Randi J. Hagerman Summer Scholars Research Awards

Watch each of the 2024 Summer Scholar 15-minute presentations.

Emily Peery

Women’s Healthcare Provider Knowledge on FXPOI: Improving Understanding Through an Educational Tool

Affiliation: First-year graduate student, Emory University, Genetic Counseling Training Program

Supervisor: Emily Allen, PhD

Project Summary: Individuals with Fragile X-associated primary ovarian insufficiency have reported needing to visit multiple providers and advocate for their own diagnosis and care because providers were unaware of Fragile X-associated disorders. FXPOI is defined as irregular (i.e., cycling every 4-6 months) or cessation of menses before 40 (premature menopause) in females who carry an FMR1 premutation. Prior research showed the average amount of time from symptom onset to FXPOI diagnosis was about one year. Due to this provider gap in knowledge, further research was done to identify what these gaps in knowledge specifically are amongst women’s healthcare providers.

The study goal is to target these knowledge gaps through questions in a pre-test survey and assess if women’s healthcare provider knowledge of FXPOI is improved after reviewing the educational tool. A post-test survey will be sent to the providers one month after reviewing the tool. We will also look at whether demographics affect FXPOI knowledge and what general carrier screening practices and provider workups for primary ovarian insufficiency are. The long-term goal would be for women’s healthcare providers to continue to refer to and share this educational tool to shorten the time to diagnosis for women with FXPOI and contribute to an improved quality of life and reproductive outcomes.

Manasi Inamdar

Identifying Translational Dysregulation Underlying Auditory System Dysfunction in a Rat Model of Fragile X Syndrome

Affiliation: Second-year graduate student, University of Illinois Urbana-Champaign, Molecular and Cellular Biology

Supervisor: Benjamin D. Auerbach, PhD

Project Summary: Fragile X syndrome is the most common inherited cause of autism, with a mutation in the FMR1 gene that encodes for a critical RNA-binding protein FMRP. Genes in our body contain instructions from DNA to make proteins, which are essential for building cells in our bodies. These cells, particularly neurons, form connections called synapses that allow them to communicate and create networks. Within these networks, proteins play a critical role in maintaining and shaping circuits in the brain. Neural circuits are a group of interconnected neurons that work together like a complex web of pathways to process information and give rise to behavior.

For developing effective therapies for treating FXS, it is crucial to study how disruptions across levels of neuronal function (molecular, cellular, circuit) ultimately lead to abnormal processing of sensory information and social, communication alterations seen in FXS patients as well as FXS rodent models. Atypical sensory processing is a core feature of FXS and markedly elevated sensitivity to sound stimuli is the most prevalent and debilitating symptom of FXS. By studying how sound processing is altered in FXS rat models, we seek to determine if these changes stem from disruptions in protein synthesis, a crucial process for proper neuronal function.

Increased synthesis of proteins is pathological in FXS, and by understanding what these proteins are and why they are problematic, we can start to understand the factors impacting FXS such as underlying neural circuits. This summer project proposal will determine if auditory processing deficits in FXS are the result of cellular-molecular changes in the auditory system. Ultimately, our goal is to contribute to the development of therapies that can improve the lives of individuals and families affected by FXS.

Emily Timm

The Neural Mechanisms Underlying Cortical Control of Gait and Cognition in Fragile X-Associated Tremor/Ataxia Syndrome (FXTAS)

Affiliation: Third-year PhD student, Rush University Medical College, Integrated Biomedical Sciences Program

Supervisor: Joan O’Keefe, PT, PhD

Project Summary: Little is known about the changes in the brain that cause the major symptoms of FXTAS, including problems with balance, walking, and memory. By using a non-invasive brain imaging cap, our team can measure activity in special areas of the brain that play roles in memory, attention, and the planning of movements. We ask participants to wear this brain imaging cap while performing activities that are affected by FXTAS, such as walking under simple and more difficult conditions and completing a series of challenging cognitive tests. All study participants also have an MRI scan to take pictures of the brain, so that we can determine any connections between brain activation and structure.

This research study is important because our findings will provide more information about how the disease develops and can assist with the development of effective treatments for FXTAS, which currently do not exist.

Thomas Christensen

Exploring Social Determinants of Health and the FMR1 Premutation Symptomology in Women

Affiliation: Second-year graduate student, University of South Carolina, Communication Sciences and Disorders

Supervisor: Jessica Klusek, PhD

Project Summary: We already know that where someone lives can have a big impact on their health. Because women with the FMR1 premutation already have an increased risk of a variety of health issues due to genetic factors, it is important to also understand how one’s neighborhood can impact those health issues, for better or for worse. This would help researchers and advocacy organizations know where to allocate their limited resources to have the greatest benefit on the health of women with the FMR1 premutation.

Watch each of the 2023 Summer Scholar 15-minute presentations.

