): The primary objective of this proposal is to complete a 3 component career development plan that will result in the applicant's acquiring the necessary skills and experience to develop an independent translational research program investigating the genetic mechanisms responsible for malignant transformation in children. The 3 components of this plan are: 1) a one-year co-mentorship with two senior cancer researcher scientists who will provide him with the guidance and expertise required to complete the academic and research portion of his training: 2) a didactic program that focuses on specific areas of cancer biology; and 3) a transition segment from colleague/mentor to independent research scientist and collaborator during the independent research portion of his proposal. The principal research objectives of this proposal are to investigate the molecular mechanisms and biologic consequences of in vivo somatic genetic events responsible for pediatric malignancies. The hypothesis of the applicants is that somatic mutational events in children with cancer will occur at a higher frequency and with a unique mutational spectra compared to a normal population.
The specific aims which will test the proposed hypotheses are: 1) to determine the frequency and mutational spectra of background and t h erapy induced in vivo somatic mutations in children with specific malignancies (acute lymphocytic leukemia [ALL], Hodgkin's disease, neuroblastoma, and sarcomas) and where possible, to correlate these molecular events with subsequent diseases: 2) to determine if """"""""illegitimate"""""""" V(D)J recombinase mediated mutations occur at a known cancer gene, p53; and 3) to isolate and identify T-lymphocyte imitator phenotype clones from children with relapsed ALL who have an extremely high frequency of background somatic mutations. The frequency of somatic mutational events will be determined by the hprt T-cell cloning assay. Mutational spectra of hprt and p53 mutations will be determined by a variety of methods including multiplex PCR, RT-PCR, IPCR, Southern blotting and DNA sequencing. Clonality of hypermutable clones will be determined by RFLP analysis of TCR gamma and CDR3 region DNA sequence analysis of TCR-beta. These studies will provide the first data on the frequency and mutational spectrum of in vivo spontaneous or treatment induced somatic mutations in children with specific malignancies. The investigators will also begin to determine the potential mutagenic effects associated [with] background """"""""illegitimate"""""""" V(D)J recombinase mediated events in a known cancer gene, p53. In addition, the isolation of mutator phenotype clones will allow for future m e chanistic studies characterizing the genetic and biologic defect(s) associated with leukemogenesis in children.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Scientist Development Award - Research & Training (K01)
Project #
5K01CA077737-04
Application #
6376745
Study Section
Subcommittee G - Education (NCI)
Program Officer
Eckstein, David J
Project Start
1998-09-30
Project End
2002-09-29
Budget Start
2001-09-30
Budget End
2002-09-29
Support Year
4
Fiscal Year
2001
Total Cost
$161,244
Indirect Cost
Name
University of Vermont & St Agric College
Department
Pediatrics
Type
Schools of Medicine
DUNS #
066811191
City
Burlington
State
VT
Country
United States
Zip Code
05405
Murray, Janet M; Messier, Terri; Rivers, Jami et al. (2012) VDJ recombinase-mediated TCR ? locus gene usage and coding joint processing in peripheral T cells during perinatal and pediatric development. J Immunol 189:2356-64
Pinsonneault, Robert L; Vacek, Pamela M; O'Neill, J Patrick et al. (2007) Induction of V(D)J-mediated recombination of an extrachromosomal substrate following exposure to DNA-damaging agents. Environ Mol Mutagen 48:440-50
Finette, Barry A (2006) Analysis of mutagenic V(D)J recombinase mediated mutations at the HPRT locus as an in vivo model for studying rearrangements with leukemogenic potential in children. DNA Repair (Amst) 5:1049-64
Kendall, Heather E; Vacek, Pamela M; Rivers, Jami L et al. (2006) Analysis of genetic alterations and clonal proliferation in children treated for acute lymphocytic leukemia. Cancer Res 66:8455-61
Murray, Janet M; O'Neill, J Patrick; Messier, Terri et al. (2006) V(D)J recombinase-mediated processing of coding junctions at cryptic recombination signal sequences in peripheral T cells during human development. J Immunol 177:5393-404
Rice, Sederick C; Vacek, Pamela; Homans, Alan H et al. (2004) Genotoxicity of therapeutic intervention in children with acute lymphocytic leukemia. Cancer Res 64:4464-71
Kendall, Heather E; Vacek, Pamela M; Finette, Barry A (2004) Analysis of microsatellite instability in children treated for acute lymphocytic leukemia with elevated HPRT mutant frequencies. Mutagenesis 19:409-12
Rice, Sederick C; Vacek, Pamela M; Homans, Alan H et al. (2003) Comparative analysis of HPRT mutant frequency in children with cancer. Environ Mol Mutagen 42:44-9
Finette, Barry A; Kendall, Heather; Vacek, Pamela M (2002) Mutational spectral analysis at the HPRT locus in healthy children. Mutat Res 505:27-41
Yoshioka, M; O'Neill, J P; Vacek, P M et al. (2001) Gestational age and gender-specific in utero V(D)J recombinase-mediated deletions. Cancer Res 61:3432-8

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