Department:
Pediatrics

Division:
Neurology

Email:
boust001mcdukeedu

Mailing Address:
MSRB Box 2604, Room 285
Durham, NC 27710

Telephone:
919-684-3219

Training:
M.D., American University of Beirut School of Medicine

Last Updated:
March 08, 2004

Research Interests:
Overview:
Our Laboratory is investigating the presence and mechanism(s) of apoptosis operative in Batten disease and other neurodegenerative diseases. So far, lipid raft structure, regulation of sphingolipid levels and their trafficking have been implicated as a theme in a number of the neuronal ceroid lipofuscinoses.  We have also identified a novel Batten variant using Affymetrix GeneChip technology, and are zeroing in on the gene defect.

Batten disease is an autosomal recessive neurodegenerative disorder resulting in neuronal and photoreceptor death. We have established the presence of apoptosis in Batten disease. This was done by traditional methods such as TUNEL and PI staining, FACS analysis and electron microscopy, and by analyzing apoptosis-related genes that are dysregulated by genechip analysis.

CLN3-deficient juvenile variant: We determined that normally the CLN3 protein is made in the Golgi and travels via Rab4 and Rab11-positive recycling endosomes to lipid rafts in the plasma membrane. CLN3 harbors at its carboxyl end a number of conserved amino acids and potential glycosylation sites that are necessary for preserving antiapoptotic function and normal growth of cells. Defining these regions of the gene becomes important when one considers the facts that CLN3 is overexpressed in a number of cancers, and blocking its expression by antisense strategies results in cancer cell growth inhibition, as well as apoptotic death. These include a VYFAE motif similar to the VFFAE motif in the ß amyloid precursor mutated in some cases of familial Alzheimer disease.  This motif is embedded in a structurally defined galactosylceramide lipid raft binding domain common to CLN3 protein, ß amyloid precursor, infectious prionic protein, and the V3 loop of the gp120 protein of HIV. We have shown that CLN3 protein and galactosylceramide co-localize to Golgi and in lipid rafts.
Mutant CLN3 protein in patient cells lacks this structural motif, is trapped in a fragmented Golgi, is mis-directed to Rab7-positive endosomes, and never appears in recycling endosomes or the plasma membrane.  Moreover, CLN3 deficient cells have increased levels of ceramide, sphingomyelin, globoside, glucosylceramide, and galactosylceramide. Neither CLN3 nor galactosylceramide are found in lipid rafts in CLN3deficient cells. Transfection of CLN3-deficient cells with intact CLN3 cDNA restores the presence of CLN3 and galactosylceramide both to lipid rafts, and corrects ceramide levels to normal.
CLN6-deficient variant: We are in the process of defining subcellular localization of another, recently identified Batten gene called CLN6 that seems to complement CLN3-deficient cells with respect to both growth and apoptotic defects.
CLN9-deficient variant: Another novel Batten variant we call CLN9 was identified by scrutinizing genechip profiles of Batten disease cell lines.  CLN9-deficient cells manifest rapid growth, increased apoptosis, a cell adhesion defect and abnormal levels of a number of complex sphingolipids including sphingomyelin, glucosylceramide, ceramide trihexoside and globoside, as well as ceramide. CLN8, the gene defective in Northern Epilepsy with Mental Retardation, complements CLN9-deficient cells with respect to growth and apoptotic defects, and partially corrects sphingomyelin levels. This suggests that CLN8 and CLN9 interact at a functional level.  There is evidence that CLN3, CLN6 and CLN8 also complement each other functionally, raising the question of whether all these proteins form functional complexes.

This work will provide a handle to unravel key information pertinent to protein and lipid trafficking within the cell, and the pathobiology of these neurodegenerative diseases and may provide answers that will lead to intelligent and targeted therapies for these and other neurodegenerative diseases as well as cancer.

Publications:
2002 -- Pubmed # 11830536 -- Rylova SN, Amalfitano A, Persaud-Sawin DA, Guo WX, Chang J, Jansen PJ, Proia AD, Boustany RM. The CLN3 gene is a novel molecular target for cancer drug discovery. Cancer Res. 2002 Feb 1;62(3):801-8.

2002 -- Pubmed # 11921051 -- Dhar S, Bitting RL, Rylova SN, Jansen PJ, Lockhart E, Koeberl DD, Amalfitano A, Boustany RM. Flupirtine blocks apoptosis in batten patient lymphoblasts and in human postmitotic CLN3- and CLN2-deficient neurons. Ann Neurol. 2002 Apr;51(4):448-66.

