Carolina I. Smith, M.S.

@article{Allard2008,

title = {In vitro cellular adaptations of indicators of longevity in response to treatment with serum collected from humans on calorie restricted diets.},

author = {Joanne S Allard and Leonie K Heilbronn and Carolina Smith and Nicole D Hunt and Donald K Ingram and Eric Ravussin and Rafael de Cabo},

url = {https://www.ncbi.nlm.nih.gov/pubmed/18791640},

doi = {10.1371/journal.pone.0003211},

issn = {1932-6203 (Electronic); 1932-6203 (Linking)},

year  = {2008},

date = {2008-09-15},

journal = {PLoS One},

volume = {3},

number = {9},

pages = {e3211},

address = {Laboratory of Experimental Gerontology, National Institute on Aging, National Institutes of Health, Baltimore, Maryland, United States of America.},

abstract = {Calorie restriction (CR) produces several health benefits and increases lifespan in many species. Studies suggest that alternate-day fasting (ADF) and exercise can also provide these benefits. Whether CR results in lifespan extension in humans is not known and a direct investigation is not feasible. However, phenotypes observed in CR animals when compared to ad libitum fed (AL) animals, including increased stress resistance and changes in protein expression, can be simulated in cells cultured with media supplemented with blood serum from CR and AL animals. Two pilot studies were undertaken to examine the effects of ADF and CR on indicators of health and longevity in humans. In this study, we used sera collected from those studies to culture human hepatoma cells and assessed the effects on growth, stress resistance and gene expression. Cells cultured in serum collected at the end of the dieting period were compared to cells cultured in serum collected at baseline (before the dieting period). Cells cultured in serum from ADF participants, showed a 20% increase in Sirt1 protein which correlated with reduced triglyceride levels. ADF serum also induced a 9% decrease in proliferation and a 25% increase in heat resistance. Cells cultured in serum from CR participants induced an increase in Sirt1 protein levels by 17% and a 30% increase in PGC-1alpha mRNA levels. This first in vitro study utilizing human serum to examine effects on markers of health and longevity in cultured cells resulted in increased stress resistance and an up-regulation of genes proposed to be indicators of increased longevity. The use of this in vitro technique may be helpful for predicting the potential of CR, ADF and other dietary manipulations to affect markers of longevity in humans.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Fischer2007,

title = {Expression of a truncated cystic fibrosis transmembrane conductance regulator with an AAV5-pseudotyped vector in primates.},

author = {Anne C Fischer and Carolina I Smith and Liudmila Cebotaru and Xuemei Zhang and Frederic B Askin and Jerry Wright and Sandra E Guggino and Robert J Adams and Terence Flotte and William B Guggino},

url = {https://www.ncbi.nlm.nih.gov/pubmed/17299412},

doi = {10.1038/sj.mt.6300059},

issn = {1525-0016 (Print); 1525-0016 (Linking)},

year  = {2007},

date = {2007-02-13},

journal = {Mol Ther},

volume = {15},

number = {4},

pages = {756--763},

address = {Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.},

abstract = {Gene therapy using recombinant adeno-associated virus (rAAV2) vectors for cystic fibrosis has shown gene transfer and remarkable safety, yet indeterminate expression. A new construct has been characterized with a powerful exogenous promoter, the cytomegalovirus enhancer/chicken beta-actin promoter, driving a truncated CF transmembrane conductance regulator (CFTR), pseudotyped in an AAV5 viral coat. Our goal is to demonstrate that airway delivery of a pseudotyped rAAV5 vector results in gene transfer as well as expression in non-human primates. Aerosolized pseudotyped rAAV5-DeltaCFTR or rAAV5-GFP (green fluorescent protein) genes were delivered to four and six lungs, respectively. The pseudotyped rAAV5 vector did result in GFP gene transfer (1.005x10(6) copies/mug DNA on average) and quantifiable gene expression. Microscopy confirmed protein expression in airway epithelium. Similarly, the vector also resulted in vector-specific CFTR DNA (1.24x10(5) copies/microg) and mRNA expression. Immunoprecipitation and (32)P phosphoimaging were used to demonstrate CFTR protein expression, as qualitatively enhanced beyond the barely detectable endogenous expression in untreated animals. Based on these promising studies, this CFTR minigene construct is a therapeutic candidate.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Liu2006,

