Pediatrics 2022

Genome Editing for Sickle Cell Disease and β-Thalassemia

Faculty Mentor’s Name: Dr. Arun Srivastava
Email: aruns@peds.ufl.edu
Phone Number: (352) 273-8259
Project Category: Basic
International Component or Travel: No

Research Project Description:

Human hemoglobinopathies, such as sickle cell disease (SCD) and beta-thalassemia, are by far the most common (1 in 600) monogenic diseases worldwide, and are among the likely candidates for their potential treatment provided that pluripotent hematopoietic stem cells (HSCs) can be stably transduced, and long-term, regulated expression of a functional beta-globin gene in the erythroid progenitor cells can be achieved. Although lentiviral vectors have been shown to be effective in animal models, their safety remains to be established since development of acute myeloid leukemia in several patients in clinical trials in patients with beta-thalassemia was reported recently. Thus, we believe that further development of alternative vector systems, such as the adeno-associated virus (AAV), needs to be pursued for their potential ability to achieve high efficiency transduction of HSCs, given the proven safety and efficacy of AAV vectors in clinical trials in gene therapy of Leber’s congenital amaurosis and hemophilia B. We have developed optimized AAV vectors that are capable of high-efficiency transduction in general, and HSCs in particular. Medical students will be involved in using these optimized AAV vectors for “nuclease-free” genome editing in hematopoietic stem cells. Funding for the project is provided by the National Institutes of Health and the George H. Kitzman Endowment.

Tan, K. Qing, S. Zhou, M.C. Yoder, and A. Srivastava. Adeno-associated virus 2-mediated transduction and erythroid lineage-restricted long-term expression of the human b-globin gene in hematopoietic cells from homozygous b-thalassemic mice. Molecular Therapy, 3: 940-946, 2001.

Maina, L. Zhong, X.Li, W. Zhao, Z. Han, D. Bischof, G, Aslanidi, S, Zolotukhin, K. Weigel-Van Aken, A. Rivers, W.B. Slayton, M.C. Yoder, and Srivastava, A. Optimization of recombinant adeno-associated virus serotype vectors for human b-globin gene transfer and transgene expression. Human Gene Therapy, 19: 365-375, 2008.

Song, M.A. Kauss, E. Kopin, M. Chandra, T. Ul-Hasan, E. Miller, G.R. Jayandharan, A.E. Rivers, G.V. Aslanidi, C. Ling, B. Li, W. Ma, X. Li, L.M. Andino, L. Zhong, A.F. Tarantal, M.C.Yoder, K.K. Wong, Jr., M. Tan, S. Chatterjee, and A. Srivastava. Optimizing the transduction efficiency of capsid-modified AAV6 vectors in primary human hematopoietic stem cells in vitro and in a xenograft mouse model in vivo. Cytotherapy, 15: 986-998, 2013.

Song, X. Li, G.R. Jayandharan, Y. Wang, G.V. Aslanidi, C. Ling, L. Zhong, G. Gao, M.C. Yoder, C. Ling, M. Tan, and A. Srivastava. High-efficiency transduction of primary human hematopoietic stem cells and erythroid lineage-restricted expression by optimized AAV6 serotype vectors in vitro and in a murine xenograft model in vivo. PLoS One, 8(3): e58757, 2013.

Ling, K. Bhukhai, Z. Yin, M.Q. Tan, M.C. Yoder, P. Leboulch, E. Payen, and A. Srivastava. High-efficiency transduction of primary human CD34+ hematopoietic stem/progenitor cells by AAV6 serotype vectors: Strategies for overcoming donor-variation and implications in genome editing. Scientific Reports, 6, 35495, 2016.

Yang, K. Qing, G.D. Keeler, L. Yin, M. Mietzsch, C. Ling, B.E. Hoffman, M. Agbandje-McKenna, M. Tan, W. Wang, and A. Srivastava. Enhanced transduction of human hematopoietic stem cells by AAV6 vectors: Implications in gene therapy and genome editing. Molecular Therapy-Nucleic Acids. 20: 451-458, 2020.

