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Open access publications by faculty, staff, postdocs, and graduate students in the Department of Medical & Molecular Sciences.
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Item Protein modeling and in silico analysis to assess pathogenicity of ABCA4 variants in patients with inherited retinal disease(Molecular Vision, 2023-10-25) Cevik, Senem; Wangtiraumnuay, Nutsuchar; Van Schelvergem, Kristof; Tsukikawa, Mai; Capasso, Jenina; Biswas, Subhasis B.; Bodt, Barry; Levin, Alex V.; Biswas-Fiss, EstherPurpose: The retina-specific ABCA transporter, ABCA4, plays an essential role in translocating retinoids required by the visual cycle. ABCA4 genetic variants are known to cause a wide range of inherited retinal disorders, including Stargardt disease and cone-rod dystrophy. More than 1,400 ABCA4 missense variants have been identified; however, more than half of these remain variants of uncertain significance (VUS). The purpose of this study was to employ a predictive strategy to assess the pathogenicity of ABCA4 variants in inherited retinal diseases using protein modeling and computational approaches. Methods: We studied 13 clinically well-defined patients with ABCA4 retinopathies and identified the presence of 10 missense variants, including one novel variant in the ABCA4 gene, by next-generation sequencing (NGS). All variants were structurally analyzed using AlphaFold2 models and existing experimental structures of human ABCA4 protein. The results of these analyses were compared with patient clinical presentations to test the effectiveness of the methods employed in predicting variant pathogenicity. Results: We conducted a phenotype–genotype comparison of 13 genetically and phenotypically well-defined retinal disease patients. The in silico protein structure analyses we employed successfully detected the deleterious effect of missense variants found in this affected patient cohort. Our study provides American College of Medical Genetics and Genomics (ACMG)–defined supporting evidence of the pathogenicity of nine missense ABCA4 variants, aligning with the observed clinical phenotypes in this cohort. Conclusions: In this report, we describe a systematic approach to predicting the pathogenicity of ABCA4 variants by means of three-dimensional (3D) protein modeling and in silico structure analysis. Our results demonstrate concordance between disease severity and structural changes in protein models induced by genetic variations. Furthermore, the present study suggests that in silico protein structure analysis can be used as a predictor of pathogenicity and may facilitate the assessment of genetic VUS.Item RNA Landscapes of Brain and Brain-Derived Extracellular Vesicles in Simian Immunodeficiency Virus Infection and Central Nervous System Pathology(The Journal of Infectious Diseases, 2023-12-11) Huang, Yiyao; Abdelgawad, Ahmed; Turchinovich, Andrey; Queen, Suzanne; Abreu, Celina Monteiro; Zhu, Xianming; Batish, Mona; Zheng, Lei; Witwer, Kenneth W.Background Brain tissue-derived extracellular vesicles (bdEVs) act locally in the central nervous system (CNS) and may indicate molecular mechanisms in human immunodeficiency virus (HIV) CNS pathology. Using brain homogenate (BH) and bdEVs from a simian immunodeficiency virus (SIV) model of HIV disease, we identified RNA networks in SIV infection and neuroinflammation. Methods Postmortem occipital cortex samples were obtained from uninfected controls and SIV-infected subjects (acute and chronic phases with or without CNS pathology [SIV encephalitis]). bdEVs were separated and characterized per international consensus guidelines. RNAs from bdEVs and BH were sequenced and quantitative polymerase chain reaction (qPCR)-amplified to detect levels of small RNAs (sRNAs, including microRNAs [miRNAs]) and longer RNAs including messenger RNAs (mRNAs) and circular RNAs (circRNAs). Results Dysregulated RNAs in BH and bdEVs were identified in acute and chronic infection with pathology groups, including mRNAs, miRNAs, and circRNAs. Most dysregulated mRNAs in bdEVs reflected dysregulation in source BH. These mRNAs are disproportionately involved in inflammation and immune