United States

Background: Hispanic Americans comprise about 25% of the United States population but only 3% of orthopaedic surgeons. Therefore, Spanish-speaking patients facing communication barriers may use tools like large language models (LLMs), such as ChatGPT, for medical information. Given the rising use of LLMs for patient education, it is critical to evaluate whether language-based disparities exist in the quality of information provided. While prior research has evaluated ChatGPT’s performance between languages across various specialties, no study has
examined this in sports surgery. The purpose of this study was to compare the accuracy of ChatGPT responses to patient questions about anterior cruciate ligament reconstruction (ACLR) in English versus Spanish. We hypothesized ChatGPT will provide generally accurate responses overall, but that responses in English will be more accurate.
Methods: Using methodology consistent with prior orthopaedic research, ten commonly asked patient questions related to ACLR were compiled via review of patient education materials from various orthopaedic institutions. Each question was independently submitted to ChatGPT in both English and Spanish using separate chat threads. Responses were evaluated by two
sports medicine fellowship-trained orthopaedic surgeons per language, using a standardized 5-point Likert scale (1 = major errors; 5 = completely accurate). Mean accuracy scores were calculated for each language. Inter-rater reliability was assessed using the intraclass correlation coefficient (ICC[2,k]) for average ratings. A Mann–Whitney U test was performed to compare average accuracy scores between English and Spanish responses.
Results: Inter-rater reliability was acceptable (ICC = 0.727 for both languages). The mean accuracy score was 4.45±0.69 for English responses and 4.65±0.53 for Spanish responses. There was no significant difference in accuracy between the languages (p = 0.460). Qualitatively, raters indicated most responses provided excellent, detailed summaries, but some responses, particularly regarding graft options, provided outdated information.
Conclusion: There was no significant difference in accuracy between English and Spanish responses generated by ChatGPT to patient questions regarding ACLR. This finding is novel, as no prior studies have evaluated language-based differences in ChatGPT accuracy within sports surgery. Given the noted underrepresentation of Hispanic Americans among orthopaedic surgeons in the United States, these results suggest that ChatGPT may be a valid source of information for Spanish-speaking patients facing language barriers when seeking care for ACL injuries. However, regardless of language, some responses appeared to rely on outdated information. As such, LLMs like ChatGPT should continue to be used with caution in clinical practice.

AMA Research Challenge 2025 

Accuracy of English vs. Spanish ChatGPT Responses to Patient Questions
about ACL Reconstruction

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Background

Human papillomaviruses (HPV) cause most cervical carcinomas and substantial fractions of oropharyngeal, anal, vulvar, penile, and vaginal carcinomas. During infection, HPV DNA initially replicates as circular episomes. However, it becomes integrated into the host genome in most HPV-induced cancers. Following integration, HPV is most commonly found as a subgenomic segment that includes the non-coding regulatory region (URR) and viral oncogenes E6 and E7. In tumors, HPV DNA can exist as extrachromosomal DNA (ecDNA) circles, chimeric HPV-human tandem repeats (heterocatemers), or both. The presence of ecDNA is associated with poor patient prognosis across cancer types. However, distinguishing whether particular amplified DNAs in tumors are ecDNAs and/or intrachromosomal heterocatemers and determining the extent of genomic rearrangement surrounding them, present technical challenges. Characterizing HPV-related integration structures accurately and completely is essential for advancing our understanding of HPV-driven tumorigenesis and the mechanisms behind viral integration.

Methods

We applied a battery of genomic analysis techniques at various size-scales to the HPV16-positive human oropharyngeal squamous cell carcinoma cell line UM-SCC-47. These techniques included a custom hybridization capture assay coupled with short-read Illumina sequencing (HC+SEQ), Nanopore long-read DNA and RNA sequencing, Bionano optical genome mapping, and single-cell fluorescence in situ hybridization (FISH). 