title = {Biological Differences in rAAV Transduction of Airway Epithelia in Humans and in Old World Non-human Primates.},

author = {Xiaoming Liu and Meihui Luo and Cyndi Trygg and Ziying Yan and Diana C M Lei-Butters and Carolina I Smith and Anne C Fischer and Keith Munson and William B Guggino and Bruce A Bunnell and John F Engelhardt},

url = {https://www.ncbi.nlm.nih.gov/pubmed/17667945},

doi = {10.1038/sj.mt.6300277},

issn = {1525-0016 (Print); 1525-0016 (Linking)},

year  = {2006},

date = {2006-07-31},

journal = {Mol Ther},

volume = {15},

number = {12},

pages = {2114--2123},

address = {Department of Anatomy and Cell Biology, College of Medicine, The University of Iowa, Iowa City, Iowa 52242, USA.},

abstract = {Non-human primates (NHPs) are considered to be among the most relevant animal models for pre-clinical testing of human therapies, on the basis of their close evolutionary relatedness to humans in terms of organ cell biology and physiology. In this study, we sought to investigate whether NHP models accurately reflect the effectiveness of recombinant adeno-associated virus (rAAV)-mediated gene delivery to the airway in humans. In order to do this, we utilized an identical model system of differentiated airway epithelia from Indian Rhesus monkeys and from humans, cultured at an air-liquid interface (ALI). In addition to assessing the biology of rAAV-mediated transduction for three serotypes, we characterized the bioelectric properties as a reference for biological similarities and differences between the cell cultures from the two species. Our results demonstrate that airway epithelia from NHPs and humans have very similar Na(+) and Cl(-) transport properties. In contrast, rAAV transduction of airway epithelia of NHPs demonstrated significant differences to those in humans with regard to the efficiency of apical and/or basal transduction with three rAAV serotypes (AAV1, AAV2, AAV5). These findings suggest that the IndianRhesusmonkey may not be the best model for preclinical testing of rAAV-mediated gene therapy to the airway in humans.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Beck2002,

title = {Deposition and expression of aerosolized rAAV vectors in the lungs of Rhesus macaques.},

author = {Suzanne E Beck and Beth L Laube and Carolina I Barberena and Anne C Fischer and Robert J Adams and Kye Chesnut and Terence R Flotte and William B Guggino},

url = {https://www.ncbi.nlm.nih.gov/pubmed/12387250},

issn = {1525-0016 (Print); 1525-0016 (Linking)},

year  = {2002},

date = {2002-10-01},

journal = {Mol Ther},

volume = {6},

number = {4},

pages = {546--554},

address = {Eudowood Division of Pediatric Respitarory Sciences and Departments of Pediatrics, and Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.},

abstract = {The goals of these experiments were to efficiently deliver aerosolized adeno-associated virus (AAV) vector to the lungs of Rhesus macaques and to measure gene transfer and expression. To determine optimal lung deposition, we compared four techniques of delivering aerosolized saline admixed with the radioisotope (99m)technetium ((99m)Tc) nebulized through a mouthpiece (Neb Oral), a laryngeal airway mask (Neb LMA), or an endotracheal tube (Neb ETT), or bronchoscopically delivered by Microsprayer (PennCentury). Total lung deposition fraction, as indicated by gamma scintigraphy, averaged 0.5% (Neb Oral), 1.2% (Neb LMA), 1.8+/-0.4% (Neb ETT), and 62.3+/-11.3% (Microsprayer). Because microspraying was the most efficient method of delivery, we used it to administer saline with (99m)Tc-labeled diethylene-triamine penta-acetic acid (DTPA) admixed with 9 x 10(11) infectious units (i.u.) of AAV serotype 2 (rAAV2) vector encoding green fluorescent protein (GFP; rAAV2-GFP). Initial total and regional lung depositions were quantified by scintigraphy. We analyzed the tissue three weeks later for vector-specific DNA transduction and RNA expression. Radioisotope was detected in all lung regions, reflecting an average dose of 1.33 x 10(10)+/-9.5 x 10(9) i.u. per region. Regional data indicated an increase in expression when the dose exceeded 3 x 10(9) i.u. (P=0.030). We conclude that expression of rAAV2-GFP in lungs appears to be related to depositing a regional threshold dose greater than 3 x 10(9) i.u., easily achieved by bronchoscopic microspraying.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}