Gene Therapy of Human Liver Cancer

Faculty Mentor’s Name: Dr. Arun Srivastava
Email: aruns@peds.ufl.edu
Phone Number: (352) 273-8259
Project Category: Basic
International Component or Travel: No

Research Project Description:

Two of the most common liver cancers include hepatoblastoma (HB) in children, and hepatocellular carcinoma (HCC) in adults. HB is the most frequent pediatric liver cancer in the United States. HCC is one of the five most common cancers involving solid tumors, and is highly fatal. HCC is rapidly emerging as a clinically important disease in developed countries and of grave concern is its increasing incidence to epidemic proportions in certain areas of the world. An alarming trend in its increase has been recently observed: 18,000 new cases of HCC are expected every year. Projected new cases in the coming years are likely to rise even more dramatically, based on the increasing number of hepatitis-C infected Americans (~4 million). Furthermore, the increasing incidence in the United States of obesity and diabetes, which have also been linked to the development of chronic liver disease and HCC, is likely to further augment the number of Americans afflicted with HCC. Thus, a novel therapeutic approach for the treatment of HB and HCC is warranted, which may involve the use of target-specific genes. We have had a long-term interest in a non-pathogenic human parvovirus, the adeno-associated virus (AAV), which has gained attention as a potentially safe vector for gene therapy, and has recently shown efficacy in Phase I clinical trials in patients with Leber’s congenital amaurosis and hemophilia B. We have observed that of the 10 AAV serotypes, AAV3 vectors transduce human HB and HCC cell lines and primary human hepatocytes extremely well, and also target human liver tumors in mouse xenograft models extremely efficiently. Medical students will be involved in using these optimized AAV3 vectors carrying therapeutic genes to potentially eradicate human liver tumors in mouse xenograft models. Funding for the project is provided by the National Institutes of Health and the George H. Kitzman Endowment.

Ling, Y. Lu, B. Cheng, K.E. McGoogan, S.W.Y. Gee, W. Ma, B. Li, G.V. Aslanidi, and A. Srivastava. High-efficiency transduction of liver cancer cells by recombinant adeno-associated virus serotype 3 vectors. J. Vis. Exp., 49. Pii: 2538, doi: 10.3791/2538, 2011.

B. Cheng, C. Ling, Y. Dai, L.G. Glushakova, Y. Lu, S.W.Y. Gee, K.E. McGoogan, G.V. Aslanidi, M. Park, P.W. Stacpoole, D. Siemann, C. Liu, Srivastava, and C. Ling. Development of optimized AAV3 serotype vectors: Mechanism of high-efficiency transduction of human liver cancer cells. Gene Therapy, 19: 375-384, 2012.

Ling, Y. Wang, Y. Zhang, A. Ejjigani, Z. Yin, Y. Lu, L. Wang, M. Wang, J. Li, Z. Hu, G.V. Aslanidi, L. Zhong, G. Gao, A. Srivastava, and C. Ling. Selective in vivo targeting of human liver tumors by optimized recombinant AAV3 vectors in a murine xenograft model. Human Gene Therapy, 25: 1023-1034, 2014.

Zhang, L, Wang, Y. Lu, G.V. Aslanidi, A. Srivastava, C. Ling, and C. Ling. Cytotoxic genes from traditional Chinese medicine inhibit tumor growth both in vitro and in vivo. J. Int. Medicine, 12: 483-494, 2014.

Yin, G.D. Keeler, Y. Zhang, B.E. Hoffman, C. Ling, K. Qing, and A. Srivastava. AAV3-miRNA vectors for growth suppression of human hepatocellular carcinoma cells in vitro and human liver tumors in a murine xenograft model in vivo. Gene Therapy, 7-8:422-434, 2021.