Aditi Mahajan

The Electroretinogram and FMRP: Correlating Biomarkers for Fragile X Disorders

Affiliation: Senior at the University of California, Davis, majoring in Global Disease Biology and minoring in History

Supervisor: Dr. Randi J. Hagerman, University of California, Davis, Developmental and Behavioral Pediatrics

Project Summary: The Electroretinogram and FMRP project will assess whether electroretinograms, which measure the activity of the retina, can be used as a reliable marker to detect deficits in FMRP in both those with FXS and those with premutation involvement. The b-wave deficits in the ERG may also correlate with other behavior problems and help us understand how BK channel deficits relate to the phenotype of FXS and premutation involvement. The study will involve a series of questionnaires and assessments, a clinical ERG reading, and molecular analyses. This study can potentially lead to an easy and quick way to screen for Fragile X involvement or FMRP deficits and how this affects behavior.

In Their Own Words: “I worked with Dr. Randi Hagerman at the UC Davis MIND Institute during my final year as an undergraduate at UC Davis. During my fellowship I had the unique opportunity to interact directly with the Fragile X community through my work as a clinical researcher. I obtained electroretinogram readings, learned how to quantify FMRP levels in blood samples, administered sensory and adaptive questionnaires, and recognized the significance of BK channel function in those with Fragile X.

“I have come to learn the Fragile X community is based on a strong foundation of teamwork and support. The mentorship I received through the summer fellowship allowed me to develop my professional skills in research and appreciate the importance of interdisciplinary collaboration. The cutting-edge research at the MIND Institute represents hope and possibility for the Fragile X community.

“I am so grateful to have had the chance to work with Dr. Randi Hagerman and the NFXF towards the shared goal of improving the lives of those affected by Fragile X.”

Alexandra Singleton

FXPOI Survey in Women’s Healthcare Providers

Affiliation: First-year student in the Emory University Genetic Counseling Training Program

Supervisor: Emily G. Allen, PhD, Emory University, Department of Human Genetics

Project Summary: Patients with Fragile X-associated primary ovarian insufficiency (FXPOI) have been reported to have a long period of time between the onset of symptoms and receiving a diagnosis, with an average time of over a year. FXPOI is defined as menopause before age 40 caused by a premutation in the FMR1 gene.

Several patients have reported having to bring educational materials to their providers because they were unaware of Fragile X-associated disorders.

The goals of this study are to identify the current knowledge base of FXPOI in women’s healthcare providers and to see if provider demographics affect their knowledge of FXPOI as well as to identify POI related genetic testing and carrier screening patterns in their practice.

In Their Own Words: “Our study aims to identify where gaps in knowledge of Fragile X-associated primary ovarian insufficiency (FXPOI) lie amongst women’s healthcare providers including physicians, advanced practice nurses, and medical students. Past studies have found that the average time to a FXPOI diagnosis after symptoms begin is over one year. Additionally, patients have reported bringing educational materials to their providers because they were unaware of Fragile X-associated disorders.

“This summer, we distributed a survey via email to women’s healthcare providers and at a conference targeted toward OBGYN providers that assessed FXPOI knowledge, familiarity with primary ovarian insufficiency, reproductive carrier screening practices, and genetics exposure.

“By identifying these gaps in provider knowledge, our goal is to shorten the time to FXPOI diagnosis and improve quality of care for FXPOI patients and their families through provider-focused educational materials that will be created as part of a future project.”

Maureen Butler

The Biological Basis of Pragmatic Language in Fragile X Premutation Carriers

Affiliation: First-year doctoral student in the Clinical Psychology program at the Northwestern University Feinberg School of Medicine

Supervisor: Dr. Molly Losh, Northwestern University, Communication Sciences and Disorders

Project Summary: Pragmatic language, or how we use language in social contexts like conversations, is a complex language skill that allows us to build relationships and connect with others. Individuals with fragile X syndrome have difficulties with pragmatic language and often, people with the fragile X premutation (PM) have differences in how they use pragmatic language.

Despite this, little is understood about the biological and genetic foundations of pragmatic language skills in women with the PM. This project aims to understand how biological factors might influence pragmatic language skills in PM carriers. To do this, we will examine the relationship between FMR1 genetic variation, a measure that indicates how the brain hears and represents sound, and pragmatic language abilities.

Through this research, we hope to gain insight into the causes of language differences in FMR1 conditions. By understanding the root of these differences, we may be able to develop target language interventions, as well as investigate the relationship between pragmatics and clinical disorders experienced by many PM carriers such as anxiety and executive function difficulties, ultimately creating a path towards targeted treatment.

In Their Own Words: “Participating in the National Fragile X Foundation Randi J. Hagerman Summer Scholars Award has been a truly incredible experience. The program has provided me the opportunity to conduct exciting, innovative research investigating the biological bases of social communication abilities in women who carry the FMR1 premutation.

“Through this work, I have started examining the biological foundations of prosodic synchrony, an important skill involved in social communication, through the novel application of a statistical analysis that incorporates the temporal dynamics of speech. The preliminary results discussed in my presentation show that this methodology may be sensitive in detecting subtle language differences in premutation carriers.

“This is a vital first step in developing social communication interventions for individuals with FMR1 conditions that the target underlying causes of language difficulties. As I continue to advance this project, I hope to publish my results in an academic journal. Not only has the Randi J. Hagerman Summer Scholars program given me with the opportunity to initiate this exciting project, but it has also provided an important platform to share this research with the Fragile X community.

“For me, engaging with and sharing my work with the community has been the most rewarding and impactful element of this program. I am so grateful for this experience and am honored to have been selected to participate in the Randi J. Hagerman Summer Scholars program.”