responses. Based on target prediction, several circRNAs that were differentially abundant in source tissue might be responsible for specific differences in sRNA levels in bdEVs during SIV infection. Conclusions RNA profiling of bdEVs and source tissues reveals potential regulatory networks in SIV infection and SIV-related CNS pathology.Item Antecubital venous endothelial ETB receptor protein expression is preserved with aging in men(American Journal of Physiology - Heart and Circulatory Physiology, 2024-01-01) Tummala, Saumya; Kuczmarski, Andrew V.; Del Vecchio, Angelica R.; Schwab, Allyson I.; Edwards, David G.; Wenner, Megan M.Changes in endothelial function precede the development of cardiovascular disease (CVD). We have previously shown that age-related declines in endothelial function in women are due in part to a reduction in endothelial cell endothelin-B receptor (ETBR) protein expression. However, it is not known if ETBR protein expression changes with aging in men. The purpose of this study was to test the hypothesis that ETBR protein expression is attenuated in older men (OM) compared with younger men (YM). Primary endothelial cells were harvested from the antecubital vein of 14 OM (60 ± 6 yr; 26 ± 3 kg/m2) and 17 YM (24 ± 5 yr; 24 ± 2 kg/m2). Cells were stained with 4′,6-diamidino-2-phenylindole, vascular endothelial cadherin, and ETBR. Images were quantified using immunocytochemistry. Endothelial function was assessed using brachial artery flow-mediated dilation (FMD). Systolic BP was similar (OM, 123 ± 11 vs. YM, 122 ± 10 mmHg) whereas diastolic BP was higher in OM (OM, 77 ± 7 vs. YM, 70 ± 6 mmHg; P < 0.01). Total testosterone was lower in OM (OM, 6.28 ± 4.21 vs. YM, 9.10 ± 2.68 ng/mL; P = 0.03). As expected, FMD was lower in OM (OM, 3.85 ± 1.51 vs. YM, 6.40 ± 2.68%; P < 0.01). However, ETBR protein expression was similar between OM and YM (OM, 0.39 ± 0.17 vs. YM, 0.42 ± 0.17 AU; P = 0.66). These data suggest that ETBR protein expression is not altered with age in men. These findings contrast with our previous data in women and further support sex differences in the endothelin system. NEW & NOTEWORTHY Our laboratory has previously shown that age-related declines in endothelial function are associated with a reduction in endothelial cell ETBR protein expression in women. However, it is unclear if endothelial cell ETBR protein expression is reduced with aging in men. This study demonstrates that endothelial cell ETBR protein expression is preserved with aging in men, and provides additional evidence for sex differences in the endothelin system.Item An Alkaline Foregut Protects Herbivores from Latex in Forage, but Increases Their Susceptibility to Bt Endotoxin(Life, 2023-11-11) Rajan,VidyaAbout 10% of angiosperms, an estimated 20,000 species, produce latex from ubiquitous isoprene precursors. Latex, an aqueous suspension of rubber particles and other compounds, functions as an antifeedant and herbivory deterrent. It is soluble in neutral to alkaline pH, and coagulates in acidic environments. Here, I propose that foregut-fermenting herbivores such as ruminants, kangaroos, sloths, insect larvae, and tadpoles have adapted to latex in forage with the evolution of alkaline anterior digestive chamber(s). However, they consequently become susceptible to the action of Bacillus thuringiensis (Bt) δ-endotoxin and related bioinsecticides which are activated in alkaline environments. By contrast, hindgut-fermenting herbivores, such as horses and rabbits, have acidic anterior digestive chambers, in which latex coagulates and may cause gut blockage, but in which Bt is not activated. The latex-adapted foregut herbivore vs. latex-maladapted hindgut herbivore hypothesis developed in this paper has implications for hindgut-fermenting livestock and zoo animals which may be provided with latex-containing forage that is detrimental to their gut health. Further, ruminants and herbivorous tadpoles with alkaline anterior chambers are at risk of damage by the supposedly “environmentally friendly” Bt bioinsecticide, which is widely disseminated or engineered into crops which may enter animal feed streams.Item Structural and Pathogenic Impacts of ABCA4 Variants in