Reliability of spot check transcutaneous hemoglobin measurements in children in the pediatric hematology/oncology clinic

Faculty Mentor’s Name: Dr. Molly Posa
Email: mollyposa@peds.ufl.edu
Phone Number: (352) 222-9688
Project Category: Clinical
International Component or Travel: No

Research Project Description:

The overall incidence of childhood cancer is increasing with over 17,000 children diagnosed each year. A majority of the treatment options require frequent venipunctures to evaluate for and treat anemia. There is a noninvasive transcutaneous hemoglobin test available, but little research has been done to determine the reliability in the pediatric oncology population. Our study assessed the accuracy and reliability of a non-invasive hemoglobin measurement device compared with the gold standard, venous hemoglobin measurement, in the pediatric oncology clinic setting.

Enrollment will include any pediatric oncology/hematology patient who needs a hemoglobin measurement during their pediatric oncology/hematology outpatient appointment during the specified study period. After agreement to participate, each participant will have a hemoglobin measurement obtained with Pronto Pulse Co-Oximeter during the same period as their already ordered venous hemoglobin sample. The venous hemoglobin sample will be obtained by trained medical personnel (nurses or phlebotomists in the hematology/oncology clinic). Paired hemoglobin results will be compared.

The medical student will be responsible for submitting an IRB-1, obtaining informed consent, obtaining the participant transcutaneous hemoglobin measurement in the pediatric oncology/hematology clinic, completing a chart review of pediatric subjects to collect and review data (to be determined by MSRP mentors and student). The correlation between the two measurements (venous vs transcutaneous) will be evaluated on Bland-Altman analysis. Medical students will participate in data collection and analysis of the chart review along with preparation of abstracts and manuscripts resulting from the study. Results of this study will help determine the validity and reliability of a non-invasive hemoglobin measurement device in the pediatric oncology/hematology clinic setting.

Gene Therapy of Sickle Cell Disease and β-Thalassemia

Faculty Mentor’s Name: Dr. Arun Srivastave
Email: aruns@peds.ufl.edu
Phone Number: (352) 283-8259
Project Category: Basic
International Component or Travel: No

Research Project Description:

Human hemoglobinopathies, such as sickle cell disease (SCD) and beta-thalassemia, are by far the most common (1 in 600) monogenic diseases worldwide, and are among the likely candidates for their potential treatment provided that pluripotent hematopoietic stem cells (HSCs) can be stably transduced, and long-term, regulated expression of a functional beta-globin gene in the erythroid progenitor cells can be achieved. Although lentiviral vectors have been shown to be effective in animal models, their safety remains to be established since development of acute myeloid leukemia in several patients in clinical trials in patients with beta-thalassemia was reported recently. Thus, we believe that further development of alternative vector systems, such as the adeno-associated virus (AAV), needs to be pursued for their potential ability to achieve high efficiency transduction of HSCs, given the proven safety and efficacy of AAV vectors in clinical trials in gene therapy of Leber’s congenital amaurosis and hemophilia B. We have developed optimized AAV vectors that are capable of high-efficiency transduction in general, and HSCs in particular. Students will be involved in using these optimized AAV vectors for “nuclease-free” genome editing in hematopoietic stem cells. Funding for the project is provided by the National Institutes of Health and the George H. Kitzman Endowment.

Gene Therapy of Human Liver Cancer

Faculty Mentor’s Name: Dr. Arun Srivastave
Email: aruns@peds.ufl.edu
Phone Number: (352) 283-8259
Project Category: Basic
International Component or Travel: No

Research Project Description:

