Open Access. Powered by Scholars. Published by Universities.®

Medicine and Health Sciences Commons

Open Access. Powered by Scholars. Published by Universities.®

Articles 1 - 30 of 125

Full-Text Articles in Medicine and Health Sciences

Optimizing Crispr/Cas9 For Gene Silencing Of Sod1 In Mouse Models Of Als, Zachary C. Kennedy Aug 2019

Optimizing Crispr/Cas9 For Gene Silencing Of Sod1 In Mouse Models Of Als, Zachary C. Kennedy

GSBS Dissertations and Theses

Mutations in the SOD1 gene are the best characterized genetic cause of amyotrophic lateral sclerosis (ALS) and account for ~20% of inherited cases and 1-3% of sporadic cases. The gene-editing tool Cas9 can silence mutant genes that cause disease, but effective delivery of CRISPR-Cas9 to the central nervous system (CNS) remains challenging. Here, I developed strategies using canonical Streptococcus pyogenes Cas9 to silence SOD1. In the first strategy, I demonstrate effectiveness of systemic delivery of guide RNA targeting SOD1 to the CNS in a transgenic mouse model expressing human mutant SOD1 and Cas9. Silencing was observed in both the brain ...


National Trends In Elective Ileal Pouch-Anal Anastomosis For Ulcerative Colitis, Chau Maggie Hoang Jun 2018

National Trends In Elective Ileal Pouch-Anal Anastomosis For Ulcerative Colitis, Chau Maggie Hoang

GSBS Dissertations and Theses

Background: Recent national trends and distribution of ileal pouch-anal anastomosis (IPAA) procedures for patients with ulcerative colitis (UC) are unknown. We examined the frequency of use of elective IPAA procedures among patients with UC and the distribution of IPAA procedures across more than 140 U.S. academic medical centers and their affiliates.

Methods: Data were obtained from the University HealthSystem Consortium for patients with a primary diagnosis of UC admitted electively between 2012 and 2015.

Results: The mean age of the study population (n=6,875) was 43 years and 57% were men. Among these, one-third (n=2,307) underwent ...


Engineered Exosomes For Delivery Of Therapeutic Sirnas To Neurons, Reka A. Haraszti May 2018

Engineered Exosomes For Delivery Of Therapeutic Sirnas To Neurons, Reka A. Haraszti

GSBS Dissertations and Theses

Extracellular vesicles (EVs), exosomes and microvesicles, transfer endogenous RNAs between neurons over short and long distances. We have explored EVs for siRNA delivery to brain. (1) We optimized siRNA chemical modifications and siRNA conjugation to lipids for EV-mediated delivery. (2) We developed a GMP-compatible, scalable method to manufacture active EVs in bulk. (3) We characterized lipid and protein content of EVs in detail. (4) We established how protein and lipid composition relates to siRNA delivering activity of EVs, and we reverse engineered natural exosomes (small EVs) into artificial exosomes based on these data.

We established that cholesterol-conjugated siRNAs passively associate ...


A Translational Pathway For Recombinant Adeno-Associated Virus Human Gene Therapy: From Target Identification And Animal Modeling Of The Disease To Non-Human Primate And Human Studies, Alisha Gruntman Nov 2016

A Translational Pathway For Recombinant Adeno-Associated Virus Human Gene Therapy: From Target Identification And Animal Modeling Of The Disease To Non-Human Primate And Human Studies, Alisha Gruntman

GSBS Dissertations and Theses

Many steps go into developing a clinical viral gene therapy. The course starts with appropriate disease selection and moves through the many hurdles of in-vitro testing, animal model validation and proof-of-concept studies, all the way through pre-clinical large animal studies. In this thesis, I propose to outline the process of developing a translation pathway for a gene therapy using recombinant adeno-associated virus (rAAV). I will expand on this outline using data that I have generated during the course of my Ph.D. that ranges from animal model validation all the way through pre-clinical vector stability studies. Two disease models will ...