Retinal Degenerations—An In-Silico Study(International Journal of Molecular Sciences, 2023-04-14) Cevik, Senem; Biswas, Subhasis B.; Biswas-Fiss, Esther E.The retina-specific ATP-binding cassette transporter protein ABCA4 is responsible for properly continuing the visual cycle by removing toxic retinoid byproducts of phototransduction. Functional impairment caused by ABCA4 sequence variations is the leading cause of autosomal recessive inherited retinal disorders, including Stargardt disease, retinitis pigmentosa, and cone-rod dystrophy. To date, more than 3000 ABCA4 genetic variants have been identified, approximately 40 percent of which have not been able to be classified for pathogenicity assessments. This study examined 30 missense ABCA4 variants using AlphaFold2 protein modeling and computational structure analysis for pathogenicity prediction. All variants classified as pathogenic (n = 10) were found to have deleterious structural consequences. Eight of the ten benign variants were structurally neutral, while the remaining two resulted in mild structural changes. This study’s results provided multiple lines of computational pathogenicity evidence for eight ABCA4 variants of uncertain clinical significance. Overall, in silico analyses of ABCA4 can provide a valuable tool for understanding the molecular mechanisms of retinal degeneration and their pathogenic impact.Item Correlations with Congenital Syphilis in the United States: A Cross-Sectional Study(International Journal of Sexually Transmitted Diseases, 2023-07-18) Hughes, Virginia C.Syphilis is caused by the bacterium Treponema pallidum and is transmitted from human to human through sexual contact. Congenital syphilis (CS) occurs when the mother transmits the infection to the fetus. Clinical manifestations of CS include anemia, hepatosplenomegaly, blindness, deafness, meningitis, and deformities in bone structure. The number of cases of CS have increased over the past decade in the United States according to the CDC. A study was conducted correlating the number of Medicaid enrollees in 2020, the number of uninsured persons in 2020, and the number of cases of COVID-19 in 2020 to cases of CS in the United States in 2021. A Spearman rank correlation analysis was done using SPSS. Results were statistically significant for all three pairs of variables with positive correlations; Medicaid enrollment and CS cases (r = 0.735, P<.05), uninsured persons with CS cases (r = 0.713, P<.05), COVID-19 cases and CS cases (r = 0.689, P<.05). Reasons for the increase in CS cases are multifactorial, including variations in state laws regarding syphilis screening in the prenatal period, differences in provider processes for persons on Medicaid, persons uninsured, and restrictions to accessing healthcare providers during the COVID-19 pandemic. Future studies should include questionnaires and interviews with women on their experiences during prenatal visits in regards to syphilis screening, particularly women covered by Medicaid, and surveys completed by healthcare providers to gain insight and to identify factors that affect a woman not being tested for syphilis during her pregnancy.Item Human papillomavirus spectrum of HPV-infected women in Nigeria: an analysis by next-generation sequencing and type-specific PCR(Virology Journal, 2023-07-11) Dom-Chima, Ngozi; Ajang, Yakubu Abubakar; Dom-Chima, Chinyere Ifeoma; Biswas-Fiss, Esther; Aminu, Maryam; Biswas, Subhasis B.Background Human papillomavirus (HPV) infection and cervical cancer are leading health problems and causes of death in many parts of the world. There are ~ 200 HPV types that can infect humans. This study aims to understand the spectrum of HPV infections in Nigerian women with normal or abnormal cytology. Methods We screened cervical samples from 90 women with possible HPV infections collected in two regional hospitals in Nigeria. The first screening was done using next-generation DNA sequencing (NGS), identifying multiple HPV types in many samples. Thereafter, type-specific PCR analysis was used to verify the NGS-identified HPV types in each sample. Results NGS analysis of the 90 samples from the Nigerian cohort identified 44 HPV types. The type-specific