# BIOCHEMISTRY OF HYPERTHERMIC CELLULAR INACTIVATION

> **NIH NIH R01** · VANDERBILT UNIVERSITY · 1989 · $119,984

## Abstract

The long term objective is to provide a basic understanding of
hyperthermia in order to increase its therapeutic effectiveness.
This investigation focuses on how changes in the glutathione
concentration, the oxidation of GSH to GSSG and the oxidation of
protein thiols effect thermal sensitivity as well as thermal
enhancement of radiation damage.  The specific aims are:  (1) To
determine if depletion of intracellular glutathione modifies
protein thiol/disulfide (SH/S-S) ratios during hyperthermia thereby
enchancing thermal sensitivity.  Changes in the number of sodium
borohydride reducible protein thiols and protein mixed disulfides
will be correlated to alterations in cell survival.  (2) To determine
the role of  thiol oxidation in the regulation of intracellular
calcium homeostasis during hyperthermia.  Calcium levels,
measured spectrophotometrically using Arsenazo III, will be
determined as a function of GSH level, GSH oxidation, and protein
thiol oxidation.  (3) To determine if thermal sensitivity and
thermal enhancement of radiation damage are affected  by
glutathione depletion during reoxygenation after acute chronic
hypoxia.  Heating plus or minus BSO will occur as a function of
time after reoxgenation and cell survival determined.  In some
experiments, the cells will be irradiated while  hypoxic and then
heated after reoxygenation.  (4) To determine why depletion of
GSH increases thermal sensitivity in CHO cells but not in mouse
LM cells.  The thermal sensitivity of the 2 cell lines measured in
the presence of a GSH depleting agent will be contrasted with
regard to duration of exposure and the mitochondrial GSH
concentration.  (5) To determine if GSH depletion increases the
thermal sensitivity of cells derived from a patient with 5-
oxoprolinuria.  The thermal sensitivity of the GSH mutant will be
measured in the presence and absence of BSO and the results
compared to those obtained from wild type cells.  Changes in cell
survival will be correlated with changes in protein thiols,
mitochondrial GSH levels and calcium levels.  (6) To determine if
thiol oxidation triggers the synthesis of heat shock proteins and/or
the development of thermotolerance.  Heat shock proteins (HSP)
will be measured by gel electrophoresis.  (7) To determine
whether depletion of GSH increases thermal enhancement of
radiation damage by inhibiting repair.  Single strand DNA breaks
will be measured using alkali unwinding and the results correlated
to changes in cell survival.

## Key facts

- **NIH application ID:** 3176105
- **Project number:** 5R01CA038079-06
- **Recipient organization:** VANDERBILT UNIVERSITY
- **SAM.gov UEI:** GTNBNWXJ12D5
- **Principal Investigator:** MICHAEL L. FREEMAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1989
- **Award amount:** $119,984
- **Award type:** 5
- **Project period:** 1984-07-01 → 1991-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3176105

## Citation

> US National Institutes of Health, RePORTER application 3176105, BIOCHEMISTRY OF HYPERTHERMIC CELLULAR INACTIVATION (5R01CA038079-06). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3176105. Licensed CC0.

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