Watch each of the 2022 Summer Scholar 15-minute presentations.

Andy King

The Personal Healthcare Experiences of African American Women Who Carry a Fragile X Premutation

Affiliation: Emory University Genetic Counseling Training Program

Supervisor: Dr. Emily G. Allen

Project Summary: Our study explores the personal healthcare experiences of African American women who carry a Fragile X premutation. We conducted phone interviews with 20 African American women with a premutation. We asked about the barriers to accessing and receiving adequate healthcare that they have experienced, including treatment of any Fragile X-associated primary ovarian insufficiency (FXPOI) symptoms they may have.

Our study highlights particular healthcare disparities experienced by underrepresented women in Fragile X research and improves health outcomes for all women with a premutation.

In their own words: “Our study investigates the personal healthcare experiences of African American women who carry the Fragile X premutation. Throughout the summer and continuing this fall, we are interviewing 8–10 women with a premutation to understand their experiences with healthcare, any barriers they have had in their care, their emotional health, and other factors impacting their experiences.

“Our preliminary results show that African American women experience a worry over their symptoms being taken seriously. This is a similar experience to other women studied previously. They have expressed the difficulties in sharing their diagnosis with family, which has led to lack of support and strained relationships. We have also found that most participants endorse experiencing anxiety and depression throughout their experiences, with various approaches to care that has helped address these symptoms.

“We hope our research deepens our knowledge of underrepresented women’s experiences in genetic research, particularly in the Fragile X community.”

Natalia Rivera Alfaro

Searching FXTAS Genetic Modifiers by Combining Whole Genome Sequencing and Drosophila Model

Affiliation: Universidad Nacional Autónoma de México/Emory University School of Medicine, Department of Human Genetics

Supervisor: Dr. Peng Jin

Project Summary: Fragile X-associated tremor/ataxia syndrome (FXTAS) is a neurodegenerative genetic disorder. People with FXTAS suffer from cognitive decline, tremors and ataxias (difficulty with balance and coordination). It is caused by CGG trinucleotide repeats, which are associated with the Fragile X mental retardation gene (FMR1).

The number of CGG repeats can cause Fragile X syndrome (with over 200 repeats) or FXTAS (between 55 and 199 repeats). However, people with the same number of repeats can show differences in the degree of expression of certain traits, this means that one person can have complete FXTAS and another one only mild symptoms even though they have the same number of repeats (this is known as incomplete penetrance).

We hypothesize that there may be other genes involved in FXTAS that affect how much it is expressed. We use the genomic information of 100 patients with FXTAS and 100 controls with mild expression of FXTAS. We then compare patients and controls to find a list of candidate genes that may be modulating the incomplete penetrance. Finally, we use fly models to see what happens if each candidate gene is overexpressed or depleted in combination with FXTAS. This allows us to determine which of the candidate genes are involved in the modulation of FXTAS.

This study will help interpret the genetic architecture of neurodegeneration, both associated with FXTAS and ataxia in general, as well as provide the opportunity for early intervention that may delay or prevent the onset of FXTAS.

In their own words: “This summer I had the amazing opportunity of working with fly models in order to investigate the mechanisms responsible for the incomplete penetrance seen in FXTAS patients. I worked with various fly lines that knock down genes that we know are different between patients with mild FXTAS symptoms and patients with more severe symptoms in order to test their interaction with the CGG repeats. This will hopefully give us some interesting genes to look at more in depth in order to understand the reasons behind the incomplete penetrance of FXTAS.

“As an international student I also learned a lot from the American culture and work style and I am overall very thankful for this opportunity.”

Dominic DeBiasi

Unraveling the Role of Repetitive RNA and RANprotein in FXTAS

Affiliation: University of Michigan

Supervisor: Dr. Peter Todd

Project Summary: Fragile X-associated tremor/ataxia syndrome (FXTAS) is a neurodegenerative disease that stems from multiple copies of a short sequence in the FMR1 gene. Within a mid-range of copies, there exists an unorthodox way to make protein that would not be expected to be produced otherwise.

Whether this “new” protein plays a role in the disease pathology remains unknown; therefore, this project provides insight as to whether this protein is sufficient to drive neurodegenerative-like phenotypes.

In their own words: “I am incredibly grateful to have been selected as a Randi J. Hagerman National Fragile X Foundation Summer Scholar. The award provided me with the opportunity to engage in exciting research and gain experience working in the lab, but more importantly, introduced me to the wonderful Fragile X community.

“Throughout the course of this project, I’ve learned that community outreach and communication is just as important as the scientific research itself, if not more so. Organizations like the NFXF help to bring issues of human disease and medicine to the forefront, creating a bridge between the scientific community and the public, which results in huge impacts for treating these diseases that afflict so many.”

Jordan Norris

ROC Analysis of Biomarker Combination in Fragile X Syndrome-Specific Clinical Trials: Evaluating Treatment Efficacy via Exploratory Biomarkers

Affiliation: University of Oklahoma

Supervisor: Dr. Lauren Ethridge

Project Summary: Clinical trials, specifically phase 2 clinical trials, are important steps in the process of developing novel treatments that are specific to Fragile X syndrome (FXS) and treating underlying causes of FXS over treating symptoms, alone. The goal of treating the source of FXS is to create long-term shifts that improve the quality of life of patients with FXS and their loved ones. Phase 2 clinical trials aim to establish safety and tolerability, but also give an indication in a relatively small group of individuals of whether the treatment is working.