PCR confirmed 25 HPV types out of the 44 HPV types detected by NGS, and ~ 10 of these types were the most prevalent. The top five prevalent types found in the Nigerian cohort were HPV71 (17%), HPV82 (15%), HPV16 (16%), HPV6 (10%), and HPV20 (7%). Among the PCR-confirmed HPV types, we found 40.98% high-risk HPV types, 27.22% low-risk HPV types, and 31.15% undetermined HPV types. Among these 25 HPV types in Nigeria, only six were included in the current nine-valent HPV vaccine. We also observed strikingly high multiple HPV infections in most patients, with as many as nine HPV types in a few single samples. Conclusions Our NGS-PCR approach of HPV typing in the Nigerian cohort samples unveiled all possible HPV types currently circulating in Nigerian people. We confirmed 25 HPV types using NGS and PCR, with many samples infected with multiple HPV types. However, only six of these types are part of the nine-valent HPV vaccines indicating the need to develop region-specific selective vaccines.Item Protein–DNA Interactions Regulate Human Papillomavirus DNA Replication, Transcription, and Oncogenesis(International Journal of Molecular Sciences, 2023-05-09) Evande, Roxanne; Rana, Anshul; Biswas-Fiss, Esther E.; Biswas, Subhasis B.Human papillomavirus (HPV) is a group of alpha papillomaviruses that cause various illnesses, including cancer. There are more than 160 types of HPV, with many being “high-risk” types that have been clinically linked to cervical and other types of cancer. “Low-risk” types of HPV cause less severe conditions, such as genital warts. Over the past few decades, numerous studies have shed light on how HPV induces carcinogenesis. The HPV genome is a circular double-stranded DNA molecule that is approximately 8 kilobases in size. Replication of this genome is strictly regulated and requires two virus-encoded proteins, E1 and E2. E1 is a DNA helicase that is necessary for replisome assembly and replication of the HPV genome. On the other hand, E2 is responsible for initiating DNA replication and regulating the transcription of HPV-encoded genes, most importantly the E6 and E7 oncogenes. This article explores the genetic characteristics of high-risk HPV types, the roles of HPV-encoded proteins in HPV DNA replication, the regulation of transcription of E6 and E7 oncogenes, and the development of oncogenesis.Item Enhanced myogenesis through lncFAM-mediated recruitment of HNRNPL to the MYBPC2 promoter(Nucleic Acids Research, 2022-12-19) Chang, Ming-Wen Chang; Yang, Jen-Hao; Tsitsipatis, Dimitrios; Yang, Xiaoling; Martindale, Jennifer L.; Munk, Rachel; Pandey, Poonam R.; Banskota, Nirad; Romero, Brigette; Batish, Mona; Piao, Yulan; Mazan-Mamczarz, Krystyna; De, Supriyo; Abdelmohsen, Kotb; Wilson, Gerald M.; Gorospe, MyriamThe mammalian transcriptome comprises a vast family of long noncoding (lnc)RNAs implicated in physiologic processes such as myogenesis, through which muscle forms during embryonic development and regenerates in the adult. However, the specific molecular mechanisms by which lncRNAs regulate human myogenesis are poorly understood. Here, we identified a novel muscle-specific lncRNA, lncFAM71E1-2:2 (lncFAM), which increased robustly during early human myogenesis. Overexpression of lncFAM promoted differentiation of human myoblasts into myotubes, while silencing lncFAM suppressed this process. As lncFAM resides in the nucleus, chromatin isolation by RNA purification followed by mass spectrometry (ChIRP-MS) analysis was employed to identify the molecular mechanisms whereby it might promote myogenesis. Analysis of lncFAM-interacting proteins revealed that lncFAM recruited the RNA-binding protein HNRNPL to the promoter of MYBPC2, in turn increasing MYBPC2 mRNA transcription and enhancing production of the myogenic protein MYBPC2. These results highlight a mechanism whereby a novel ribonucleoprotein complex, lncFAM-HNRNPL, elevates MYBPC2 expression transcriptionally to promote myogenesis.Item DNA Methylation Analysis Reveals Distinct Patterns in Satellite Cell–Derived Myogenic Progenitor Cells of Subjects with Spastic Cerebral Palsy(Journal of Personalized Medicine, 2022-11-30) Robinson, Karyn G.; Marsh, Adam G.; Lee, Stephanie