User:Orsolya Kiraly

Contact Info

• Research Affiliate, Engelward Laboratory
• Department of Biological Engineering
• Massachusetts Institute of Technology
• 77 Massachusetts Avenue 16-760, Cambridge, MA
• okiraly AT mit DOT edu

Education

• Ph.D., Biochemistry, Semmelweis University, Budapest, Hungary, 2008
• B.S., Biology, ELTE, Budapest, Hungary, 2002
• I was shaped greatly by an education at ELTE Apaczai High School

Research interests

My current work is about genome rearrangements (large-scale mutations) that can lead to cancer

I received my PhD for work on how mutations in pancreatic trypsin inhibitor cause inflammation. In the Engelward lab, my work is aimed at genome rearrangements in the pancreas in vivo.

Genome rearrangements are a hallmark of cancer cells. They can be deletions, inversions or duplications that can drive cancer by activating oncogenes or inactivating tumor suppressor genes. Rearrangements can form by homologous recombination (HR), which is an important DNA repair/tolerance mechanism but can lead to genetic changes.

Using a transgenic reporter mouse, we have found that the formation of HR-driven rearrangements is governed by an interplay of genes, environment and tissue physiology. This is an example of gene-environment interactions and may be used to identify people with a higher risk for cancer. Further, we found that inflammation, a major cancer risk factor, induces HR-driven rearrangements through increasing both DNA damage and regenerative proliferation.

These findings have led to the generation of an advanced reporter mouse for use in ongoing studies on HR-driven genetic changes, and a collaborative project developing mitigators of inflammation-induced tissue injury.

Past projects

Functional effects of sequence variants in promoters, introns, and protein-coding regions

• In the laboratory of Miklos Sahin-Toth at Boston University, I determined the functional effects of mutations in SPINK1, the pancreatic secretory trypsin inhibitor. Inhibition of trypsin activity by SPINK1 is important because trypsin activity can result in the activation of other digestive enzymes in a cascade reaction, which can lead to cell damage and pancreatic inflammation. We found that signal peptide mutations abolish the secretion of SPINK1 into pancreatic juice, and coding region mutations cause misfolding of the protein which is then degraded intracellularly and is not secreted. In patients with these mutations, spontaneously activated trypsin is thus not inhibited by SPINK1, eventually resulting in autodigestion and inflammation.

• As side projects in the Sahin-Toth Lab, I initiated testing the functional effects of intronic mutations in SPINK1, and also tested a patient-derived mutation in cationic trypsinogen. Autoactivation of trypsinogen had been linked to pancreatic inflammation and this mutation had been hypothesized to increase autoactivation. However, testing this hypothesis was challenging because we couldn’t express intact trypsinogen with the mutation. We therefore developed a novel expression system, which has been included in Methods in Molecular Biology.

• At Semmelweis University I was working in a team investigating the D4 dopamine receptor gene. This gene was the first to be investigated in psychiatric genetics association studies, and its variants are associated with certain personality traits and disorders such as ADHD. However, the functional effects of these variants (and thus the molecular basis of phenotypic associations) were not clear. Using a reporter gene assay, we found that a duplication in the promoter decreases transcriptional efficiency, potentially influencing the abundance of receptor molecules and neurotransmission. However, the most widely studied SNP in the gene had no effect on gene expression in our assay. The apparent effect of this SNP in association studies may be due to another variant which is in linkage disequilibrium with the candidate SNP.

• My undergraduate thesis project at the Agricultural Biotechnology Research Center (in Gödöllő, Hungary) was aimed at generating host factor independent mutants of the 16-3 phage integrase by protein engineering. Integrases catalyze site-specific recombination, which is harnessed in gene targeting. The excellent mentoring I received at ABC gave me a strong foundation in the laboratory techniques of molecular biology, and this project raised my interest in DNA metabolism and recombination.

Publications

Research papers

See them on PubMed

Book chapters

Kiraly O, Guan L, Sahin-Toth M. Expression of Recombinant Proteins with Uniform N-Termini. In: Ming-Qun X, Evans T (Eds): Heterologous Protein Expression in E. coli, Methods in Molecular Biology 705, Springer, Berlin, 2011, pp. 175-194.

Nemoda Z, Kiraly O, Barta C, Sasvari-Szekely M. Pharmacogenetic Aspects of Dopaminergic Neurotransmission-Related Gene Polymorphisms. In: Darvas F, Guttman A, Dormán G (Eds): Chemical Genomics, Marcel Dekker Inc., New York, 2003, pp. 275-313.

Bookmarks

Laboratory tools and resources

• Qiagen bench guide

• Centrifuge calculator

• Berglund:PCR_Additives A good list of PCR additives

• GeneCards

• Information Hyperlinked over Proteins

• VassarStats: Online stats tool

Useful links

• Classification of carcinogens according to the International Agency for Research on Cancer

• Environmental Working Group

• The Comparative Toxicogenomics Database

• American Association for Cancer Research

Good books

• The Suffering Gene is a very readable book about the various environmental exposures that can damage DNA

• Cancer, the Evolutionary Legacy describes why we are susceptible to cancer even without environmental exposures

• Correcting the Blueprint of Life: An Historical Account of the Discovery of DNA Repair Mechanisms by Errol C. Friedberg is a fascinating story of scientific discovery, great scientist personalities and the unpredictability of research

• At the End of an Age by historian John Lukacs is not a scientific book, but it contains a deeply informed reflection on the nature of historical and scientific knowledge. Read a shorter essay on this here

• Also by Lukacs, Confessions of an Original Sinner is a rich account of a journey through very different places and times, full of sharp observations

Miscellaneous

• New animal species are being discovered to this day, the latest being the Tarzan chameleon

• Read about which Hungarian or Hungarian-born scientists gave the world the ballpoint pen, holography, Intel, Rubik’s cube, vitamin C, stress, and the nuclear chain reaction