Colgate University junior

Courtney Bellomo is working with Dr. Sarah Anzick on AIB1, a gene amplified in breast and ovarian cancer. The AIB1 protein encodes a protein of 1420 amino acids (Anzick, 1997) and is extremely homologous with TIF2 and SRC-1 (Anzick, 1997). AIB1 is believed to be a coactivator in the signal transduction pathway of estrogen, and for this reason, we feel that that is why overexpression is possibly localized to breast and ovarian cancer. In order to fully understand the role that AIB1 plays in tumor development, we have created a transgene that is a chimera of both the human and mouse homologue of AIB1. This transgene was inserted into embryonic stem cells, and after recombination was determined to occur, the cell was placed into an impregnated female mouse. The mouse containing the transgene has been bred to give F1 mice that are heterozygous for the desired gene. The goal is to cross two heterozygous mice and ultimately, in the F2 generation, breed a homozygous mouse that contains the transgene. In this homozygous mouse, we hope to induce tumors to see if AIB1 is overexpressed. Another aspect of our research is to study the protein expression in the tissues of these mice. Currently, we are in the process of performing Western blots to see the level of expression of AIB1. RNA expression is also being quantitated in order to see if the transcriptional activity has increased. The transgene was also inserted into just the mammary gland of a female mouse in order to study its expression because under the promoter in the transgene construct, the gene should be expressed in day 17 of lactation; that is continuing to be measured. Further studies will also include creating a knock-out construct to see how homozygous mice react.

University of Puerto Rico Cayey senior

Jesus Buonomo is working with Dr. Settara Chandrasekharappa on the isolation and characterization of the promoter region of Zebra-Fish homolog of a Jagged Gene. The aim of the research project is to study the Jagged 1 mutation in humans, and use these fish as a study model.

Virginia Tech junior

Allison Clark is working in the laboratory of Dr. Douglas Wilkin on rapid identification of COL2A1 mutations in Stickler syndrome. Stickler syndrome is one of the milder phenotypes resulting from alterations in COL2A1, the gene which encodes Type II collagen. Type II collagen is the predominant protein in articular cartilage and alterations in this gene can result in skeletal and oral-facial manifestations, as well as vision and hearing defects. COL2A1 has 10 in-frame CGA codons which can mutate to TGA STOP codons via a methylation-deamination mechanism. We have analyzed these sites in genomic DNA from a panel of 40 patients to test the hypothesis that mutations that produce Stickler syndrome preferentially occur at these sites. PCR amplification of genomic DNA containing one of the in-frame CGA codons was done by one of two methods: either using primers which amplify DNA that includes the CGA codon, or using allele specific primers which amplify either normal sequence containing a CGA codon, or amplify a mutant sequence containing a TGA codon. Analysis of PCR products by restriction endonuclease digestion or sequence analysis demonstrated the presence of either normal sequence or mutated sequence. TGA mutations were identified in seven patients, at five of the 10 in-frame CGA codons. One alteration was identified in three unrelated individuals. The identification of these mutations in 7 of 40 probands demonstrates that these sites are common sites for alterations in individuals with Stickler syndrome, and indicates a first step in the search for alterations which result in this disorder.

Washington University freshman

Irena Glick is working with Dr. Doug Wilkin in the Medical Genetics Branch. Irena is helping to identify sites on the COL2A1 gene, which mutate to cause Stickler syndrome. Stickler syndrome is one of the milder human skeletal disorders, but still has many debilitating effects. She amplifies regions of the COL2A1 gene by performing the Polymerase Chain Reaction (PCR) procedure and then uses specific restriction enzymes to cleave the PCR products at the mutation sites. Finally, she analyzes the resulting products by gel electrophoresis. Irena is hoping that a significant number of Stickler syndrome families will have a mutation at the same location on the COL2A1 gene. If this is the case, Irena will assist in developing a simple test for determining whether individuals are genetically inclined towards Stickler syndrome.

University of North Texas graduate student

Judy Hawkins is working with Ann Smith in the Medical Genetics Branch. The aim of the research project is to develop a NIH protocol for evaluating patients with Nail-Patella Syndrome. Judy is also gaining field experience in genetic counseling this summer as a clinical intern at NHGRI. A graduate of Stephen F. Austin State University, Judy is currently a graduate student at the University of North Texas pursuing a Masters degree in biology with an emphasis in genetic counseling. She plans to complete her degree in May of 1999.

University of South Alabama senior

Jennifer Kearley is working with Dr. Anthony Wynshaw-Boris on the effects of oxidative stress on the neurological genetic disorder ataxia-telangiectasia. Ataxia-telangiectasia (AT) is a rare, genetic neurological disorder phenotypically characterized by neurodegeneration, premature aging, radiation sensitivity, and a predisposition to cancer. The primary postulate of this experiment is that the AT disorder results from unchecked oxidative stress, or an accumulation of harmful oxygen free radicals. The chemical Tempol is a drug which is administered to radiation therapy patients to decrease oxidative stress resulting from treatment. In an attempt to correlate oxidative stress to the AT disorder, the drug has been administered to a mouse model genetically altered to simulate humans with the disorder. The goal is to find that the drug increases the life span of these mice and minimizes the neurological degeneration which patients experience. The results so far have indicated that Tempol increases the life span of the mice at least 2-4 months, with the average mouse living for about 2 years. The neurological assays are currently being performed to assess the extent to which neurological degeneration is caused by the event of unchecked oxidative stress.

