Annual Report 2011 - Center for Advanced Biotechnology and ...

More documents

Recommendations

Info

Dr. James Millonig CABM Resident Faculty Member Associate Professor Department of Neuroscience and Cell Biology UMDNJ-Robert Wood Johnson Medical School Adjunct Assistant Professor Department of Genetics Rutgers University Dr. Paul Matteson Research Associate Anna Dulencin Graduate Fellow Myka Ababon Graduate Student Ji Yeon Choi Graduate Student Silky Kamdar Graduate Student Bo Li Graduate Student Mai Soliman Graduate Student Developmental Neurogenetics Laboratory Dr. James H. Millonig came to CABM in September 1999 from the Rockefeller University where he was a postdoctoral fellow in the laboratory of Dr. Mary E. Hatten. His postdoctoral research combined neurobiology and mouse genetics to characterize and clone the dreher mouse locus. He did his doctoral research at Princeton University with Dr. Shirley M. Tilghman. Dr. Millonig is a recipient of the March of Dimes Basil O’Connor Starter Research Award and grants from the National Institutes of Health, Department of Defense, National Alliance for Research on Schizophrenia and Depression, Autism Speaks, National Alliance for Autism Research, N.J. Governor’s Council on Autism and New Jersey Commission on Spinal Cord Research. He is director of the RU/Princeton/RWJMS M.D./Ph.D. program. Autism Spectrum Disorder (ASD) Individuals diagnosed with ASD exhibit deficiencies in communication and reciprocal social interactions that are accompanied by rigid or repetitive interests and behaviors. ASD is considered to be a neurodevelopmental disorder that has a polygenic basis. Increased risk for the disorder is believed to be due to multiple genes interacting with each other as well as environmental factors. Risk for ASD is likely due to both genetic and non-genetic environmental factors, with epigenetic regulation of genes providing a possible interface between genetic and environmental factors. Our previous research has focused on the homeobox transcription factor, ENGRAILED 2 (EN2). The common alleles (underlined) of two intronic EN2 SNPs, rs1861972 (A/G) and rs1861973 (C/T), are significantly associated with ASD (518 families, P=.00000035). Subsequent association, LD mapping, and re-sequencing identified the associated rs1861972-rs1861973 as the most suitable candidate to test for function. Function was then tested in primary cultures of cerebellar granule cells since endogenous En2 is expressed at high levels in these cells. Luciferase assays demonstrated that the A-C haplotype results in an ~250% increase in expression. Additional analysis determined that both associated alleles are sufficient and necessary for this activator function. EMSAs revealed that nuclear factors specifically bind the A-C haplotype. An unbiased proteomic screen identified two transcription factors, Cux1 and NfiB, that bind significantly better to the A-C haplotype. Knock-down, over-expression, ChIP and EMSA supershift analysis demonstrated that both transcription factors bind the A-C haplotype at the same time and are required for activator function. These studies determined that the ASD-associated A-C haplotype functions as a transcriptional activator. 25
We then investigated if EN2 levels are increased in individuals with autism. Ninety cerebellar samples were acquired and Taqman QRTPCR measured normalized EN2 mRNA levels. A significant increase was observed in affected individuals with an A-C haplotype compared to controls (74% increase, P=0.0005). Thus the A-C haplotype is also correlated with increased EN2 levels in individuals with ASD. Because ASD is a neurodevelopmental disorder, we generated transgenic mice. EN2 was replaced with a DsRed, reporter and 25kb of evolutionarily conserved sequence was used to drive expression of the transgene. 6 A-C and 8 G-T lines were generated. QRTPCR measured Ds-Red levels for all the lines at 5 developmental time points (E9.5, E12.5, E17.5, P6 and adult). At all stages the A-C haplotype results in increased expression (P
Page 1: CENTER FOR ADVANCED BIOTECHNOLOGY A
Page 4 and 5: Director’s Overview CABM—25 and
Page 6 and 7: of Health, National Science Foundat
Page 8 and 9: Cell & Developmental Biology Labora
Page 10 and 11: protein metabolism/activity, such t
Page 12 and 13: Dr. Céline Gélinas CABM Resident
Page 14 and 15: (CHINJ, CINJ and CABM) and Lauri Go
Page 16 and 17: Dr. Masayori Inouye CABM Resident F
Page 18 and 19: Xu, Z., Mirochnitchenko, O., Xu, C.
Page 20 and 21: Dr. Fang Liu CABM Resident Faculty
Page 22 and 23: Publications: Matsuura, I., Chiang,
Page 24 and 25: In the prior year, we have made con
Page 28 and 29: We have mapped 5 different vl modif
Page 30 and 31: Dr. Arnold Rabson Director Child He
Page 32 and 33: Publications: Zheng, X., Cui, X.X.,
Page 34 and 35: polymerase II, facilitating the sel
Page 36 and 37: Dr. Mengqing Xiang CABM Resident Fa
Page 38 and 39: neurons diminish while there is a s
Page 40 and 41: Laboratory Faculty Director Structu
Page 42 and 43: Human transcription factors are typ
Page 44 and 45: atomic detail and have yielded nume
Page 46 and 47: Figure from Das et al., 2011 showin
Page 48 and 49: Dr. Helen M. Berman Board of Govern
Page 50 and 51: Dr. Chenghua Shao Research Associat
Page 52 and 53: Dr. Joseph Marcotrigiano CABM Resid
Page 54 and 55: The long-term goals of our studies
Page 56 and 57: s Publications: Singarapu, K.K., Mi
Page 58 and 59: Zhao, L., Zhao, K., Hurst, R., Slat
Page 60 and 61: Acton, T.B., Xiao, R., Anderson, S.
Page 62 and 63: Kryshtafovych, A., Bartual, S.G., B
Page 64 and 65: properties by lowering susceptibili
Page 66 and 67: Dr. Ann M. Stock Associate Director
Page 68 and 69: Publications: Barbieri, C.M., Mack,
Page 70 and 71: Lectures and Seminars 69
Page 72 and 73: CABM RETREAT, June 30, 2011 Cook Ca
Page 75 and 76: 25 th Anniversary CABM Symposium Fr
Page 77 and 78:
Edery Lab Joseph Albistur George Gh
Page 79 and 80:
PATENTS P. Lobel, et al. “Methods
Page 81 and 82:
Montelione, G.T., Ma, L. and Krug,
Page 83 and 84:
Outside Academic Members Dr. Wayne
Page 85 and 86:
Former CABM Scientific Advisory Boa
Page 87 and 88:
Fiscal Information 86
Page 89 and 90:
Anderson, Stephen Rutgers CABM Gran
Page 91 and 92:
Leukemia & Lymphoma Society BFL-1 a
Page 93 and 94:
Total Award Start - End FY 2011 Dir
Page 95 and 96:
Department of Defense Tumor suppres
Page 97 and 98:
Total Award Start - End FY 2011 Dir
show all

Annual Report 2011 - Center for Advanced Biotechnology and ...

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?