Results

HC+SEQ and long-read DNA sequencing defined the structure of the integrated HPV16 DNA, determined the human-virus DNA junctions, and mapped a repeated 23.5 kb HPV-human heterocatemer at single-base-pair resolution. RNA-seq revealed that these heterocatamers could produce spliced, polyadenylated virus-human fusion transcripts alongside high viral oncogene expression. Optical genome mapping further showed that the 23.5 kb heterocatemer occurred in tandem arrays of various lengths, ranging from one unit to more than 27 tandem units. These large-scale arrays (up to 700 kb) were further rearranged with adjacent human DNA in even larger-scale structures. Additionally, single-cell FISH analysis revealed that every cell had intrachromosomal HPV16 associated with human DNA, with a subset of cells also containing HPV16 ecDNA.

Conclusion

These findings highlight extensive genomic instability, complexity, and heterogeneity of HPV16-human DNA structures in this cell line, including previously undescribed higher-order organizational patterns. They also underscore the value of combining large-scale analyses to resolve complex genomic rearrangements and emphasize the importance of FISH for distinguishing intrachromosomal from extrachromosomal DNA structures. Ultimately, understanding how integrated HPV DNA higher-order structures contribute to tumorigenesis could lead to patient stratification based on individual risk of progression or inform the development of targeted therapeutic strategies.

Defining complex and heterogeneous HPV-human DNA structures through large-scale genomic analyses

Background
Molar–incisor hypomineralization (MIH) is a common developmental enamel defect in children, leading to higher risk of caries, post-eruptive enamel breakdown, and dental pain. Proper enamel formation depends on the expression and function of matrix proteins like ameloblastin (Ambn) and the protease matrix metalloproteinase 20 (Mmp20), which regulates mineral deposition and organic matrix removal during amelogenesis. We hypothesized that enamel hypomineralization caused by Ambn overexpression in mice could be rescued by increasing Mmp20 expression to
restore the correct substrate-to-enzyme ratio essential for healthy enamel.

Methods
Overexpression of Ambn and Mmp20 was achieved in separate mouse lines from transgenes (Tg). Ambn-Tg mice exhibited MIH-like phenotypes, and Mmp20-Tg mice displayed hypoplastic amelogenesis imperfecta. Crossbreeding of Ambn-Tg with Mmp20-Tg generated double-Tg mice overexpressing Ambn and Mmp20. Mandibular incisors of 7-week old mice (n=5/grp) were analyzed from the apical region to the incisal edge in sequential 1.0 mm volumes of interest (5 µm pixels) using isotropic micro-CT. Low, medium, and high thresholds were applied to quantify enamel volume and percent mineral volumes. Enamel mineral density was
calibrated to hydroxyapatite phantoms (.25 and .75 g/cm3). We compared the Ambn-Tg+Mmp20-Tg to the wildtype (WT) controls. We ran a single factor ANOVA test and a two-tailed T-test to determine if there is a significant difference between the groups.

Results
At eruption, the volumes of middle and high-density enamel in the Ambn and Mmp20 Tg’s demonstrated significant differences from the WT. Ambn-Tg had more middle density enamel volume (p=0.007). The Mmp20-Tg had significantly greater middle density (p=0.00001) and less higher density enamel (p=0.0000001) volumes compared to the WT, respectively. Ambn-Tg+Mmp20-Tg middle density and high-density enamel volumes were similar to the WT. Calculated enamel mineral densities were 38% (p=0.00001) and 8% (p=0.0005) lower in the Mmp20-Tg and Ambn-Tg groups compared to the WT, respectively. The Ambn-Tg+Mmp20-Tg
enamel mineral density was similar to the WT. Visual analysis indicated that the incisor enamel of Mmp20-Tg and Ambn-Tg groups was distributed in circular patches containing regions of lower and higher enamel density. A similar visual inspection indicated that the Ambn-Tg+Mmp20-Tg enamel was distributed like the WT.

Conclusion
These results show that enamel mineralization is highly sensitive to the balance of Ambn and Mmp20 expression. Imbalances can cause hypomineralized or hypoplastic enamel, while restoring the substrate-to-enzyme ratio can rescue normal enamel formation. This underscores the importance of maintaining balanced extracellular matrix protein and enzyme levels for healthy, functional enamel development.