Targeting Drug Resistance In Chronic Myeloid Leukemia: A Dissertation, Leyuan Ma Nov 2016

Targeting Drug Resistance In Chronic Myeloid Leukemia: A Dissertation, Leyuan Ma

GSBS Dissertations and Theses

Inhibiting BCR-ABL kinase activity with tyrosine kinase inhibitors (TKIs) has been the frontline therapy for CML. Resistance to TKIs frequently occurs, but the mechanisms remain elusive.

First, to uncover survival pathways involved in TKI resistance in CML, I conducted a genome-wide RNAi screen in human CML cells to identify genes governing cellular sensitivity to the first generation TKI called IM (Gleevec). I identified genes converging on and activating the MEK/ERK pathway through transcriptional up-regulation of PRKCH. Combining IM with a MEK inhibitor synergistically kills TKI-resistant CML cells and CML stem cells.

Next, I performed single cell RNA-seq to compare ...


A Walk On The Fine Line Between Reward And Risk: Aav-Ifnβ Gene Therapy For Glioblastoma: A Dissertation, Dwijit Guhasarkar Jul 2016

A Walk On The Fine Line Between Reward And Risk: Aav-Ifnβ Gene Therapy For Glioblastoma: A Dissertation, Dwijit Guhasarkar

GSBS Dissertations and Theses

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor. The current standard-of-care treatment including surgery, radiation and temozolomide (TMZ) chemotherapy does not prolong the survival satisfactorily. Here we have tested the feasibility, efficacy and safety of a potential gene therapy approach using AAV as gene delivery vehicle for treatment of GBM.

Interferon-beta (IFNβ) is a cytokine molecule also having pleiotropic anticancerous properties. Previously it has been shown by our group that AAV mediated local (intracranial) gene delivery of human IFNβ (hIFNβ) could be an effective treatment for non-invasive human glioblastoma (U87) in orthotopic xenograft mouse model.But ...


Barriers And Facilitators To Deaf Trauma Survivors’ Help-Seeking Behavior: Lessons For Behavioral Clinical Trials Research: A Master’S Thesis, Melissa L. Anderson May 2016

Barriers And Facilitators To Deaf Trauma Survivors’ Help-Seeking Behavior: Lessons For Behavioral Clinical Trials Research: A Master’S Thesis, Melissa L. Anderson

GSBS Dissertations and Theses

Deaf individuals experience significant obstacles to participating in behavioral health research when careful consideration is not given to accessibility in the design of study methodology. To inform such considerations, we conducted a secondary analysis of a mixed-methods study that explored 16 Deaf trauma survivors’ help-seeking experiences. Our objective was to identify key findings and qualitative themes from consumers' own words that can be applied to the design of behavioral clinical trials methodology. In many ways, the themes that emerged are what we would expect of any research participant, Deaf or hearing – a need for communication access, empathy, respect, strict confidentiality ...


Developing An Adeno-Associated Viral Vector (Aav) Toolbox For Cns Gene Therapy: A Dissertation, Sourav Roy Choudhury Jan 2016

Developing An Adeno-Associated Viral Vector (Aav) Toolbox For Cns Gene Therapy: A Dissertation, Sourav Roy Choudhury

GSBS Dissertations and Theses

Neurological disorders – disorders of the brain, spine and associated nerves – are a leading contributor to global disease burden with a sizable economic cost. Adeno-associated viral (AAV) vectors have emerged as an effective platform for CNS gene therapy and have shown early promise in clinical trials. These trials involve direct infusion into brain parenchyma, an approach that may be suboptimal for treatment of neurodegenerative disorders, which often involve more than a single structure in the CNS. However, overall neuronal transduction efficiency of vectors derived from naturally occurring AAV capsids after systemic administration is relatively low. We have developed novel capsids AAV-AS ...