K.; Hicks, Jonathan; Romero, Brigette; Batish, Mona; Crowgey, Erin L.; Shrader, M. Wade; Akins, Robert E.Spastic type cerebral palsy (CP) is a complex neuromuscular disorder that involves altered skeletal muscle microanatomy and growth, but little is known about the mechanisms contributing to muscle pathophysiology and dysfunction. Traditional genomic approaches have provided limited insight regarding disease onset and severity, but recent epigenomic studies indicate that DNA methylation patterns can be altered in CP. Here, we examined whether a diagnosis of spastic CP is associated with intrinsic DNA methylation differences in myoblasts and myotubes derived from muscle resident stem cell populations (satellite cells; SCs). Twelve subjects were enrolled (6 CP; 6 control) with informed consent/assent. Skeletal muscle biopsies were obtained during orthopedic surgeries, and SCs were isolated and cultured to establish patient–specific myoblast cell lines capable of proliferation and differentiation in culture. DNA methylation analyses indicated significant differences at 525 individual CpG sites in proliferating SC–derived myoblasts (MB) and 1774 CpG sites in differentiating SC–derived myotubes (MT). Of these, 79 CpG sites were common in both culture types. The distribution of differentially methylated 1 Mbp chromosomal segments indicated distinct regional hypo– and hyper–methylation patterns, and significant enrichment of differentially methylated sites on chromosomes 12, 13, 14, 15, 18, and 20. Average methylation load across 2000 bp regions flanking transcriptional start sites was significantly different in 3 genes in MBs, and 10 genes in MTs. SC derived MBs isolated from study participants with spastic CP exhibited fundamental differences in DNA methylation compared to controls at multiple levels of organization that may reveal new targets for studies of mechanisms contributing to muscle dysregulation in spastic CP.Item The Impact of Cancer Cases, Marijuana possession Arrests, and Opioid Deaths on Cannabis Policies in the United States: A Logistic Regression Study(American Journal of Public Health Research, 2022-09-14) Hughes, Virginia C.Recreational cannabis is currently legal in nineteen states and the District of Columbia. The history of each states pathway for passing laws codifying fully legal status varies greatly across the United States. A study was conducted with the aim of identifying factors that significantly impact a states fully legal status on cannabis employing a logistic regression design. Independent factors analyzed included the marijuana possession arrest rate (MPAR), new cancer cases, and opioid overdose rate. All data were from 2010 to assess if these factors impacted passage of laws approving recreational cannabis, as all such laws were passed after 2010. The dependent variable was dichotomous toward fully legal status or not fully legal status in states. Results showed statistically significance with the MPAR variable (P<.05). The opioid over dose rate and cancer cases did not yield statistically significant results. Consistent with the federalist system, select state legislatures have made the decision to pass laws regarding recreational cannabis propelled by public support when cannabis is still illegal under federal law. This paper delineates both recreational cannabis and medical cannabis laws, and provides salient discussion on variables analyzed and ideas for future policy studies.Item The Medical Diagnostics Major: Gateway to Medical School and Physician Assistant School(American Journal of Educational Research, 2022-05-25) Hughes, Virginia C.Medical diagnostics, offered at the University of Delaware College of Health Sciences, represents a novel major in higher education. This major incorporates courses similar to the preclinical content of professional schools along with the standard science and mathematics prerequisites. Students have the option of taking a pre-physician assistant (PA) concentration or non-pre-PA concentration. Students in the non-pre-PA concentration typically apply to medical or dental school, while students in the pre-PA concentration apply to PA school. The medical diagnostics major began in 2013 