Rebalancing the Substrate-Enzyme Ratio in Enamel Rescues Molar Incisor

Background
Despite advancements in genomic research, minoritized populations remain underrepresented, limiting the equitable distribution of benefits and contributing to ongoing health disparities. The Rare Genomes Project (RGP) at the Broad Institute of MIT and Harvard offers free whole genome sequencing to individuals in the U.S. with rare, suspected monogenic conditions who lack a genetic diagnosis. Between 2017 and 2021, enrollment was limited to English-proficient individuals. As a result, >90% of participants had some college education and >60% reported household incomes over $90,000, and no participants spoke a language other than English. Recognizing the lack of demographic diversity, RGP implemented a new diversity-focused study protocol in 2021 to address barriers to participation and increase access among historically underserved populations. 

Methods
To improve accessibility, RGP adopted strategies such as a simplified physician referral form, expanded language access (Spanish materials and interpreters), mobile phlebotomy, phone support, and extended staff hours. From 2021 to 2024, a total of 68 participants were enrolled who met at least one of six criteria for underrepresentation: non-white race, Hispanic/Latinx ethnicity, low income, limited English proficiency, rural residence, or lower educational attainment. Demographic and survey data were collected at baseline and sequencing was completed for all participants.

Results
Post-implementation, RGP saw an 8% decrease in the total proportion of white participants and a 5.5% increase in Hispanic participants. The proportion of non-English speakers increased by 3.3%. However, rural enrollment declined after 2022, despite offering mobile phlebotomy and flexible contact hours. Of 437 total RGP participants, 68 comprised the new underrepresented cohort. This group was more likely to have lower education and income and greater use of government insurance (62% vs. 36%, p<0.001). At baseline, the underrepresented cohort reported higher perceived importance of a genetic diagnosis and slightly higher stress levels. While both groups agreed a diagnosis would aid understanding and preparation, the underrepresented group prioritized its role in identifying better treatments. Challenges such as limited internet or transportation and provider trust remained barriers for rural and minoritized communities.

Conclusion
The revised RGP protocol showed some success in improving access and enrollment for underrepresented populations, especially through multilingual support and flexible engagement strategies. However, limited internet access and reduced rural participation highlighted remaining gaps. Patient-provider trust also emerged as a potential barrier. Future efforts will include participant interviews and follow-up surveys to better understand values and inform long-term engagement strategies. Continued enhancements and community-centered approaches are needed to ensure equitable access to genomic medicine.


The Impact of Enhanced Recruitment and Enrollment Support
on the Diversity of a Genomic Research Study

Background
In the last century, Influenza A viruses have caused the most global pandemics. The global panzootic of Highly Pathogenic Avian Influenza A (H5N1), its expanding mammalian host range, first mammalian-to-human transmissions and rising human infections are ominous. As a leading pandemic risk, early H5N1 detection is crucial to outbreak response but currently remains limited to the CDC and state public health laboratories. Here, we present a simple, rapid, adaptable, high throughput multiplex fragment analysis test for Influenza A (H5N1, H3N2, H1N1) subtyping and Influenza A/B, RSV and SARS-CoV-2 identification.

Methods
From the most up to date viral genomes we designed novel primers targeting conserved pathogenic regions across clinically relevant Influenza subtypes and generated unique gene-specific subtype signatures for H5N1 Influenza, endemic Influenza strains, RSV and SARS-CoV-2 via fragment analysis by capillary electrophoresis. Analytic sensitivity was determined by limit of detection analysis of Influenza A viral RNA in replicate reactions for each input amount of the viral RNA genome. Inter-/intra-run variability was assessed for all gene targets. Analytical specificity was established in evaluations of H5 and non-H5 human and avian Influenza A virus genomic RNA and by in silico analysis of potential primer cross-reactivity with other pathogens using BLAST® suite analysis.

Results
We generated unique fragment size-based signatures for each targeted gene. Targeted conserved regions in the HA gene are conserved in all newly sequenced U.S. Human H5N1 cases. The generated H5 amplicon had CT-values 2-12 lower than CDC and WHO H5 primers. No confounding false positives or over-lapping non-specific amplicons were identified. 100% H5N1 detection sensitivity was observed with a lower limit of detection of 2.5 copies/uL.