Gene Therapy For Amyotrophic Lateral Sclerosis: An Aav Delivered Artifical Microrna Against Human Sod1 Increases Survival And Delays Disease Progression Of The Sod1G93a Mouse Model: A Dissertation, Lorelei I. Stoica Dec 2015

Gene Therapy For Amyotrophic Lateral Sclerosis: An Aav Delivered Artifical Microrna Against Human Sod1 Increases Survival And Delays Disease Progression Of The Sod1G93a Mouse Model: A Dissertation, Lorelei I. Stoica

GSBS Dissertations and Theses

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by loss of motor neurons, resulting in progressive muscle weakness, atrophy, paralysis and death within five years of diagnosis. About ten percent of cases are inherited, of which twenty percent are due to mutations in the superoxide dismutase 1 (SOD1) gene. Since the only FDA approved ALS drug prolongs survival by just a few months, new therapies for this disease are needed. Experiments in transgenic ALS mouse models have shown that decreasing levels of mutant SOD1 protein alters and in some cases entirely prevents disease progression. We explored this potential ...


Gender Differences In Choice Of Procedure And Case Fatality Rate For Elderly Patients With Acute Cholecystitis: A Masters Thesis, Courtney E. Collins Dec 2015

Gender Differences In Choice Of Procedure And Case Fatality Rate For Elderly Patients With Acute Cholecystitis: A Masters Thesis, Courtney E. Collins

GSBS Dissertations and Theses

Background: Treatment decisions for elderly patients with gallbladder pathology are complex. Little is known about what factors go into treatment decisions in this population. We used Medicare data to examine gender-based differences in the use of cholecystectomy vs. cholecystostomy tube placement in elderly patients with acute cholecystitis.

Methods: We queried a 5% random sample of Medicare data (2009-2011) for patients >65 admitted for acute cholecystitis (by ICD-9 code) who subsequently underwent a cholecystectomy and/or cholecystostomy tube placement. Demographic information (age, race), clinical characteristics (Elixhauser index, presence of biliary pathology), and hospital outcomes (case fatality rate, length of stay, need ...


Sentinel Lymph Node Biopsy In Elderly Patients With Intermediate Thickness Melanoma: A Masters Thesis, Kate H. Dinh May 2015

Sentinel Lymph Node Biopsy In Elderly Patients With Intermediate Thickness Melanoma: A Masters Thesis, Kate H. Dinh

GSBS Dissertations and Theses

Background: A landmark study suggested that wide excision of intermediate-thickness melanoma with sentinel lymph node biopsy (SLNB) and subsequent completion lymph node dissection (CLND) for regional disease may improve prognostication and disease-free survival (DFS) compared with those undergoing wide excision alone. However, these benefits were relatively small and not associated with an improvement in disease-specific survival (DSS). It remains unknown if SLNB and subsequent treatments are beneficial in elderly patients who have a decreased overall (OS) due to other causes.

Methods: Adults ≥ 70 years of age, who underwent surgical intervention for intermediate-thickness cutaneous melanoma from 2000-2013 were identified from a ...


Use Of Opioids For Pain Management In Nursing Homes: A Dissertation, Camilla B. Pimentel Apr 2015

Use Of Opioids For Pain Management In Nursing Homes: A Dissertation, Camilla B. Pimentel

GSBS Dissertations and Theses

Nursing homes are an essential yet understudied provider of cancer-related care for those with complex health needs. Nine percent of nursing home residents have a cancer diagnosis at admission, and it is estimated that one-third of them experience pain on a daily basis. Although pain management is an essential component of disease treatment, few studies have evaluated analgesic medication use among adults with cancer in this setting. Use of opioids, which are the mainstay of pain management in older adults because of their effectiveness in controlling moderate to severe pain, may be significantly related to coverage by the Medicare Part ...


Raav-Mediated Gene Transfer For Study Of Pathological Mechanisms And Therapeutic Intervention In Canavan's Disease: A Dissertation, Seemin Seher Ahmed Dec 2014

Raav-Mediated Gene Transfer For Study Of Pathological Mechanisms And Therapeutic Intervention In Canavan's Disease: A Dissertation, Seemin Seher Ahmed

GSBS Dissertations and Theses

Canavan’s Disease is a fatal Central Nervous System disorder caused by genetic defects in the enzyme – aspartoacylase and currently has no effective treatment options. We report additional phenotypes in a stringent preclinical aspartoacylase knockout mouse model. Using this model, we developed a gene therapy strategy with intravenous injections of the aspartoacylase gene packaged in recombinant adeno associated viruses (rAAVs). We first investigated the CNS gene transfer abilities of rAAV vectors that can cross the blood-brain-barrier in neonatal and adult mice and subsequently used different rAAV serotypes such as rAAV9, rAAVrh.8 and rAAVrh.10 for gene replacement therapy. A ...