in a 2+2 format. Students completed prerequisite courses in their first two years and then applied to the permanent major, medical diagnostics. In 2021, the format for both concentrations transitioned to a direct admissions model. A study was conducted with the goal of ascertaining if there was an increase or decrease in enrollment from 2020 to 2021. Results of the study indicated a ten percent increase in enrollment for Medical Diagnostics pre -PA and a nineteen percent decrease in enrollment for Medical Diagnostics non-pre PA. This paper outlines the critical role of the pre-health advisor for students who plan to apply to medical school or PA school as well as administrative procedures associated with a 2+2 model compared to a direct admissions model.Item Insertional Mutagenesis by CRISPR/Cas9 Ribonucleoprotein Gene Editing in Cells Targeted for Point Mutation Repair Directed by Short Single-Stranded DNA Oligonucleotides(Public Library of Science, 2017-01-04) Rivera-Torres, Natalia; Banas, Kelly; Bialk, Pawel; Bloh, Kevin M.; Kmiec, Eric B.; Kelly Banas; Pawel Bialk; Kevin M. Bloh; Eric B. Kmiec; Natalia Rivera-Torres; Kmiec, EricCRISPR/Cas9 and single-stranded DNA oligonucleotides (ssODNs) have been used to direct the repair of a single base mutation in human genes. Here, we examine a method designed to increase the precision of RNA guided genome editing in human cells by utilizing a CRISPR/Cas9 ribonucleoprotein (RNP) complex to initiate DNA cleavage. The RNP is assembled in vitro and induces a double stranded break at a specific site surrounding the mutant base designated for correction by the ssODN. We use an integrated mutant eGFP gene, bearing a single base change rendering the expressed protein nonfunctional, as a single copy target in HCT 116 cells. We observe significant gene correction activity of the mutant base, promoted by the RNP and single-stranded DNA oligonucleotide with validation through genotypic and phenotypic readout. We demonstrate that all individual components must be present to obtain successful gene editing. Importantly, we examine the genotype of individually sorted corrected and uncorrected clonally expanded cell populations for the mutagenic footprint left by the action of these gene editing tools. While the DNA sequence of the corrected population is exact with no adjacent sequence modification, the uncorrected population exhibits heterogeneous mutagenicity with a wide variety of deletions and insertions surrounding the target site. We designate this type of DNA aberration as on-site mutagenicity. Analyses of two clonal populations bearing specific DNA insertions surrounding the target site, indicate that point mutation repair has occurred at the level of the gene. The phenotype, however, is not rescued because a section of the single-stranded oligonucleotide has been inserted altering the reading frame and generating truncated proteins. These data illustrate the importance of analysing mutagenicity in uncorrected cells. Our results also form the basis of a simple model for point mutation repair directed by a short single-stranded DNA oligonucleotides and CRISPR/Cas9 ribonucleoprotein complexItem Identifying specific proteins involved in eggshell membrane formation using gene expression analysis and bioinformatics(BioMed Central Ltd., 2015-10-15) Du, Jingwen; Hincke, Maxwell T.; Rose-Martel, Megan; Hennequet-Antier, Christelle; Brionne, Aurelien; Cogburn, Larry A.; Nys, Yves; Gautron, Joel; Jingwen Du, Maxwell T. Hincke, Megan Rose-Martel, Christelle Hennequet-Antier, Aurelien Brionne, Larry A. Cogburn, Yves Nys and Joel Gautron; Cogburn, Larry A.Background The avian eggshell membranes surround the egg white and provide a structural foundation for calcification of the eggshell which is essential for avian reproduction; moreover, it is also a natural biomaterial with many potential industrial and biomedical applications. Due to the insoluble and stable nature of the eggshell membrane fibres, their formation and protein constituents remain poorly characterized. The purpose of this study was to identify genes encoding eggshell membrane proteins, particularly those responsible for its structural features, by analyzing