A recent phase 2 clinical trial of the drug BPN14770 showed cognitive improvements through treatment, but measures of drug effects on brain function were relatively small. The study aims to revisit these physiological measures and assess them using a classification method that combines multiple brain function measures. We propose that combinations of brain function measures may give us a better idea of the complex interplay between individual characteristics of those with FXS and their response to treatment.

In their own words: “Being an NFXF Randi Hagerman Summer Scholar recipient is an amazing opportunity to work on a project that may have important implications for Fragile X Syndrome–specific clinical trial methods and for the specific outcomes of the recent, successful phase 2 clinical trial testing BPN14770 (now zatolmilast). Specifically, it gave me the time to take a data-driven approach to biomarker selection where multiple biomarkers can be assessed without additional statistical strain.

“Clinical trials, specifically phase 2 clinical trials, are important steps in the process of developing novel treatments that are specific to FXS and treating underlying causes of FXS over treating symptoms, alone. Developing feasible methods for FXS-specific clinical trials is an important step in evaluating the gap between known pharmaceutical mechanisms and measurable behaviors/clinical outcomes, especially when methodological issues arise.

“The chance to serve as a summer scholar allowed me to take these important first steps with existing clinical trial data with the hope that these results make a lasting impact on the way that we approach the measurement of pharmaceutical effects in FXS clinical trials.”

Jessica Tang

Transcriptional Gene Silencing of FMR1 as a Treatment for Fragile X-Associated Tremor/Ataxia Syndrome

Affiliation: University of California Davis School of Medicine

Supervisor: Dr.Paul Hagerman

Project Summary: Fragile X-associated tremor/ataxia syndrome (FXTAS) is caused by an abnormal Fragile X (FMR1) gene that produces high levels of the Fragile X protein (FMRP) that leads to neurodegeneration. We propose a gene therapy to knock out the abnormal FMR1 gene and, in follow-on work, add a coding cassette comprising a mini-promoter and FMRP coding region.

The goal of my project is to identify the RNA sequence(s) that can successfully target and silence the abnormal FMR1 gene. To do this, I package multiple RNA candidates into small (nano)particles that are then delivered to fibroblast cells. After the fibroblasts have grown for 1 week, 4 weeks, and 8 weeks, I will quantify how much FMR1 RNA and protein are produced by the fibroblasts. This allows us to determine how effective a given candidate sequence is in silencing the FMR1 gene.

From my results, we will select the candidate RNA to be used in subsequent studies to silence FMR1 in rat and rhesus models. Our long-term goal is to develop a genetic therapy that will prevent neurodegenerative difficulties in patients with FXTAS by targeting the expression of the abnormal FMR1 gene.

In their own words: “During my first year of medical school, I worked with Dr. Randi Hagerman, helping out with clinical trials and shadowing her through patient visits. This summer through the RJH NFXF Summer Scholar Program, I had the amazing opportunity to work under Dr. Paul Hagerman and see the translational science side of research — something that is often difficult to do during the school year.

“This summer, I am proud to have created a pipeline for identifying and screening small transcriptional gene silencing-RNAs (sTGSs) for gene therapy in Fragile X and FXTAS. Furthermore, I was able to identify two potential sTGSs that could lead to long-term suppression of abnormal FMR1 gene.

“Thank you NFXF for providing a platform to share my work and for supporting my aspirations of becoming a physician in academic medicine. I am so excited to continue working in the field of Fragile X among amazing mentors and leaders of their field!”

Collis Brown

Potential Drug Therapy for Fragile X Tremor/Ataxia Syndrome

Affiliation: Howard University, Department of Pharmacology

Supervisor: Dr. Tamaro Hudson

Project Summary: As a recipient of the National Fragile X Summer Scholars Award, this summer has been nothing short of amazing. My project is titled “Potential Drug Therapy for Fragile X Tremor/Ataxia Syndrome.” There are currently no effective treatments for progression of the disease or cures.

Mitochondrial damage is a recently identified cellular feature of both FXTAS-diagnosed and asymptomatic expansion carriers. FXTAS-derived fibroblasts and neurons have reduced levels of key mitochondrial proteins, increased production of reactive oxygen species, and decreased ATP production. Therefore, the reversal of mitochondrial dysfunction may serve as an early pharmacological intervention in the treatment of FXTAS.

My summer research entailed analyzing the biological efficacy of arachidonyldopamine, macamide, and their analogues in normal human fibroblast and neurons after glucose oxidase induced oxidative damage.

In order to establish proof of principle to induce mitochondrial damage, I focused on determining at what concentration does glucose induce cytotoxicity or disruption in cell viability. I also assessed if this concentration was time dependent.  Then, I wanted to determine whether the natural compounds MSKE and Curcumin effect cell viability in human dermal fibroblast cells at variant concentrations over time. Lastly, I examined if the natural compounds MSKE and Curcumin can inhibit the glucose induce disruption of cell viability.

From the results, the current concentration of glucose proved to be ineffective in inducing physiological disruption of cell viability. However, we found an increase in cell viability with the use of Curcumin at 12.5 and 50nM. Curcumin in combination with glucose also showed a similar trend.