Psychometric Evaluation Of Joint-Specific Patient-Reported Outcome Measures Before And After Total Knee Replacement: A Dissertation, Barbara L. Gandek Sep 2014

Psychometric Evaluation Of Joint-Specific Patient-Reported Outcome Measures Before And After Total Knee Replacement: A Dissertation, Barbara L. Gandek

GSBS Dissertations and Theses

Background: Patient reports of pain and function are used to inform the need for and timing of total knee replacement (TKR) and evaluate TKR outcomes. This dissertation compared measurement properties of commonly-used patient surveys in TKR and explored ways to develop more efficient knee-specific function measures.

Methods: 1,179 FORCE-TJR patients (mean age=66.1, 61% female) completed questionnaires before and 6 months after TKR. Patient surveys included the knee-specific Knee injury and Osteoarthritis Outcome Score (KOOS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and generic SF-36 Health Survey. Tests of KOOS and WOMAC measurement properties included evaluations ...


Approaches And Considerations Towards A Safe And Effective Adeno-Associated Virus Mediated Therapeutic Intervention For Gm1-Gangliosidosis: A Dissertation, Cara M. Weismann Aug 2014

Approaches And Considerations Towards A Safe And Effective Adeno-Associated Virus Mediated Therapeutic Intervention For Gm1-Gangliosidosis: A Dissertation, Cara M. Weismann

GSBS Dissertations and Theses

GM1 gangliosidosis is a lysosomal storage disorder caused by a deficiency in the catabolizing enzyme β-galactosidase (βgal). This leads to accumulation of GM1-ganglioside (GM1) in the lysosome inducing ER stress and cell death. GM1 gangliosidosis is primarily a disorder of the central nervous system (CNS) with peripheral organ involvement. In this work we report two major findings, 1) systemic treatment of GM1 gangliosidosis with an adenoassociated virus (AAV9) encoding mouse-βgal (mβgal) in a GM1 gangliosidosis mouse model (βGal-/-), and 2) an investigation into an intracranial injection of a therapeutic AAVrh8 encoding mβgal. Systemic treatment of GM1 gangliosidosis with AAV9 resulted ...


Hypertensive Acute Decompensated Heart Failure Presentations: On The Decline? : A Master's Thesis, Chad E. Darling Jul 2014

Hypertensive Acute Decompensated Heart Failure Presentations: On The Decline? : A Master's Thesis, Chad E. Darling

GSBS Dissertations and Theses

Background: The initial systolic blood pressure (SBP) in patients with acute heart failure (AHF) can be used as a guide when choosing specific pharmacologic treatments by helping identify the underlying type of HF (e.g., HF with preserved ejection fraction). Clinical experience and research data from our medical center suggests that AHF with elevated SBP may be presenting less frequently than in the past. This may call into question the utility of initial SBP as a clinical guide. The goal of this Master’s Thesis is to test the hypothesis that the frequency of AHF patients with a SBP>160mmhg ...


Clinical And Financial Impact Of Hospital Readmissions Following Colorectal Resection: Predictors, Outcomes, And Costs: A Thesis, Rachelle N. Damle Jun 2014

Clinical And Financial Impact Of Hospital Readmissions Following Colorectal Resection: Predictors, Outcomes, And Costs: A Thesis, Rachelle N. Damle

GSBS Dissertations and Theses

Background: Following passage of the Affordable Care Act in 2010, 30-day hospital readmissions have come under greater scrutiny. Excess readmissions for certain medical conditions and procedures now result in penalizations on all Medicare reimbursements. We examined the risk factors, outcomes, and costs of 30-day readmissions after colorectal surgery (CRS).

