# MOLECULAR ANALYSIS OF GENETIC MUTATION IN CHILDREN

> **NIH NIH K11** · UNIVERSITY OF VERMONT & ST AGRIC COLLEGE · 1994 · $94,608

## Abstract

The aim of this research is to explore the molecular mechanisms of
somatic gene mutations in the pediatric population, and to examine the
implications of these in vivo events on human development, tumorigenesis,
secondary malignancies, and the monitoring of autoimmune diseases.  In
Phase I, the focus is to determine the somatic mutant frequency in the
hprt gene of normal human pediatric subjects from birth to late
adolescents.  Analysis of the molecular rearrangements in hprt negative
clones, specifically the frequency of V(D)J recombinase mediated events
resulting in the deletion of exons 2 and 3 will be performed.  In Phase
II, the aim is to examine hprt somatic mutations in children prior to,
during, and after the completion of chemotherapy and/or radiation
treatment for a malignant process.  The frequency of thioguanine
resistant T-cell clones will be determined and compared with normal
subjects.  Folic acid levels will be correlated with mutant frequency and
a master bank set up for long term follow up of these patients for
determining if the frequency and types of somatic gene rearrangements can
be used as markers for predicting secondary malignancies.  The second
hypothesis in Phase II will examine the treatment of pre, and new onset
diabetic children with azathioprine utilizing the principle of surrogate
selection to isolate hprt negative T-cell clones.  The frequency of
thioguanine resistant T-cell lymphocyte clones will be determined prior
to, during and after the completion of azathioprine treatment.  Analysis
of molecular rearrangements as well as the clonality of these hprt
negative clones will be determined.

The techniques to be utilized include: 1) T-cell cloning assay, selecting
for thioguanine resistant clones. 2) Southern blot analysis of mutant and
wildtype clones to determine TCR clonality and structural changes at the
hprt locus. 3) Multiplex PCR analysis of hprt gene for the detection of
deletions and point mutations. 4) DNA sequence analysis of hprt mutant
clones to determine V(D)J recombinase mediated events.

These investigations should answer questions on the importance of somatic
mutations in developing healthy children and those children with cancer.
It will also provide the foundation for devising a novel approach to
monitoring azathioprine treatment of Type I diabetes.

## Key facts

- **NIH application ID:** 2194546
- **Project number:** 5K11HD001017-02
- **Recipient organization:** UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
- **SAM.gov UEI:** Z94KLERAG5V9
- **Principal Investigator:** BARRY Alan FINETTE
- **Activity code:** K11 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1994
- **Award amount:** $94,608
- **Award type:** 5
- **Project period:** 1993-07-01 → 1997-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2194546

## Citation

> US National Institutes of Health, RePORTER application 2194546, MOLECULAR ANALYSIS OF GENETIC MUTATION IN CHILDREN (5K11HD001017-02). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2194546. Licensed CC0.

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