the transcriptome of the white isthmus segment of the oviduct, which is the specialized region responsible for the fabrication of the membrane fibres. Results The Del-Mar 14 K chicken microarray was used to investigate up-regulated expression of transcripts in the white isthmus (WI) compared with the adjacent magnum (Ma) and uterine (Ut) segments of the hen oviduct. Analysis revealed 135 clones hybridizing to over-expressed transcripts (WI/Ma + WI/Ut), and corresponding to 107 NCBI annotated non-redundant Gallus gallus gene IDs. This combined analysis revealed that the structural proteins highly over-expressed in the white isthmus include collagen X (COL10A1), fibrillin-1 (FBN1) and cysteine rich eggshell membrane protein (CREMP). These results validate previous proteomics studies which have identified collagen X (α-1) and CREMP in soluble eggshell extracts. Genes encoding collagen-processing enzymes such as lysyl oxidase homologs 1, 2 and 3 (LOXL1, LOXL2 and LOXL3), prolyl 4 hydroxylase subunit α-2 and beta polypeptide (P4HA2 and P4HB) as well as peptidyl-prolyl cis-trans isomerase C (PPIC) were also over-expressed. Additionally, genes encoding proteins known to regulate disulfide cross-linking, including sulfhydryl oxidase (QSOX1) and thioredoxin (TXN), were identified which suggests that coordinated up-regulation of genes in the white isthmus is associated with eggshell membrane fibre formation. Conclusions The present study has identified genes associated with the processing of collagen, other structural proteins, and disulfide-mediated cross-linking during eggshell membrane formation in the white isthmus. Identification of these genes will provide new insight into eggshell membrane structure and mechanisms of formation that will assist in the development of selection strategies to improve eggshell quality and food safety of the table egg.Item Initial Observations of Cell-Mediated Drug Delivery to the Deep Lung(Cognizant Communication Corporation, 2011) Glaum, M.; Kumar, Arun (Professor); El-Badri, N.; Mohapatra, S.; Haller, E.; Park, Sunhee; Patrick, L.; Nattkemper, L.; Vo, D.; Cameron, D. F.; Kumar, A., Glaum, M., El-Badri, N., Mohapatra, S., Haller, E., Park, S., Patrick, L., Nattkemper, L., Vo, D., Cameron, D. F.; Kumar, Arun (Professor)Using current methodologies, drug delivery to small airways, terminal bronchioles, and alveoli (deep lung) is inefficient, especially to the lower lungs. Urgent lung pathologies such as acute respiratory distress syndrome (ARDS) and post-lung transplantation complications are difficult to treat, in part due to the methodological limitations in targeting the deep lung with high efficiency drug distribution to the site of pathology. To overcome drug delivery limitations inhibiting the optimization of deep lung therapy, isolated rat Sertoli cells preloaded with chitosan nanoparticles were use to obtain a high-density distribution and concentration (92%) of the nanoparticles in the lungs of mice by way of the peripheral venous vasculature rather than the more commonly used pulmonary route. Additionally, Sertoli cells were preloaded with chitosan nanoparticles coupled with the anti-inflammatory compound curcumin and then injected intravenously into control or experimental mice with deep lung inflammation. By 24 h postinjection, most of the curcumin load ( 90%) delivered in the injected Sertoli cells was present and distributed throughout the lungs, including the perialveloar sac area in the lower lungs. This was based on the high-density, positive quantification of both nanoparticles and curcumin in the lungs. There was a marked positive therapeutic effect achieved 24 h following curcumin treatment delivered by this Sertoli cell nanoparticle protocol (SNAP). Results identify a novel and efficient protocol for targeted delivery of drugs to the deep lung mediated by extratesticular Sertoli cells. Utilization of SNAP delivery may optimize drug therapy for conditions such as ARDS, status asthmaticus, pulmonary hypertension, lung cancer, and complications following lung transplantation where the use of high concentrations of anti-inflammatory drugs is desirable, but often limited by risks of systemic drug toxicity.