Also, the introduction to a network of colleagues this summer was phenomenal, even though each of us are studying different areas of Fragile X all of our goals remain the same, which is to make an impact within the Fragile X community.

John Burwinkle

MicroRNA-Mediated Correction of Dendritic Spine Abnormalities in a Mouse Model for FXS via Modulation of Cytoskeletal Proteins

Affiliation: Cincinnati Children’s Hospital Medical Center

Supervisor: Dr. Christina Gross

Project Summary: Through the NFXF’s Summer Scholar Research Award I was fortunate enough to get a deeper understanding of how research in the basic sciences can translate to future practices in clinical medicine by revealing mechanisms that could be targets for potential therapeutics.

I researched how inhibiting a microRNA, miR-324-5p, via an antagomir effects neuronal structure and protein expression in a Fragile X mouse model. The antagomir treatment in FMR1 knockout mice did decrease dendritic spine density that is typically higher in these mice than wild-type controls. This provides initial preclinical evidence for the use of antagomirs as a potential therapeutic for Fragile X syndrome. However, the mechanism and targets of miR-324-5p remain unclear and should be further studied before the therapeutic should be used.

This in combination with hearing the stories of the families. Our networking event made me appreciate how my work fits into the larger Fragile X community and provides hope for the future. Ultimately, this summer displayed how intertwined research is and how everyone from both the basic sciences and clinical realms play a significant role in the future of medicine.

Bonnie McKinnon

The Diagnostic Experience of Women with Fragile X-Associated Primary Ovarian Insufficiency (FXPOI)

Affiliation: Emory University, Department of Human Genetics

Supervisor: Dr. Emily Allen

Project Summary: Our study is exploring the diagnostic odyssey of women with Fragile X-associated primary ovarian insufficiency. This summer we interviewed 24 women with FXPOI to better understand their diagnosis, healthcare experience, self-advocacy, along with other factors that contribute to their experiences.

A lack of educational materials and clinician training might be contributing to the difficulty women experience receiving a FXPOI diagnosis and have a large psychosocial impact for women with FXPOI. It is essential that any healthcare provider ordering genetic testing for at-risk families understands the risks that come with FXPOI as well as how the healthcare system is impacting these families.

We hope this study will serve as a steppingstone to create a centralized and improved diagnostic experience for FXPOI.

Elizabeth Saoud

A Study of Visual Evoked Potentials in Children with Fragile X Syndrome

Affiliation: Boston Children’s Hospital, Harvard Medical School

Supervisor: Dr. Carol Wilkinson

Project Summary: This summer I conducted a study on visual evoked potentials (VEPs) in children with Fragile X syndrome. My project combined EEG methods and behavioral measures to identify differences in the VEP response that could serve as neural markers to be used in clinical trials and to monitor developmental progress in children with FXS.

In the coming months, I will be continuing data analysis for the project and examining additional behavioral measures to further characterize the relationship between VEP, cognitive development, and FXS.

I am thankful for the opportunity I had this summer, and I look forward to seeing the positive impact of this study in the future.

Andrew Snider

Investigation of the Potential of AZD7325 Treatment on EEG Oscillations, Gamma Waveforms, and Dendritic Spine Morphology in the Mouse Model of FXS

Supervisor: Christina Gross, PhD

Project Summary: We hypothesize that treatment with the GABAAα2/3 agonist AZD7325 will rescue increased gamma oscillations and abnormal dendritic spine morphology in FMR1 KO mice.

Aim 1 will analyze the effect of AZD7325 treatment on basal gamma EEG power in FMR1 KO mice and their WT littermates.

Aim 1 Approach: We will test the potential of the GABAAα2/3 agonist, AZD7325 (AstraZeneca), on normalizing basal EEG levels and, in particular, the increased gamma EEG power observed in FMR1 KO mice. 6-8-week-old FMR1 KO mice and their WT littermates will be implanted with cortical electrodes for continuous video/EEG recording using a wireless system from Data Sciences International. Three days after surgery, baseline EEGs will be recorded for a period of 7 days to confirm basal EEG waveforms in the mice. Then, the mice will be administered with daily injections 1mg/kg of AZD7325 or the vehicle control (0.05% SBECD (Sulfobutyletherbetacyclodextrin)) in Milli-Q water for 10 consecutive days (dose established by previous studies in mice).

The recorded EEG data will be analyzed for individual waveforms, total power, and relative and absolute gamma oscillations using NeuroScore software. Feasibility of these experiments is confirmed by our previous publications using mouse EEG analyses (Gross et al., 2016), and preliminary data that recapitulate published data.

Aim 2 will examine the effect of treatment of AZD7325 on dendritic spine density and morphology.

Aim 2 Approach: Post completion of ten days of drug treatment and recording, the mice brains will be collected and preserved for analysis of dendritic morphology. The brains will be first treated through the Golgi staining process using the FD Rapid Golgi Staining Kit as we have done previously Gross et al., 2015b. Briefly brains will be sectioned using a vibratome (160 μm slices), stained according to the manufacturer’s protocol and dendritic spines of CA1 pyramidal neurons will be imaged using a widefield microscope using a 60X objective, and quantified using Image J (NIH).