Conclusion
We have applied fragment analysis to Influenza A subtyping and respiratory virus detection for the first time. The multiplex target fragment analysis test described here has 100% analytical sensitivity and no false positives. This enables total subtyping capability of all human circulating Influenza A strains in one assay. This platform also proposes an epidemiological tool for detecting possible H5 Avian strain cross-over via dual detection of the H5 and M gene without the N1 amplicon. The use of multiple gene-specific targets for each strain increases test rigor, reproducibility and confidence in the accuracy of results.

Seasonal and Avian (H5N1) Influenza Subtyping by a Multiplex Fragment Analysis Test

Background: 
Vitiligo affects an estimated 0.5–2% of individuals worldwide and is characterized by cutaneous depigmentation. Recent studies suggest that autoimmune mechanisms in vitiligo extend beyond the skin, including ocular inflammation and auditory dysfunction. Notably, a nationwide Korean cohort of over 140,000 vitiligo patients found significantly elevated risks of dry eye, cataracts, and glaucoma compared with matched controls. However, comprehensive data from diverse U.S. adult populations remain scarce.

Objective: 
To determine whether adults with vitiligo have increased odds of developing ocular and auditory comorbidities compared with matched controls.

Methods: 
We conducted a retrospective, matched case–control study using de‑identified EHR data from the NIH All of Us Research Program. We identified cases based on the presence of ≥2 EHR visits with ICD‑10 L80 codes. Controls were matched 4:1 on age, sex, race/ethnicity, and duration of follow-up. We identified ocular outcomes (dry eye syndrome, uveitis, cataracts, glaucoma, retinal diseases, visual impairment) and auditory outcomes (sensorineural and sudden hearing loss, Ménière’s disease, benign paroxysmal positional vertigo, vestibular neuronitis) via validated ICD‑10 codes. Multivariate logistic regression estimated adjusted odds ratios (aORs) with 95% confidence intervals (CIs). Covariates included body mass index, Charlson Comorbidity Index (CCI), cumulative systemic steroid exposure, hypertension, diabetes, dyslipidemia, and chronic kidney disease. CCI is a validated score that assigns weighted points to chronic conditions based on ICD-10 codes in order to quantify baseline comorbidity burden.

Results: 
A total of 842 vitiligo cases and 3,368 matched controls were identified. Patients with vitiligo had lower BMI (28.91 vs 29.72 kg/m²; p=0.005), higher rates of long‑term steroid exposure (78.3% vs 56.6%; p<0.001), and higher proportions of CCI>1 (90.1% vs. 78.9%, p<0.001). Patiens with vitiligo patients exhibited higher unadjusted prevalence of any ocular comorbidity (43.2% vs 31.5%) and any auditory comorbidity (20.8% vs 15.9%). In fully adjusted models, vitiligo was associated with dry eye syndrome (aOR 1.39; 95% CI 1.13–1.71; p=0.002), anterior uveitis (aOR 3.00; 95% CI 1.18–7.64; p=0.021), cataracts (aOR 1.22; 95% CI 1.02–1.46; p=0.030), overall retinal disease (aOR 1.27; 95% CI 1.01–1.60; p=0.042), and other retinopathies (aOR 1.39; 95% CI 1.04–1.87; p=0.026). No adjusted associations persisted for any auditory comorbidity (aOR 1.04; 95% CI 0.85–1.27; p=0.703).

Conclusion: 
This large, multiethnic U.S. cohort demonstrates that vitiligo is independently associated with a spectrum of ocular comorbidities but not with auditory disorders. These findings underscore the need for routine ophthalmologic screening in patients with vitiligo and suggest that interdisciplinary care models may optimize long‑term visual health in this population.


Ocular and Auditory Comorbidities in Adults with Vitiligo: A Retrospective Case–Control Study

Background 

Shoulder injuries are common in adolescent athletes, particularly in overhead and contact sports. However, comparative data on sport-specific risk profiles remain limited. This study aimed to create risk profiles using strength and range-of-motion (ROM) of the shoulder across high school athletes in baseball, softball, female basketball, female wrestling, and male wrestling.