2002 -- Pubmed # 12123853 -- Lockhart EM, Warner DS, Pearlstein RD, Penning DH, Mehrabani S, Boustany RM. Allopregnanolone attenuates N-methyl-D-aspartate-induced excitotoxicity and apoptosis in the human NT2 cell line in culture. Neurosci Lett. 2002 Aug 2;328(1):33-6.

2002 -- Pubmed # 11791207 -- Gao H, Boustany RM, Espinola JA, Cotman SL, Srinidhi L, Antonellis KA, Gillis T, Qin X, Liu S, Donahue LR, Bronson RT, Faust JR, Stout D, Haines JL, Lerner TJ, MacDonald ME. Mutations in a novel CLN6-encoded transmembrane protein cause variant neuronal ceroid lipofuscinosis in man and mouse. Am J Hum Genet. 2002 Feb;70(2):324-35.

2002 -- Pubmed # 12189165 -- Persaud-Sawin DA, VanDongen A, Boustany RM. Motifs within the CLN3 protein: modulation of cell growth rates and apoptosis. Hum Mol Genet. 2002 Sep 1;11(18):2129-42.

2001 -- Pubmed # 11354831 -- Ashley-Koch A, Bonner ER, Gaskell PC, West SG, Tim R, Wolpert CM, Jones R, Farrell CD, Nance M, Svenson IK, Marchuk DA, Boustany RM, Vance JM, Scott WK, Pericak-Vance MA. Fine mapping and genetic heterogeneity in the pure form of autosomal dominant familial spastic paraplegia. Neurogenetics. 2001 Mar;3(2):91-7.

2001 -- Pubmed # 11309678 -- Svenson IK, Ashley-Koch AE, Gaskell PC, Riney TJ, Cumming WJ, Kingston HM, Hogan EL, Boustany RM, Vance JM, Nance MA, Pericak-Vance MA, Marchuk DA. Identification and expression analysis of spastin gene mutations in hereditary spastic paraplegia. Am J Hum Genet. 2001 May;68(5):1077-85.

2000 -- Pubmed # 11087788 -- Hentati A, Deng HX, Zhai H, Chen W, Yang Y, Hung WY, Azim AC, Bohlega S, Tandan R, Warner C, Laing NG, Cambi F, Mitsumoto H, Roos RP, Boustany RM, Ben Hamida M, Hentati F, Siddique T. Novel mutations in spastin gene and absence of correlation with age at onset of symptoms. Neurology. 2000 Nov 14;55(9):1388-90.

1997 -- Puranam K, Qian W-H, Nikbakht K, Venable M, Obeid L, Hannun Y, and Boustany R-M:  Upregulation of Bcl-2 and Elevation of Ceramide in Batten Disease.  Neuropediatrics 1997; 28:1-5.

1997 -- Bennett MJ, Boriak RL, and Boustany R-M.  Polyunsaturated fatty acids reverse the lysosomal storage and accumulation of subunit 9 of mitochondrial F1F0-ATP synthase in cultured lymphoblasts from patients with Batten Disease.  J. Inher. Metab. Dis., 1997, 20:457-460.

1997 -- Munroe PB, Mitchison HM, O"Rawe AM, Anderson JW, Boustany R-M, Lerner TJ, Taschner PEM, de Vos Nanneke, Breuning MH, Gardiner RM, and Mole SE.  Spectrum of Mutation in the Batten Disease Gene, CLN3.  Am. J. of Hum. Genetics, 1997,  61:310-316.

1997 -- Scott WK, Gaskell PC, Lennon F, Wolpert C, Arnold MM, Aylsworth AS, Warner C, Farrell CD, Boustany R-M, Albright SG, Boyd E, Kingston HM, Cumming WJK, Vance JM, and Pericak-Vance MA.  Locus heterogeneity, anticipation and reduction of the chromosome 2p minimal candidate region in autosomal dominant familial spastic paraplegia.  Neurogenetics, 1997, 195-202.

1995 -- Johnson, DW, Speier S, Qian WH, Lane SC, Cook A, Suzuki K, Daniel P, Boustany R-M:  Role of Subunit-9 of mitochondrial ATP synthase in Batten Disease.  Am. J. Med. Genet. 1995; 57:350-360.