Northwestern University senior

David Morse is working in the laboratory of Dr. Michael Bittner on the cDNA microarray project. cDNA microarrays are dense, regular deposits of specific DNAs on a glass matrix. The DNA deposits each serve as a hybridization based detector for fluorescently-labeled probe mixtures. The chief use of microarrays is to examine the relative level of gene transcripts when fluorescent representations of the mRNA pools of two different cell lines or tissues are simultaneously hybridized to the array. Fluorescent-labeling allows the amount of hybridization for a particular gene to be detected at each target, and a ratio of the intensities of hybridization for each gene to be calculated. By choosing an appropriate line or tissue as a baseline "normal" level of expression, one can chart which genes have increased or decreased expression in the "abnormal" line or tissue. Alternatively, one can follow changes in gene expression along some time course, such as development, or response to drugs or injury. While there are many current and potential uses, the cDNA Microarray is a developing technology and, like any technology, needs to be scrutinized and improved so the promise this technology holds comes to bear.

Magna Vista High School senior

Emanuel Mullins is working with Dr. Kate Berg on investigating the genetic basis of the disproportionately high prevalence of prostate cancer among African-American men. In collaboration with Howard University, he is also working in a laboratory with Dr. Rick Kittles. In addition to the prostate cancer project, Mullins is also analyzing the historical antecedents of low African-American participation in medical research.

George Washington University senior

Larry Overton works with Christiane Robbins in the Laboratory of Cancer Genetics. Recently a defined region on chromosome 1 by linkage analysis which contains the gene for hereditary prostate cancer (HPC) was found (SCIENCE 274:1371-1374, 1996). A complete physical map of this region was constructed in yeast artificial chromosome (YACs) and bacterial artificial chromosome (BACs). BACs were targeted for sequencing in an effort to identify candidate genes in the linked interval. BACs were subcloned into the M13 bacteriophage vector and PT7 blue plasmid vector, the average insert size being 1kb. DNA was prepared using the Qiagen 9600 BioRobot. A protocol has been designed for extremely high throughput shotgun sequencing of these M13 libraries utilizing the Qiagen 9600 BioRobot and ABI 377 automated fluorescent sequencers. He estimates that ~280 kilobases of raw sequence per day can be generated using this protocol. The data is BLAST searched against all known DNA sequences in the database to find known or homologous gene sequences. In addition, the PHRED/PHRAP algorithms are used to assemble large contigs (several of which are greater than 30 kb) and exon predictions are made. The eventual goal is to identify all genes contained within the several megabase interval known to harbor the HPC gene.

Thomas Wootton High School senior

Josh Richards is working with Dr. Michael Bittner on the Microarray project. As part of the exciting race to map the human genome, DNA microarray technology allows scientists to analyze thousands of genes at the same time. Using this technology, Josh studies a set of genes classified as human-verified genes which have been sequenced by other scientists working at various other genome projects. Microarray involves a complex set of steps to quantify gene expression. The first step in this process includes isolating the DNA from a cell by adding chemicals which lyse the cell. These chemicals then latch onto substances in the cell and trap them when passed through a membrane filter allowing the DNA pass through freely. Next, this filtered DNA is amplified using a process called Polymerase Chain Reaction (PCR). These PCR products are then quantified using a double stranded DNA specific fluorophore before being arrayed (printed).

University of Chicago junior

Lubov Romantseva is working with Dr. Danilo Tagle on a mouse model for Huntington's disease (HD).  HD is a hereditary brain disorder resulting from deterioration and death of brain cells or neurons. Clinical symptoms include increased involuntary motions, loss of control over voluntary motions, and emotional changes such as mood swings and depression. HD can be inherited from either parent who carries the affected gene regardless of the manifestation of the disease. Currently HD is incurable and death occurs usually within 10-15 yrs after onset which most often occurs in midlife. In 1993, the gene for Huntington's disease and the mutation (expansion of the triplet repeat CAG) therein was identified. In the laboratory, Romantseva is involved in monitoring animals for behavioral abnormalities and other phenotypic changes associated with HD. She also tests the mice for the transgene with the use of molecular biology techniques such as DNA extraction and Southern blotting. Other questions which are currently under investigation with the use of the HD mouse model are whether specific genetic alterations that   inhibit one type of apoptosis is able to delay or prevent cell death in neurons of HD mice, and consequently delay onset of HD in these animals. Currently, Romantseva's results show that the delay of HD onset in this mouse model can be used to identify additional targets in finding a potential cure or treatment for HD.