Two of the most common liver cancers include hepatoblastoma (HB) in children, and hepatocellular carcinoma (HCC) in adults. HB is the most frequent pediatric liver cancer in the United States. HCC is one of the five most common cancers involving solid tumors, and is highly fatal. HCC is rapidly emerging as a clinically important disease in developed countries and of grave concern is its increasing incidence to epidemic proportions in certain areas of the world. An alarming trend in its increase has been recently observed: 18,000 new cases of HCC are expected every year. Projected new cases in the coming years are likely to rise even more dramatically, based on the increasing number of hepatitis-C infected Americans (~4 million). Furthermore, the increasing incidence in the United States of obesity and diabetes, which have also been linked to the development of chronic liver disease and HCC, is likely to further augment the number of Americans afflicted with HCC. Thus, a novel therapeutic approach for the treatment of HB and HCC is warranted, which may involve the use of target-specific genes. We have had a long-term interest in a non-pathogenic human parvovirus, the adeno-associated virus (AAV), which has gained attention as a potentially safe vector for gene therapy, and has recently shown efficacy in Phase I clinical trials in patients with Leber’s congenital amaurosis and hemophilia B. We have observed that of the 10 AAV serotypes, AAV3 vectors transduce human HB and HCC cell lines and primary human hepatocytes extremely well, and also target human liver tumors in mouse xenograft models extremely efficiently. Students will be involved in using these optimized AAV3 vectors carrying therapeutic genes to potentially eradicate human liver tumors in mouse xenograft models. Funding for the project is provided by the National Institutes of Health and the George H. Kitzman Endowment.

Ling, Y. Lu, B. Cheng, K.E. McGoogan, S.W.Y. Gee, W. Ma, B. Li, G.V. Aslanidi, and A. Srivastava. High-efficiency transduction of liver cancer cells by recombinant adeno-associated virus serotype 3 vectors. J. Vis. Exp., 49. Pii: 2538, doi: 10.3791/2538, 2011.

B. Cheng, C. Ling, Y. Dai, L.G. Glushakova, Y. Lu, S.W.Y. Gee, K.E. McGoogan, G.V. Aslanidi, M. Park, P.W. Stacpoole, D. Siemann, C. Liu, Srivastava, and C. Ling. Development of optimized AAV3 serotype vectors: Mechanism of high-efficiency transduction of human liver cancer cells. Gene Therapy, 19: 375-384, 2012.

Ling, Y. Wang, Y. Zhang, A. Ejjigani, Z. Yin, Y. Lu, L. Wang, M. Wang, J. Li, Z. Hu, G.V. Aslanidi, L. Zhong, G. Gao, A. Srivastava, and C. Ling. Selective in vivo targeting of human liver tumors by optimized recombinant AAV3 vectors in a murine xenograft model. Human Gene Therapy, 25: 1023-1034, 2014.

Zhang, L, Wang, Y. Lu, G.V. Aslanidi, A. Srivastava, C. Ling, and C. Ling. Cytotoxic genes from traditional Chinese medicine inhibit tumor growth both in vitro and in vivo. J. Int. Medicine, 12: 483-494, 2014.

Yin, G.D. Keeler, Y. Zhang, B.E. Hoffman, C. Ling, K. Qing, and A. Srivastava. AAV3-miRNA vectors for growth suppression of human hepatocellular carcinoma cells in vitro and human liver tumors in a murine xenograft model in vivo. Gene Therapy, 7-8:422-434, 2021.

Determining the Natural History of Exocrine Dysfunction in Pre-Type 1 Diabetes

Faculty Mentor’s Name: Dr. Brittany Bruggeman
Email: bruggemanbr@ufl.edu
Phone Number: (321) 537-8832
Project Category: Translational
International Component or Travel: No

Research Project Description:

Type 1 diabetes is historically thought of as a beta-cell specific disease, however, subclinical exocrine pancreatic dysfunction is frequently present at diagnosis. We hypothesize that a decline in exocrine function occurs prior to the development of clinical type 1 diabetes and that a decline in fecal elastase, the most sensitive and specific clinical marker of exocrine function, can be used to predict the onset of type 1 diabetes. A large biorepository of samples from the TEDDY study (The Environmental Determinants of Diabetes in the Young) will be used to measure longitudinal fecal elastase levels in a large sample of subjects with pre-type 1 diabetes (n=250) to determine the natural history of exocrine dysfunction in this population.

Medical students will be trained to run ELISAs, enter data into REDCap, and will be involved in the analysis and publication of our results. It is expected that this project will result in at least 1-2 publications and that the medical student will be an author on these publications, as long as they participate meaningfully in the work. This project is already funded by the NIH NIDDK, UF CTSI, and Pediatric Endocrine Society.