Methods: The University HealthSystem Consortium database was queried for adults (≥ 18 years) who underwent colorectal resection for cancer, diverticular disease, inflammatory bowel disease, or benign tumors between January 2008 and December 2011. Our outcomes of interest were readmission within 30-days of the patient’s index discharge, hospital readmission outcomes, and total ...


Adhd-200 Patient Characterization And Classification Using Resting State Networks: A Dissertation, Suzanne M. Czerniak Mar 2014

Adhd-200 Patient Characterization And Classification Using Resting State Networks: A Dissertation, Suzanne M. Czerniak

GSBS Dissertations and Theses

Attention Deficit/Hyperactivity Disorder (ADHD) is a common psychiatric disorder of childhood that is characterized by symptoms of inattention, impulsivity/hyperactivity, or a combination of both. Intrinsic brain dysfunction in ADHD can be examined through various methods including resting state functional Magnetic Resonance Imaging (rs-fMRI), which investigates patients’ functional brain connections in the absence of an explicit task. To date, studies of group differences in resting brain connectivity between patients with ADHD and typically developing controls (TDCs) have revealed reduced connectivity within the Default Mode Network (DMN), a resting state network implicated in introspection, mind-wandering, and day-dreaming. However, few studies ...


A Role For Neuronal Nicotinic Acetylcholine Receptors In Dopamine-Mediated Behaviors And The Hypnotic Response To Anesthetics: A Dissertation, Lindsey G. Soll Dec 2013

A Role For Neuronal Nicotinic Acetylcholine Receptors In Dopamine-Mediated Behaviors And The Hypnotic Response To Anesthetics: A Dissertation, Lindsey G. Soll

GSBS Dissertations and Theses

Neuronal nicotinic acetylcholine receptors (nAChRs) are ligand-gated cation channels that most notably influence dopamine (DA) release. In this thesis, I examine the role of nAChRs in mediating DA-related behaviors such as movement and drug dependence. To accomplish this, I utilized a “gain-offunction” knock-in mouse (the Leu9’Ala line) containing agonist-hypersensitive α4* nAChRs (* indicates other nAChR subunits in addition to α4 are within the receptor complex) that renders receptors 50-fold more sensitive to nicotine and acetylcholine than wild-type (WT) receptors. I found that DHβE, a selective antagonist for α4β2* nAChRs, induced reversible and robust motor dysfunction characterized by hypolocomotion, akinesia, catalepsy ...


Hepatitis C Virus Non-Structural Protein 3/4a: A Tale Of Two Domains: A Dissertation, Cihan Aydin Aug 2012

Hepatitis C Virus Non-Structural Protein 3/4a: A Tale Of Two Domains: A Dissertation, Cihan Aydin

GSBS Dissertations and Theses

Two decades after the discovery of the Hepatitis C Virus (HCV), Hepatitis C infection still persists to be a global health problem. With the recent approval of the first set of directly acting antivirals (DAAs), the rate of sustained viral response for HCV-infected patients increased significantly. However, a complete cure has not been found yet. Drug development efforts primarily target NS3/4A protease, bifunctional serine protease-RNA helicase of HCV. HCV NS3/4A is critical in viral function; protease domain processes the viral polyprotein and helicase domain aids replication of HCV genome by unwinding double stranded RNA transcripts produced by NS5B ...


Psychosocial And Behavioral Determinants Of Medication Nonadherence Among African Americans With Hypertension: A Dissertation, Yendelela L. Cuffee Aug 2012

Psychosocial And Behavioral Determinants Of Medication Nonadherence Among African Americans With Hypertension: A Dissertation, Yendelela L. Cuffee

GSBS Dissertations and Theses

The overarching goal of this dissertation was to elucidate the psychosocial and behavioral determinants of medication nonadherence among African Americans with hypertension. One in three Americans in the United States has hypertension, and the prevalence of hypertension among African Americans is among the highest in the world. In addition to healthy behaviors such as following a low-salt and low-fat diet, getting regular exercise, and reducing stress, patients with hypertension must also adhere to antihypertensive medications. Poor medication adherence may be driven by psychosocial and behavioral factors; however, the impact of these factors on medication adherence is unclear especially within the ...