The dendritic spine counting techniques are routinely performed in the laboratory (Gross et al., 2015b). Most recently, we have used a similar treatment paradigm (10 days of daily dosing) with a different drug to show that dendritic spine density was reduced after treatment. [Figures not included.]

Kara Brown

Brain Microstates as a Window into Sensory Sensitivity in Fragile X Syndrome

Supervisor: Dr. Lauren Ethridge

Project Summary: This study will be the first to observe the microstates in FXS in both the resting and prestimulus states. We aim to 1) assess whether FXS, similar to ASD, maintain more stable microstates during resting EEG, and whether this is correlated with behavioral or sensory rigidity; and 2) assess whether pre-stimulus microstates affect sensory processing in FXS specifically associated with sensory hypersensitivity.

We hypothesize that people with FXS will have more stable resting microstates and also will spend more time in an abnormal pre-stimulus microstate, potentially associated with increased activity in primary sensory cortices, leading to hyperexcitability and hypersensitivity to auditory stimuli.

Ryan Risgaard

Development and Screening of Novel Compounds for FMR1 Gene Reactivation

Affiliation: University of Wisconsin-Madison

Supervisor: Dr. Xinyu Zhao, Professor of Neuroscience

Project Summary: Developments in stem cell research and sequence-specific synthetic molecules have presented promising opportunities for potential drug therapies of Fragile X Syndrome (FXS). FXS is an X-linked genetic disease and the largest known cause of inherited intellectual disability.

Caused by a CGG repeat expansion in the FMR1 gene, FXS results in hypermethylation and subsequent shutdown of gene activity and protein expression. Past therapeutic strategies have attempted to restore FMR1 activity through drug compound screens, but to date, no compounds have successfully, fully reactivated the FMR1 gene.

To address this challenge, my mentor Dr. Zhao’s lab has created a luciferase-based reporter cell line that has allowed for the large-scale screening of compounds for FMR1 gene reactivation. Additionally, my co-mentor Dr. Ansari’s lab has successfully developed sequence-specific polyamides that have been shown to bind methylated DNA and reduce methylation of targeted sites.

Design of a novel polyamide compound that binds and reactivates the FMR1 gene would represent a significant step towards therapeutic strategies of FXS. Therefore, this project will focus on (1) conducting a secondary screening of potential small molecule gene reactivators and (2) aiding in the development of novel polyamide compounds that bind CGG repeats and reduce methylation of the FMR1 gene.

Ellie Eckert

Social Gaze Differences in Fragile X Syndrome and Idiopathic Autism Spectrum Disorder

Supervisor: Craig Erickson, MD

Project Summary: There are a number of noted similar behaviors and social impairments between individuals with Fragile X syndrome and Autism spectrum disorder, so much so that roughly 30% of children with FXS also receive a diagnosis of autism. This study will investigate the fundamental differences in mechanisms that drive these behavioral features seen in both FXS and ASD, and may provide the basis to develop tools that will accurately measure social deficits specific to FXS. These findings will allow for future investigation into distinguishing features between FXS and ASD that may aid in accurate diagnosis and tailored treatment.

Ndeye Marieme Ndiaye

Social Reward Learning in Human Patients with Autism

Supervisor: Gül Dölen, MD, PhD

Project Summary: The recent development of a human version of the conditioned place preference (CPP) assay we use to measure the rewarding properties of social interactions in mice, dramatically improves the translational validity of neural mechanisms and therapeutic targets uncovered using this assay. Preliminary data collected in the Dölen lab indicates that FMR1 knockout autism and Fragile X model mice have impaired social reward learning. In collaboration with the Thompson lab, I will conduct experiments to test the hypothesis that patients with Fragile X and autism have a similar impairment in social reward learning, using the human version of the social CPP assay.

Marwa Zafarullah

Genomic Editing of FMR1 Premutation in Human Fibroblast Cell Lines via Delivery of Purified Cas9 Ribonucleoproteins

Supervisor: Flora Tassone, PhD

Project Summary: This study will help us to better understand the impact of the CGG repeats on the different clinical premutation phenotypes and the molecular mechanisms behind the development of Fragile X-associated disorders. The CRISPR Cas9 technology has a profound impact on research efforts including identification of genes as well as the development of new disease models.

The genome editing by transient expression of Cas9 (as RNP) will permit the consideration of a range of delivery options for therapeutic applications. In the future, with the optimization of suitable delivery system and proper assessment of specificity, we can clinically translate this study for the therapeutic editing in the Fragile X-associated disorders.