Methods 

We conducted a cross-sectional analysis of 247 high school athletes (159 male, 88 female) from three Utah schools between November 2023 and April 2025. Bilateral shoulder ROM and isometric strength were assessed using goniometry and handheld dynamometry. Primary outcomes included ER:IR strength ratio, internal rotation (IR) asymmetry, and total arc of motion (TROM) asymmetry. Athletes were classified as “at risk” if they met any of the following thresholds: ER:IR ratio < 0.75, IR deficit > 20°, or TROM asymmetry > 10°. Chi-square tests were used for between-group comparisons. 

Results 

Of the 247 athletes, 57 played baseball, 38 softball, 26 girls’ basketball, 24 girls’ wrestling, and 102 male wrestling. The most prevalent risk factor was ER:IR strength ratio < 0.75, observed in 45% of male wrestlers, 42% of female wrestlers, and 27% of girls’ basketball players. TROM asymmetry > 10° was most common in softball athletes (63%), followed by girls’ basketball (35%) and girls’ wrestling (33%). IR deficits > 20° were uncommon (≤12%) across all groups. Overall, 66% of male wrestlers and 63% of female wrestlers met at least one risk criterion. Significant between-sport differences were observed for ER:IR ratio (p < 0.001) and TROM asymmetry (p = 0.011), but not for IR deficit (p = 0.83). 

Conclusion 

Distinct sport-specific shoulder risk patterns were identified among adolescent athletes. ER:IR strength imbalance was most common in wrestlers, TROM asymmetry in softball and basketball players, and IR deficits were infrequent. These findings support the need for targeted screening and individualized preventive strategies to address modifiable shoulder injury risk in youth athletes. 



Sport-Specific Shoulder Risk Profiles in Adolescent Athletes: A Cross-Sectional Study

Background: Retinal detachment (RD) is a vision-threatening emergency requiring prompt surgical intervention with pars plana vitrectomy (PPV) (first-line), and/or scleral buckling (SB). PPV removes vitreous and repairs retinal detachment internally, while SB indents the sclera externally to reattach the retina. Though primary surgery is often successful, reoperations for detachments increase risks of permanent vision loss. This study identifies key anatomical predictors for indications of adjunctive SB to optimize single-surgery success rate (SSSR).

Methods: This retrospective cohort study (2010–2023) analyzed 3,528 RD repairs, including PPV alone (n = 2,811), SB alone (n = 163), and PPV+SB (n = 554). The primary outcome was SSSR at 3 months. Anatomical predictors included detachment extent (quadrants: Q1=superior nasal, Q2=superior temporal, Q3=inferior temporal, Q4=inferior nasal), macular status, lens status, gender, and prior ocular history. Chi-square tests compared outcomes; subgroup analyses excluded SB-only cases (secondary analysis).

Results:
Overall SSSR was 86.4% for PPV alone versus 89.5% for PPV+SB (p= 0.044). SB-only achieved 89.6% SSSR (p=0.321 vs PPV+SB).

Macular status: PPV+SB significantly outperformed PPV alone in macula-on detachments (93.5% vs 87.2%, p=0.005) but not macula-off cases (86.2% vs 85.9%, p=0.886).

Lens status: Phakic eyes benefited from PPV+SB (89.7% vs 85.3%, p=0.036), while pseudophakic eyes showed no difference (88.9% vs 87.3%, p=0.489).

Regional patterns: PPV+SB superiority was present when: Q3/Q4 were involved (Q3: 89.8% vs 84.9%, p=0.014; Q4: 89.7% vs 84.0%, p=0.013) and when Q1/Q2 were spared (Q1 absent: 90.6% vs 86.0%, p=0.025; Q2 absent: 92.5% vs 86.2%, p=0.009).

Panretinal detachments showed no significant PPV+SB benefit (84.9% vs 78.8%, p=0.249), whereas limited detachments trended positively (90.5% vs 87.2%, *p* = 0.052).

Demographics: Females had higher SSSR with PPV+SB (90.7% vs 84.5%, p=0.020), unlike males (88.7% vs 87.5%, p=0.533).

Conclusion: Adjunctive SB is associated with improved SSSR in specific anatomical contexts: macula-on detachments, phakic eyes, female patients, and detachments involving inferior quadrants (Q3/Q4) or sparing superior quadrants (Q1/Q2). These findings support selective SB use in RD repair, tailored to patient-specific anatomical risk profiles to avoid reoperations and improve visual outcomes.

