Pain in the Neck: An Investigation of TSHr Gene Expression

1. Pain in the Neck: An Investigation of TSHr Gene Expression in a Population with Abundant Hypothyroidism Wesley Anderson and Ronald Kaltreider, Ph.D. Department of Biological Sciences, York College of Pennsylvania Wesley Anderson in surgery removing a fibroid in Apam, Ghana Materials & Methods An example of a goiter in a subject Table 2. CT of each sample’s grouped replicate with respect to genetic probe used. Sample # 5 Sample #6 Introduction A B C D B C D Extract RNA & convert to cDNA Obtain 30 tumor samples • A population in Apam, Ghana has a high frequency of adult females with goiter formation with significant levels of iodine in the diet. Since 2005, over 200 females have had sub-total thyroidectomies as a result of goiter in this population of 20,000.  • The development of a goiter in humans is frequently attributed to a lack of iodine in the diet, however it can be attributed to a genetic malfunction.  • Thyroid stimulating hormone receptor (TSHR) mutations have previously been shown to cause hereditary goiter formation (Kitanaka et al. 2005).  • A mutation which decreases the activity of the TSHR would cause decreased activity by the thyroid gland. A possible mechanism for compensating for this would be increasing the number of TSHRs on the gland, which would be evidenced by an increase in mRNA coding for the production of TSHR.  • The extraction of mRNA from tumor specimens will allow for conversion to cDNA and a detection of the concentration of TSHR mRNA present. Comparing these levels to a specimen without goiter will determine if TSHR levels are elevated in the population of interest. Design primer for TSH receptor (Akcurin et al. 2007) 500 bp 400 bp TSHr primer 300 bp 18s primer 200 bp 100 bp Unsuccessful banding PCR sample cDNA aMean • Conclusion • Failed to reject H1; when compared to control with Dunn’s Multiple Comparison’s test P>0.05 • Failed to reject H0; 18s was not amplified in each sample so concentration correlation cannot be established • A lack of conclusive data- more samples should be tested Successful banding Fig. 2. A 2% agarose gel with two TSHr primers (lanes B & C) run with two tumor samples’ cDNA against a 100 bp ladder (lane A). Lane D is control 18s gene probe. Run Rt-PCR samples Statistical Analysis • Future Studies • Examine other genetic causes • Record dissolved I2 concentration • Compare chromosome 14 sequencing Results • Works Cited • Akcurin, S., Doga, T., Tysoe, C., Ellard, S., Leener, A., Vassart, G., Costagliola, S. 2008. A family with a novel TSH receptor activating germline mutation. European Journal of Pediatrics. 167:1231-1237. • Kitanaka, S., Takeda, A., Sato, U., Miki, Y., Hishinuma, A., Ieiri, T., Igarashi, T. 2005. A novel compound heterozygous mutation in the thyroglobulin gene resulting in congenital goitrous hypothyroidism with high serum triiodothyronine levels. Journal of Human Genetics. 51:376-382. Hypothesis H1- Individuals will show an increased mRNA levels of TSHr in tumor samples H0- Individuals will have no difference in mRNA levels of TSHr in tumor samples Fig. 3. Amplification Plot of TSHr gene expression in 11 samples plus 2 control samples using Rt-PCR SYBR Green Flourescence Fig. 1. Comparison of number of goiters removed from females from Apam, Ghana of varying age (yrs) by Building Solid Foundations medical team between 2006 and 2009. Acknowledgements We thank Dr. Richard Kurz , Sr for alerting us to this population, Dr. Robert Davis and Dr. Thomas Davis for their instruction and assistance in removal of the tumor specimen, and the people of Apam, Ghana for their generosity and friendliness.