The Role Of Itk In The Development Of Gamma Delta Nkt Cells: A Dissertation, Catherine C. Yin Aug 2012

The Role Of Itk In The Development Of Gamma Delta Nkt Cells: A Dissertation, Catherine C. Yin

GSBS Dissertations and Theses

The immune system is a complex network of interacting cells and tissues that is designed to protect the body from pathogens and other foreign substances. T cells are a major component of the immune system and consist of two distinct lineages distinguished by the expression of αβ or γδ T cell receptors (TCR). The Tec family kinase, Itk is an important mediator of signaling downstream of the TCR. Past studies on Itk has focused on how Itk regulates development, activation and differentiation of conventional αβ T cells and more recently how Itk regulates the development of innate-like αβ T cells ...


Studies On Cellular Host Factors Involved In The Hiv-1 Life Cycle: A Dissertation, Anna Kristina Serquiña Aug 2012

Studies On Cellular Host Factors Involved In The Hiv-1 Life Cycle: A Dissertation, Anna Kristina Serquiña

GSBS Dissertations and Theses

Human Immunodeficiency Virus Type 1 (HIV-1) is the causative agent of Acquired Immunodeficiency Syndrome (AIDS), currently the leading cause of death from infectious diseases. Since HIV-1 co-opts the host cellular machinery, the study of cellular factors involved is a rational approach in discovering novel therapeutic targets for AIDS drug development. In this thesis, we present studies on two such proteins. APOBEC3G is from the family of cytidine deaminases known to keep endogenous retroviruses and retrotransposons at bay to maintain stability of the human genome. APOBEC3G targets Vif-deficient HIV-1 particles and renders them noninfectious, partially through deaminase-dependent hypermutation of the provirus ...


Protein Ligand Interactions Probed By Nmr: A Dissertation, Jennifer M. Laine Jul 2012

Protein Ligand Interactions Probed By Nmr: A Dissertation, Jennifer M. Laine

GSBS Dissertations and Theses

Molecular recognition, defined as the specific interactions between two or more molecules, is at the center of many biological processes including catalysis, signal transduction, gene regulation and allostery. Allosteric regulation is the modification of function caused by an intermolecular interaction. Allosteric proteins modify their activity in response to a biological signal that is often transmitted through the interaction with a small effector molecule. Therefore, determination of the origins of intermolecular interactions involved in molecular recognition and allostery are essential for understanding biological processes. Classically, molecular recognition and allosteric regulation have been associated to structural changes of the system. NMR spectroscopic ...


Microrna Markers Of Acetaminophen Toxicity: A Master's Thesis, Jeanine Ward Jul 2012

Microrna Markers Of Acetaminophen Toxicity: A Master's Thesis, Jeanine Ward

GSBS Dissertations and Theses

Background
To investigate plasma microRNA (miRNA) profiles indicative of hepatotoxicity in the setting of lethal acetaminophen (APAP) toxicity in mice.

Methods
Using plasma from APAP poisoned mice, either lethally (500 mg/kg) or sublethally (150 mg/kg) dosed, we screened commercially available murine microRNA libraries (SABiosciences, Qiagen Sciences, MD) to evaluate for unique miRNA profiles between these two dosing parameters.

Results
We distinguished numerous, unique plasma miRNAs both up- and down-regulated in lethally compared to sublethally dosed mice. Of note, many of the greatest up- and down-regulated miRNAs, included, but were not limited to, 574-5p, 466g, 466f-3p, 375, 29c, and ...


Rna-Sensing Pattern Recognition Receptors And Their Effects On T-Cell Immune Responses: A Dissertation, Rachel F. Madera Jul 2012

Rna-Sensing Pattern Recognition Receptors And Their Effects On T-Cell Immune Responses: A Dissertation, Rachel F. Madera

GSBS Dissertations and Theses

Virus infection is sensed by the innate immune system through germline encoded pattern recognition receptors (PRRs). Toll-like receptors (TLRs), retinoic acid-inducible gene-I-like receptors (RLRs) and nucleotide-binding oligomerization domain-like receptors (NLRs) serve as PRRs that recognize different viral components. Microbial nucleic acids such as Ribonucleic acid (RNA) are important virus-derived pathogen-associated molecular patterns (PAMPs) to be recognized by PRRs. Virus recognition may occur at multiple stages of the viral life cycle. Replication intermediates such as single-stranded RNA (ssRNA) and double-stranded RNA (dsRNA) are detected by the RNA-sensing PRRs that initiate innate and adaptive immune responses. Triggering of the innate immune system ...