Adjunctive Scleral Buckling with Pars Plana Vitrectomy for Retinal Detachment: Impact on Single-Surgery Success Rates by Anatomical Predictors

Chronic Shoulder Pain as a Herald of Advanced Small Cell Neuroendocrine Carcinoma
Background:
Small cell neuroendocrine carcinoma (SCNC) most commonly originates in the lung but can come from many organs. It typically presents with pulmonary symptoms such as cough, shortness of breath, or hemoptysis. We present a case of extensive-stage SCNC initially misattributed to benign shoulder pathology, ultimately requiring emergent chemotherapy intervention. Musculoskeletal pain as the sole initial manifestation presented a rare and challenging diagnostic scenario.
Case presentation:
A 51-year-old male with a 15-pack-year smoking history and family history of malignancies (mother with rectal cancer at age 48, and brother with head and neck cancer) presented with 2 months of right scapular and bilateral arm pain, initially managed with NSAIDs and chiropractic care. Symptoms progressed to right arm weakness, nocturnal pain, and 20-pound unintentional weight loss. Physical examination revealed right arm weakness (4/5) and numbness with otherwise stable vital signs apart from hypertension.
Initial CT abdomen obtained revealed extensive hepatic lesions suspicious for metastatic disease. Subsequent MRI cervical spine demonstrated osseous lesion in clivus and T1 vertebral body. Further evaluation with PET-CT showed FDG-avid metastatic involvement of the liver and numerous bones in the body, and significant neck and mediastinal lymphadenopathy. Notably, bone lesions were CT-occult, undetectable on prior imaging. Given the concerning imaging findings and rapid clinical progression, emergent chemotherapy was initiated due to the high risk of impending organ dysfunction from extensive tumor burden. Liver biopsy results demonstrate SCNC, supported by a high Ki-67 (70%) indicating rapid proliferation, positive synaptophysin confirming neuroendocrine differentiation, and elevated serum chromogranin A, further consistent with small cell neuroendocrine carcinoma. The patient demonstrated good initial tolerance to carboplatin/etoposide therapy with a plan to add immunotherapy after the first cycle.
Discussion:
This case highlights SCNC's potential to present atypically as isolated musculoskeletal pain, demonstrating how CT imaging may underestimate metastatic burden while PET-CT reveals more extensive disease. The diagnostic challenge is particularly acute in high-risk patients with significant smoking histories and familial cancer predisposition. These findings underscore the importance of maintaining clinical suspicion for malignancy even in the absence of classic symptoms, as earlier recognition can significantly impact patient outcomes.

Chronic Shoulder Pain as a Herald of Advanced Small Cell Neuroendocrine Carcinoma

Background
Social determinants of health (SDOH), particularly food insecurity and related socioeconomic factors, are increasingly recognized as critical influences on health outcomes. Extensive evidence highlights their profound role in shaping health disparities across various populations. This study examines associations between food security-related SDOH and cancer mortality and prevalence in the Kansas City Metropolitan Area. 

Methods
Using publicly available census tract-level data from the CDC PLACES, U.S. Census Bureau, and Neighborhood Atlas for the Kansas City Metropolitan Area (Missouri & Kansas) in 2023, linear regressions were performed to assess the relationships between cancer deaths and prevalence and key SDOH variables: total seniors (TractSeniors), housing units receiving SNAP (TractSNAP), population beyond 1/2 mile from a supermarket (lapophalf), and poverty rate (PovertyRate). Multicollinearity was assessed using VIF values; additionally, bivariate Pearson correlations were also performed. The analysis included 131 observations.

Results
Analysis revealed distinct predictors for cancer deaths versus prevalence. For cancer deaths, the model explained 57.8% of the variance, showing significant positive associations with higher proportions of seniors ( β=0.0198, p<0.001), SNAP recipients ( β=0.0211, p=0.001), and limited food access ( β=0.0016, p=0.001). PovertyRate was negatively associated with cancer deaths ( β=−0.1883, p=0.011). Excluding senior population intensified the impact of SNAP recipients ( β=0.0319, p<0.001), limited food access ( β=0.0027, p<0.001), and PovertyRate ( β=−0.3627, p<0.001) on cancer deaths. For crude cancer prevalence, the model explained 18.2% of the variance, with PovertyRate as the sole significant negative predictor (β=−0.0527, p<0.001). Bivariate correlations supported these findings.