Joseph Krzeski

Testing MicroRNA-Mediated Silencing of Kv4.2 as Therapeutic Target in an FXS Mouse Model

Supervisor: Dr. Christina Gross

Mika Nagamoto and Jenna Nagamoto

Evaluation of a Direct Outcome Measure of Social Interaction in Children and Adults with Fragile X Syndrome

Supervisor: Nicole Tartaglia, MD

Chandler Robinson

Activity-Dependent Dysfunction of Microglia in Fragile X Syndrome

Supervisor: Craig A. Erickson, MD

Seokyu (Philip) Shin

Understanding the Role of Agranular Insular Cortex Projections to the Nucleus Accumbens in the Pathogenesis of Blunted Social Affect in Fragile X

Supervisor: Dr. Gül Dölen

Kanisha Desai

Cellular and Biomolecular Characterization of Fragile X Patient Induced Pluripotent Stem Cell Derived Neurons

Affiliation: Emory University

Supervisor: Dr. Gary Bassell

Project Summary: The loss of the Fragile X mental retardation protein (FMRP) in Fragile X syndrome has been linked to an increase in protein synthesis and increased activity of the phosphoinositide-3 kinase (PI3K) signaling pathway. Previous work in the Bassell lab has shown that FMRP directly regulates the catalytic subunit of PI3K and that reduction of overactive PI3K signaling corrects specific deficits associated with FXS. During my summer project, I used control and FXS patient-derived induced pluripotent stem cells (iPSCs) to generate neurons. Work in progress has been to characterize cellular and morphological deficits in FXS iPSC-derived neurons and assess the potential therapeutic value of PI3K inhibition.

Nada El-Sayed

The Effect of Genetic Background on Kv4.2 Expression in FXS Mouse Models

Affiliation: Cincinnati Children’s Medical Hospital Center

Supervisor: Christina Gross, PhD

Project Summary: Fragile X syndrome (FXS) is often characterized by hyperactivity and is associated with epilepsy, especially in children. The underlying molecular causes of this hyperactivity remain unknown. Kv4.2, a protein that helps transmit signals between nerve cells, is a key regulator of nerve cell activity in the brain. My lab has previously shown that in a mouse model of FXS, levels of Kv4.2 are decreased. In pilot studies, which were part of my summer research, we showed similarly decreased Kv4.2 levels in another mouse model of FXS suggesting that reduced Kv4.2 function could contribute to neuronal hyperactivity and epilepsy in FXS.

My research this summer focused on regulation of Kv4.2 production, as identifying ways to increase Kv4.2 levels could potentially lead to the development of novel treatment strategies for FXS. I worked with microRNAs, which are natural and selective inhibitors of protein production that lead to reduced protein levels of their targets. I was able to confirm regulation of Kv4.2 by a specific microRNA. By inhibiting this microRNA in the future, we could potentially increase Kv4.2 expression in patients with FXS, which may help to reduce neuronal hyperactivity.

Sarah Fitzpatrick

Institute A Controlled Trial of Sertraline in Young Children with Fragile X Syndrome

Affiliation: UC Davis MIND Institute

Supervisor: Randi Hagerman, MD

Project Summary: My project involved managing the extensive process of completing a trial of sertraline in young children with FXS by bringing in the final subjects and preparing the data for analysis. The analysis is complete and we are currently writing the paper for publication. In addition, I collaborated with many brilliant people, shadowed Dr. Hagerman in patient visits, learned about the Fragile X field, and developed important research, clinical, and management skills. Ultimately, I discovered my passion for working with families affected by Fragile X and greatly look forward to specializing in FXS as a physician and researcher in the future.

Erin Robertson-Dick

Effects of Cognitive Function and Dual-Task Interference on Balance and Gait in Premutation Carriers of the Fragile X Mental Retardation 1 Gene

Affiliation: Rush University Medical Center

Supervisors: Joan A. O’Keefe, PT, PhD and Deborah A. Hall, MD, PhD

Project Summary: My summer project was to determine the impact of cognitive interference on balance and gait function in carriers of a premutation on the Fragile X mental retardation 1 (FMR1) gene compared to age matched healthy controls. I did this using dual-task gait and balance paradigms involving i-SWAY and i-WALK inertial sensor testing in conjunction with neuropsychological assessments of executive function. My results showed no significant differences in dual-task interference between the two groups, likely due to low sample size. However, I am continuing to collect data in order to increase my sample size.

Salpi Siyahian

A Controlled Trial of Sertraline in Young Children with Fragile X Syndrome

Affiliation: University of California, Davis

Mentor: Dr. Randy Hagerman

Project Summary: I had the honor of working at the UC Davis MIND Institute as a research study coordinator under the mentorship of Dr. Randi Hagerman. The study I helped coordinate looked at the benefits of a low dose of sertraline, an SSRI that increases the level of serotonin, on language, anxiety, and social deficits in young children with Fragile X syndrome (FXS).

A preliminary analysis of the first 30 subjects showed improvements in cognition, language, and social behavior through significant improvements in the CGI-I, VAS, and Mullen assessments. Working on this project has reinforced my interest in targeted treatments in FXS and I am excited to see the positive impact of this study on the younger FXS population.

This study will continue to enroll children with FXS between the ages of 24-68 months until mid-January 2015.

Matthew Davenport

Excitatory/Inhibitory Modulation and the Fragile X Synapses

Affiliation: Cincinnati Children’s Hospital Medical Center

Mentor: Dr. Craig A. Erickson

Project Summary: I worked with Drs. Craig Erickson and Tori Schaefer to assess the efficacy of two GABA(A) modulators, which boost inhibitory signaling in the FMR1 knock-out mouse model of FXS. Deficiencies in inhibitory signaling, dendritic spine number and spine morphology are suspected to contribute to the pathophysiology of FXS in both patients with FXS and in mouse models.

My work included the chronic treatment of FMR1 knock-out mice, collection and processing of brain tissue, microscopy and quantification of dendritic spine density and characterization of spine morphology.