University of Maryland senior

Lao Saal is working with Dr. Khan researching rhabdomyosarcoma (an aggressive, soft tissue tumor of striated muscle with poor prognosis) and developing a genetic fingerprint for rhabdos and other cancers including neuroblastoma, and Ewing's sarcoma. Lao is performing various laboratory techniques, such as: transfecting human cell lines with new genetic information, growing human cancer and normal cells in tissue culture, freezing cell lines, extracting RNA, quantifying and verifying the RNA, constructing fluorescent-labeled cDNA probes from the RNA, hybridizing the cDNA probes onto array slides ("chips") printed with gene clones, and scanning with the microarray confocal laser microscope to see which genes are expressed in various cell lines and controls. Additionally, gene clone sequences and DNA from plasmid clones will be (re)sequenced and verified against the dBEST database. Finally, the data will be analyzed using various statistical and graphical custom and commercial applications and papers for publication will be written.

Bethesda Chevy Chase High School senior

Andrea Seltzer is working with Barbara Biesecker in the Medical Genetics Branch. She is gaining valuable exposure to the field of genetic counseling. Specifically, she has been working on designing a disability course for future graduate students.

Rochester Institute of Technology senior

Jessica Sparrin is working with Barbara Biesecker in the Medical Genetics Branch. She is focusing on the psychosocial aspects of BRCA 1/2 testing of hereditary breast and ovarian cancer families.

Penn State University junior

Michael Stitzel is working in the laboratory of Dr. Lawrence Brody on a potential animal model for neural tube defects. The aim of the research project is to investigate the role of methionine synthase, a gene involved in folic acid metabolism, and to test the effects of its absence on the development of a mouse. Stitzel is measuring the pattern of expression of this gene in developing mouse embryos. This research project may help us better understand the genetic and biochemical basis of neural tube defects such as spina bifida, a disease that affects 1 in 1,000 births. It may cause paralysis of the legs, bladder and bowel complications, recurrent infections, and hydrocephalus (fluid on the brain).

University of Pittsburgh junior

Patricia Stoltzfus is working in the laboratory of Dr. Thomas Ried on the Cancer Chromosome Aberration Project. The aim of the research project is to create both a cytogenetic and physical map of human chromosomes and their genes.The information that the Cancer Chromosome Aberration Project provides will be useful in identifying the exact location of breaks in DNA of the cancer cells. This will improve the diagnosis of cancers and may help to find genes which cause cancer. The research project will create a repository of genetically and physically mapped DNA clones that can be accessed by scientists throughout the country. The clones will be used to detect DNA aberrations with high resolution. The project involves isolation of DNA from bacterial artificial chromosome clones that will be used as probes, or chromosomal markers. Fluorescence in situ hybridization (FISH) is then used to determine the chromosomal map position of all clones. Once cytogeneticists have identified a chromosomal breakpoint with conventional techniques, e.g. chromosome banding of karyotyping, the DNA probes can be used to specifically pinpoint the abnormal chromosome region.

Walt Whitman High School junior

Loyrirk (Float) Temiyakarn is working in the laboratory of Dr. Joan Bailey-Wilson on computer simulation examining Type I and Type II errors in linkage analysis. The aim of the research project is to use computer simulation methods to simulate samples from human populations to determine the effect of failures of assumptions on various methods of human linkage analysis. This may include examining the effect of using incorrect marker allele frequencies and/or ignoring important environmental risk factors in the analyses. The computer program "GASP" will be used to simulate samples of pedigrees. On each individual in the sample, data will be simulated for both a trait and several marker loci. The model used to simulate the trait and markers represents biological truth. These simulated data will be analyzed using several different linkage analysis methods. These analyses will be repeated using both the correct "biological truth" model and also using incorrect models. The results obtained from the analyses will be compared to determine whether the statistical tests are robust and powerful when their assumptions are not met.

Western Maryland College graduate

Maddelena Tilli is working with Dr. Larry Brody and Dr. Ronit Yarden on the interaction of the BRCT domain of BRCA-1 tumor suppressor gene with the Rb-Binding Protein RbAp46. The aim of the research project is to clone gene fragments in a study of the interaction of the BRCT domain of BRCA1 with a Rb binding protein. Mutations of the tumor suppressor gene BRCA1 have been shown to confer an increased susceptibility to both breast and ovarian cancer. One of the highly conserved regions of BRCA1, the acidic carboxy terminal BRCT domain, has been associated with the activation of transcription and may be involved in maintaining genome integrity in other proteins. Tilli currently tries to identify proteins that may interact with the BRCT domain in order to further understand the function of this region in BRCA1. We currently know that BRCA1 interacts with RbAp46, an Rb binding protein. Members of the subfamily in which RbAp46 belongs have been shown to be involved in modifications of histone. Possible involvement of BRCA1 in histone modifications is currently being studied.

Indiana University senior

Jason Wilder is working with Dr. Francis Collins developing a candidate gene database for the FUSION project. FUSION is an international study to find susceptibility genes for non-insulin dependent diabetes. Wilder is working in the laboratory of Dr. Francis Collins on FUSION. The aim of the research project of the FUSION study is to positionally clone susceptibility genes for type II diabetes and diabetes-related quantitative traits.