Conclusion
Cancer deaths are strongly predicted by age, SNAP participation, and limited food access, underscoring their prominent and independent roles in cancer mortality. In stark contrast, crude cancer prevalence is less directly explained by these variables. The consistent negative association between poverty rate and both cancer deaths and prevalence is a critical and counter-intuitive finding that likely points to significant under-diagnosis or under-reporting in impoverished areas, potentially due to reduced healthcare access, rather than poverty itself being protective. These distinct findings underscore that different underlying drivers influence cancer mortality versus prevalence. Ultimately, this research underscores the profound impact of social determinants on health disparities in cancer outcomes, pinpointing specific census tracts within the Kansas City metropolitan area for targeted public health interventions and policy reform to address the true burden of cancer in vulnerable communities.


Unpacking the Plate: Food Insecurity as a Driver of Cancer Mortality in the Kansas City Urban Core

IL-21 and IL-21R interaction plays a key role in mounting humoral and cell-mediated immune response to adenoviral and adeno-associated viral vectors in lung

Authors: Mina R Hanna,1,2 Robert Clark,1 Alexander J Martin,1 Dong Lin,1 Nnamdi L Onyene,1 Xin Wang,1 and Jay Kolls1,2

1Center for Translational Research in Infection and Inflammation and 2Department of Pediatrics, Tulane School of Medicine, New Orleans, LA, USA

Purpose of Study: Although cystic fibrosis (CF) was discovered ~ a century ago, it remains without an available cure for all mutations. 8-10% of CF patients remain without therapy options despite advancements in targeting misfolded CFTR. Gene therapy is an active area of investigation in hopes of tackling this unmet need; however, the need for re-dosing remains a challenge for any means of viral vector delivery. Thus, mitigating the immune response may be the key to overcoming this challenge. Given the central role of IL-21R in regulating T and B cell immune response, we hypothesized that targeting IL-21R may attenuate B and T cell responses and enhance secondary transgene expression. 

Methods: Naïve or IL21R-/- mice received 4x1010 viral particles and 1x1011 viral particles intratracheally of an Ad5 vector for first and repeat doses, respectively, at days 0 and ~27. Serum anti-vector antibodies were assayed 2 weeks post-dosing. All mice were euthanized 3 days following 2nd viral dose. Lung tissue was harvested for analysis of B cells and CD8 tissue resident memory (TRM) cells as well as secondary transgene expression. Similarly, naïve or IL21R-/- mice received 1x1011 vg of adeno-associated virus 6.2 (AAV6.2) by endotracheal intubation and monitored with In Vivo imaging system (IVIS) to assess for transgene expression. 

Summary of Results: At 2 weeks, serum anti-AdV IgG (P<0.05) and IgG1 (P<0.01) were significantly decreased in the IL21R-/- mice by 2-way ANOVA. Flow cytometry analysis showed a significant reduction of CD8 TRM cells in IL-21R-/- mice (P<0.01) by nonparametric T-test and a reduction in CD19 B cells (P<0.05) by 2-way ANOVA. Moreover, secondary transgene expression was significantly enhanced in IL-21R-/- mice compared to B6 controls (P<0.01). Similarly, in the context of AAV, the generation of anti-AAV IgG requires IL-21R signaling (P<0.0001). IVIS also showed significantly enhanced secondary transgene expression of AAV6.2 in IL-21R-/- (P<0.0001). 

Conclusion: Our results consistently showed how IL-21R and IL-21 play an important role in the development of humoral and CD8 TRM-mediated response to adenoviral and AAV vectors. This suggests how targeting IL-21R signaling may be the key to overcoming challenges for gene therapy using viral vectors as a means of delivery. Future studies will investigate the role of IL-21R in adult mice using antibody blockade. 


IL-21 and IL-21R interaction plays a key role in mounting humoral and cell-mediated immune responses to adenoviral and adeno-associated viral vectors in the lung

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