Preliminary data indicate a differential effect of the therapeutics on the FXS dendritic spine phenotype, likely related to the different specificity of the drugs to regulate inhibitory signaling. Further studies evaluating the effects of these drugs on neural physiology and behavior are currently ongoing.

Whitney Espinel

Improving Health Education for Women Who Carry an FMR1 Premutation

Affiliation: Emory University School of Medicine

Mentor: Dr. Stephanie Sherman

Project Summary: Much of the current research on the FX premutation is focused on defining health risks (e.g., FXPOI/FXTAS) and less is dedicated to understanding the personal experiences of women seeking to understand and navigate their own health journey.

My project used focus group discussions to uncover both barriers and facilitators faced by women who carry the premutation when seeking medical care. We uncovered many barriers to personal healthcare including the lack of knowledge among medical providers, inability to keep pace with findings from research in the field, and the shortage of premutation-specific educational materials and support.

The second half of this project aims to create and distribute premutation-specific educational materials for women. Many premutation carriers face the task of educating not only themselves but also healthcare providers and family members. Having access to up-to-date materials can help diminish misperceptions regarding health risks and aid in information sharing and awareness.

Stefan Sweha

Genetic Markers Predictive of Sertraline Treatment Response in Young Children with Fragile X Syndrome

Affiliation: Department of Biochemistry and Molecular Medicine, University of California, Davis

Mentor: Dr. Flora Tassone

Project Summary: Young children with FXS can present with anxiety, irritability, and hyperactivity related to sensory hyperarousal and language delay. Recently, Winarni et al. (2012) reported that treatment with Sertraline led to improvement in expressive language capability in boys with FXS. A positive response to sertraline was recently observed (Winarni et al., 2010) in young children with FXS and autism spectrum disorders. In addition, a preliminary analysis of children in a double-blind, placebo-controlled trial of sertraline in young children with FXS showed a marked improvement in anxiety.

In this study we determined if the genetic allelic variants of several genes related to the serotonergic pathway correlate with the observed clinical response. Data analysis is currently in progress.

Winarni TI, Chonchaiya W, Adams E, Au J, Mu Y, Rivera SM, Nguyen DV, Hagerman RJ. (2012) Sertraline may improve language developmental trajectory in young children with Fragile X syndrome: a retrospective chart review. Autism ResTreat. 104317. doi: 10.1155/2012/104317.

Ben King

Affiliation: Stanford University

Mentor: Dr. Allan Reiss

Project Summary: This summer, I worked with CIBSR at Stanford University under director Dr. Allan Reiss and project manager Mai Manchanda on a double-blind, placebo-controlled trial of donepezil in Fragile X syndrome (FXS). My work included administering and scoring behavioral and cognitive outcome measures to study participants with a focus on the CNT as well as aiding with the collection of neuroimaging data, including structural and functional MRI and near infrared spectroscopy (NIRS). Furthermore, I carried out structural MRI data editing and quality checking using the brain imaging software FreeSurfer 5.3, a semi-automated anatomical parcellation and segmentation tool.

Based on preliminary analyses of CNT data, donepezil had no effect on cognition in FraX. However, our study utilized many other behavioral/cognitive outcome measures beyond the CNT that must be analyzed to further determine whether donepezil enhances cognition or behavior in FraX.

Liqi Shu

Affiliation: Emory University School of Medicine

Mentor: Peng Jin

Project Summary: Given the high prevalence of Fragile X premutation carriers among the general population and the high risk of developing FXTAS among the male carriers, it is important to develop therapeutic intervention for FXTAS. Using our established assay, we screened multiple libraries, and identified and validated 34 small molecules. Some of them have the unknown biological activities while others have been implicated in different biological pathways, which will be further tested in the mammalian system in the near future. Our studies should provide new insight on the therapeutic development for FXTAS.

Hayle McCausland

Project Summary: Fragile X syndrome is characterized by hyperactivity, and about 25% of patients have epilepsy. FMRP, the protein lost in Fragile X, regulates many different functions in nerve cells. One of its targets is Kv4.2, a protein that is critical for the activity of brain cells. Abnormal levels of Kv4.2 in FXS patients may contribute to the cause of epilepsy. My project was to determine how Kv4.2 is regulated by FMRP. The results suggest a new model for the regulation of Kv4.2 by FMRP.

Zaynah Sadiq

Quality of Life as an Outcome of Developmental Trajectories in Fragile X During the Transition from Adolescence to Adulthood

Affiliation: Stanford University School of Medicine

Michelle Frazer

Correction of Behavioral Phenotypes in FXTAS Model Mice

Affiliation: University of Michigan

Cynthia Chen

Structural Basis for the Recognition of RNA by the Fragile X Mental Retardation Protein, FMRP

Affiliation: Massachusetts Institute of Technology

Scott Danielson

Targeting Excess PI3K Signaling to Rescue Dendritic Spine Morphology in a FXS Mouse Model

Affiliation: Emory University School of Medicine

Annie Pally

FMRP Regulation of the Central Mevalonate Pathway in a Fragile X Mouse Model

Affiliation: University of Houston

Emily Liang

Evaluating FreeSurfer as an Analysis Tool for Longitudinal Structural MRI in Young Adults with FXS

Affiliation: Stanford University