Institute for Biogenesis Research
University of Hawaii John A. Burns School of Medicine

Supported by IBR-COBRE (Center of Biomedical Research Excellence) IDeA Program
NCRR 5P20RR024206-01, NIGMS 8P20GM103457-05

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Name Johann Urschitz
Affiliation JABSOM
Position Assistant Professor
Degree Ph.D.
Phone 808-956-7417
Fax 808-956-7316
Email johann@hawaii.edu
Address IBR, 1960 East-West Rd., Biomed E112, Honolulu, HI 96822
 

Research projects:

Development of non-viral transposase-based vectors for gene therapy and transgenesis
Impact of maternal and paternal obesity on adult onset metabolic disease for subsequent generations

Description of research:

Non-viral transposase-based vectors for gene therapy and transgenesis
PiggyBac, an insect transposase is a highly efficient tool for gene transfer in a broad range of organisms. It is able to mediate integration of large gene cassettes of more than 100kb and has been shown to achieve long-term transgene expression in vitro and in vivo and for genome modifications of animals by various assistive reproduction techniques such as ICSI, CTI or PNI. We have improved the plasmid constructs to be self-inactivating and to contain the whole transpositional machinery on a single hyperactive and helper-independent plasmid (pmhyGENIE-3/4) thereby reducing the genotoxic potential while simultaneously improving integration efficiency. Recently, we have used these vectors to (1) address the role of TrkB signaling in malignant transformation, (2) explore pmGENIE-3 plasmids approach for designing DNA vaccines, (3) large animal transgenesis and (4) structure-function analysis of mouse SRY gene.

Impact of maternal and paternal obesity
In recent decades, the increase in obesity, particularly in industrialized countries has taken on epidemic proportions. For example, in the US alone, two-thirds of adults 20 years old and over are overweight or obese (with a body mass index or BMI above 25kg/m2. Should this trend continue, 75% of US adults are estimated to be overweight or obese by 2015. An even higher prevalence of obesity exists for minority populations. Obesity rates for Native Hawaiians and other Pacific Islanders are 2-3 times higher than the general population while African Americans are 1.4 times more likely to be obese than Non-Hispanic Whites. Maternal obesity has been shown to have long-term consequences for offspring health and wellbeing, however molecular mechanisms mediating the effects of obesity and maternal nutrition during pregnancy on fetal growth and health later in life have yet to be fully elucidated. We have developed a strategy for achieving a placenta-specific reduction in Glut1 gene expression using the pmGENIE-3 system. Our intention is to decrease the glucose transport into the placenta in vivo in a maternal adiposity mouse model and evaluate if this approach will attenuate fetal overgrowth and subsequent metabolic syndrome in offspring. Specifically, we will use the pmhyGENIE-3 system to permanently integrate micro-RNA adapted shRNA (shRNAmir) constructs targeting mouse Glut1 into the genome of placental mouse trophoblasts. We will deliver this vector in a tissue specific manner in vivo by utilizing Ultrasound Targeted Microbubble Destruction (UTMD), an innovative approach for gene transfer that combines very high tissue specificity with minimally invasive delivery. In contrast to the attention being devoted to the influence of maternal lifestyle on the long-term health of their offspring, few studies have investigated the potential influence of paternal nutritional status or body composition on the next generation. Male obesity has tripled over the last few decades and adversely impacts reproductive success. As a consequence a majority of male partners seeking assistive reproduction techniques (ART) are overweight or obese. We are interested in examining the extent to which sperm mediates the transmission of environmental information and paternal health traits. We will therefore explore if paternal obesity with or without ICSI induces epigenetic changes in spermatozoa that may promote the inter- or transgenerational inheritance of adult onset diseases.


Selected Publications:

  • Urschitz J, Kawasumi M, Owens J, Morozumi K, Yamashiro H, Stoytchev I, Marh J, Dee JA, Kawamoto K, Coates CJ, Kaminski JM, Pelczar P, Yanagimachi R, Moisyadi S. Helper-independent piggyBac plasmids for gene delivery approaches: Strategies for avoiding potential genotoxic effects. Proc. Natl. Acad. Sci (USA) 2010;107 (18):8117-8122. PMID:20404201
  • Owens JB, Urschitz j, Stoytchev I, Dang NC, Stoytchev Z, Belcaid M, Maragathavally KJ, Coates, Segal DJ, Moisyadi S. Chimeric piggyBac Transposases for Genomic Targeting in Human Cells. Nucleic Acids Res. 2012 Apr 9. Epub. PMID:22492708
  • Marh J, Stoytcheva Z, Urschitz J, Sugawara A, Yamashiro H, Owens H, Stoytchev I, Pelczar P, Yanagimachi R, Moisyadi S. Hyperactive self-inactivating piggyBac for transposase-enhanced pronuclear microinjection transgenesis. Proc. Natl. Acad. Sci (USA) 2012; 109:19184-9.
  • Wu Z, Xu Z, Zou X, Zeng F, Shi J, Liu D, Urschitz J, Moisyad S, Li Z. Pig transgenesis by piggyBac transposition in combination with somatic cell nuclear transfer. Transgenic Res. 2013 Jul 16. [Epub ahead of print]. PMID: 23857557
  • Urschitz J and Moisyad S. Transpositional Transgenesis with piggyBac. Mobile Genetic Elements 3:3, e25167; May 1, 2013.
  • Bertino P, Urschitz J, Hoffmann FW, Youc BR, Rose AH, Park WH, Moisyadi S, and Hoffmann PR. Vaccination with a piggyBac plasmid that directs transgene integration leads to sustained antigen expression and CD8+ T cell responses. (manuscript accepted 01/22/14 by Vaccine).
  • DeWitt J, Ochoa V, Urschitz J, Elston M, Moisyadi S, Nishi R. Constitutively active TrkB confers an aggressive transformed phenotype to a neural crest derived cell line. Oncogene. 2013 Mar 4. doi:10.1038/onc.2013.39.
  • Anderson CD, Urschitz J, Khemmani M, Owens JB, Moiysiadi S, Shohet RV, Walton CB. Ultrasound directs a transposase system for durable hepatic gene delivery in mice. Ultrasound in Medicine and Biology doi:10.1016/j.ultrasmedbio.2013.07.002
  • Stollberg J, Urschitz J, Urban Z, Boyd CD. A quantitative evaluation of SAGE. Genome Res. 2000 Aug;10(8):1241-8. PMID:0958642
  • Urschitz J, Iobst S, Urban Z, Granda C, Souza KA, Lupp C, Schilling K, Scott I, Csiszar K, Boyd CD. A serial analysis of gene expression in sun-damaged human skin. J Invest Dermatol. 2002 Jul;119(1):3-13. PMID:12164917
  • Urschitz, J. Photoaging of Skin: A Functional Genomics Approach. Dissertation, June 2004
  • Brizzolara S, Kileen J, Urschitz J. Gene Expression in Pelvic Organ Prolapse, Mol Hum Reprod. 2009 Jan;15(1):59-67. PMID:19056808