Exocrine Pancreatic Enzymes as Biomarkers of Response to Immunotherapy in Type 1 Diabetes

Faculty Mentor’s Name: Dr. Brittany Bruggeman
Email: bruggemanbr@ufl.edu
Phone Number: (321) 537-8832
Project Category: Clinical
International Component or Travel: No

Research Project Description:

A differential trend in exocrine pancreatic enzymes in response to type 1 diabetes immunotherapy could improve our ability to predict disease progression and contribute to our understanding of type 1 diabetes pathophysiology. We hypothesize that exocrine pancreatic enzymes (amylase, lipase, and trypsin) will increase in new-onset type 1 diabetes responders to immunotherapy but will decline in those who do not respond and in controls.

This data has already been collected but needs to be compiled, organized in a REDCap database, analyzed, and written up for publication. Medical students will be trained to compile data and will work with a statistician on the analysis of the data and will help to write the paper. It is expected that they will be co-author on this publication if they complete this work. This work has funding for completion of the statistical analysis through the UF CTSI KL2 program.

Transition Readiness in a Pediatric Hematology and Oncology Population

Faculty Mentor’s Name: Dr. Tung Wynn
Email: twynn@ufl.edu
Phone Number: (352) 273-9120
Project Category: CQI
International Component or Travel: No

Research Project Description:

Many pediatric hematology and oncology patients are surviving their diseases and living long into adulthood. The need for these patients to transition from their pediatric providers to appropriate adult providers continues to grow. In the UF pediatric hematology and oncology clinic, patients approaching this transition are asked to answer a Transition Readiness Assessment Questionnaire (TRAQ). The TRAQ is performed as a part of a statewide initiative and how they are used in this project would be of great interest to other centers in Florida. The purpose of this proposed project is to review the responses on the TRAQ and determine the topics and skills that patients feel they are most frequently ill prepared. The project will advise practitioners on which topics need to be discussed during routine visits and what additional educational materials may need to be developed to assist in these discussions.

Adult caregiver COVID vaccination through administration in pediatric outpatient clinics: A cocooning health care improvement project.

Faculty Mentor’s Name: Dr. Maria Kelly
Email: mkelly@ufl.edu
Phone Number: (352) 733-3577
Project Category: Clinical
International Component or Travel: No

Research Project Description:

Pediatric exposure to COVID-infected adult caregivers (AC) is a significant risk factor for developing coronavirus, especially for children at high risk due to chronic illness or those unable to become vaccinated currently (vaccination not approved < 5 years of age). Barriers to vaccination likely include scheduling obstacles, perceived cost and vaccine misinformation. Pediatric COVID disease burden is high in the pediatric population and cocooning of caretakers may be an opportunity to improve caretaker vaccination rates and reduce risk to unvaccinated infants and children. Cocooning has been studied and recommended as a best practice for influenza vaccination so could have the same positive outcomes during the COVID pandemic to improve community vaccination rates and decrease risk to unvaccinated infants and children.

Objective: To explore the intervention of offering COVID vaccination to adult caretakers (ACs) within their child’s pediatric medical home and determine barriers to access to improve future experience.

Design/Methods: Suggest a retrospective chart review at our pediatric primary care sites to identify ACs who received an COVID vaccination at one of 5 UF pediatric primary care clinics. To evaluate the process, each encounter can be screened for demographics of both the AC and their children to best understand the process and uptake of this practice.

95210 Health Tracker in Pediatrics

Faculty Mentor’s Name: Dr. Angelina Bernier
Email: angelina@ufl.edu
Phone Number: (352) 265-7337
Project Category: Clinical
International Component or Travel: No

Research Project Description:

This project is a chart review to examine the trends in health and fitness for a pediatric population followed in the UF Pediatric Metabolic & Obesity clinic. The 95210 Health Tracker is a survey that was created as part of a wellness initiative that encourages participants to reach their personal health and wellness goals by tracking health behaviors such as exercise and nutrition based on America Academy of Pediatric guidelines for obesity prevention. The aim is to determine the wellness behaviors of this specific pediatric population and assess survey response trends over time in subsequent follow up visits.