Therapeutic Silencing Of Mutant Huntingtin By Targeting Single Nucleotide Polymorphisms: A Dissertation, Edith L. Pfister Jul 2012

Therapeutic Silencing Of Mutant Huntingtin By Targeting Single Nucleotide Polymorphisms: A Dissertation, Edith L. Pfister

GSBS Dissertations and Theses

Huntington’s disease (HD) is an autosomal dominant, progressive neurodegenerative disorder. Invariably fatal, HD is caused by expansion of the CAG repeat region in exon 1 of the Huntingtin gene which creates a toxic protein with an extended polyglutamine tract 1. Silencing mutant Huntingtin messenger RNA (mRNA) is a promising therapeutic approach 2-6. The ideal silencing strategy would reduce mutant Huntingtin while leaving the wild-type mRNA intact. Unfortunately, targeting the disease causing CAG repeat expansion is difficult and risks targeting other CAG repeat containing genes.

We examined an alternative strategy, targeting single nucleotide polymorphisms (SNPs) in the Huntingtin mRNA. The ...


Slow-Cycling Cancer Cells: A Dissertation, Nathan F. Moore Jun 2012

Slow-Cycling Cancer Cells: A Dissertation, Nathan F. Moore

GSBS Dissertations and Theses

Tumor recurrence after chemotherapy is a major cause of patient morbidity and mortality. Recurrences are thought to be due to small subsets of stem-like cancer cells that are able to survive chemotherapy and drive tumor re-growth. A more complete understanding of stem-like cancer cell regulation is required to develop therapies to better target and eliminate these cells.

Slow-cycling stem cells are integral components of adult epithelial tissues and may give rise to cancer stem cell populations that share similar characteristics. These slow-cycling adult stem cells are inherently resistant to traditional forms of chemotherapy and transference of this characteristic may help ...


Characterization Of Drug Resistance In Mycobacterium Tuberculosis Via Saturating Mutagenesis Of Drug Targets: A Master’S Thesis, Michelle J. Harris Jun 2012

Characterization Of Drug Resistance In Mycobacterium Tuberculosis Via Saturating Mutagenesis Of Drug Targets: A Master’S Thesis, Michelle J. Harris

GSBS Dissertations and Theses

Mycobacterium tuberculosis isolates from multiple drug resistant or extensively drug resistant patients show a particular set of mutations in drug targets conferring resistance. However, the selection of drug-resistant strains in vitro yields an alternative set of mutations, thought to result from the cost-benefit associated with drug resistance. Mutations allowing for survival under antibiotic may not be beneficial when presented with the host environment or with a drug-free environment. These fitness effects drive the natural evolution of this bacterium. Using recombineering a large cohort of mutations was generated within two drug targets, inhA and gyrA, to study in vitro the variability ...


Small Molecule Investigation Of Kcnq Potassium Channels: A Dissertation, Karen Mruk May 2012

Small Molecule Investigation Of Kcnq Potassium Channels: A Dissertation, Karen Mruk

GSBS Dissertations and Theses

Voltage-gated K+ channels associate with multiple regulatory proteins to form complexes with diverse gating properties and pharmacological sensitivities. Small molecules which activate or inhibit channel function are valuable tools for dissecting the assembly and function of these macromolecular complexes. My thesis focuses on the discovery and use of small molecules to probe the structure and function of the KCNQ family of voltage-gated K+ channels.

One protein that obligatorily assembles with KCNQ channels to mediate proper assembly, trafficking, and gating is the calcium sensor, calmodulin. Although resolution of the crystal structures of calmodulin associated with isolated peptide fragments from other ion ...