Publications by Year: 2026

Protein misfolding in the brain is a key pathological hallmark of neurodegenerative diseases. Optical imaging of misfolded proteins in disease models is essential for elucidating etiology and early diagnosis. However, developing specific optical imaging probes for each misfolded protein is time-consuming and challenging, leaving many pathological targets without effective detection tools, especially for in vivo imaging. Here, we present a dual-mode chemiluminescence strategy that enables both generic and specific detection of misfolded proteins using a single probe platform. In the generic mode, we demonstrate that ADLumin-1, a chemiluminescent probe, enables highly sensitive detection of diverse misfolded proteins in vitro, achieving up to 128-fold higher signal enhancement than Thioflavin T, and allows noninvasive imaging in mice models of Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis. In the specific mode, ADLumin-1 combined with protein misfolding cyclic amplification allows femtomolar-level detection of α-synuclein in cerebrospinal fluid, while integration with a bio-orthogonal chemiluminescence resonance energy transfer technique enables in vivo discrimination of α-synuclein from Aβ. This dual-mode, modular approach offers a practical solution to the current probe limitations, with potential preclinical and clinical applications in neurodegenerative disorders.

Over 90% of pancreatic ductal adenocarcinoma (PDAC) patients involve KRAS mutations (KRASMUT), for which current treatment options are limited. Statins, commonly used to lower cholesterol, have demonstrated certain selective toxicity towards KRAS-transformed cells, prompting the question of whether statin-based conjugates could achieve selective uptake specifically in KRASMUT cells. To investigate this, we synthesized statin-dye conjugates by attaching a fluorescent dye (Cy5.5) to two statins: simvastatin and pravastatin, aiming to assess whether selective uptake indeed occurs. Our findings revealed that these conjugates exhibited markedly enhanced uptake in KRASMUT cells compared to KRAS wild-type (KRASWT) cells. We evaluated the uptake of these conjugates in both KRASMUT and KRASWT cells and examined their potential to selectively target KRASMUT pancreatic cancer cells (PCCs) using an engineered PDAC tumor model co-cultured with PCCs and cancer-associated fibroblasts (CAFs). Our findings indicate that KRASMUT cancer cells exhibited higher uptake of statin-Cy5.5 conjugates via enhanced macropinocytosis compared to KRASWT cancer cells and CAFs. We also found enhanced uptake of the statin-Cy5.5 conjugate in MCF10A cells with PTEN deficiency, a condition known to elevate macropinocytosis, compared to control MCF10A cells with wild-type PTEN. Notably, in the PCC and CAF co-culture model, the pravastatin-Cy5.5 conjugate selectively killed KRASMUT PCCs without affecting the KRASWT CAFs. These findings highlight the unique synergistic potential of statin-Cy5.5-distinct from either component alone-as targeted delivery vehicles for KRASMUT cancer therapy.

Biological effects induced by diverse types of ionizing radiation are known to show important variations. Nanodosimetry is suitable for studying the link between these variations and the patterns of radiation interactions within nanometer-scale volumes, using experimental techniques complemented by Monte Carlo track structure (MCTS) simulations. However, predicted nanodosimetric quantities differ among MCTS codes, primarily because each code employs distinct molecular-scale particle interaction models. This multi-code study examines these variations for low-energy electrons (20-10,000 eV), which play a critical role in energy deposition and biological effects by virtually all types of ionizing radiation. Specifically, the hypothesis tested in this work is that inter-code variability in nanodosimetry results is mainly caused by differences in assumptions regarding total interaction cross sections. Ionization cluster size distributions and derived nanodosimetric parameters were simulated with seven MCTS codes (PARTRAC, PHITS-TS, MCwater, PTra, and three Geant4-DNA options) in liquid water as a surrogate for biological tissue. Significant inter-code differences were observed, especially at the lowest energies. They were substantially reduced upon replacing the original cross sections in each code with a common, averaged dataset, created ad-hoc for this study and not based on theoretical assumptions. For example, for 50 eV electrons in 8 nm spheres, the variability in the predicted mean ionization numbers decreased from 23% to 5%, and in the probability of inducing two or more ionizations from 34% to 7% (relative standard deviations). This quantification demonstrates that total interaction cross sections are the primary source of uncertainty at low electron energies. A sensitivity test using DNA damage simulations with the PARTRAC code revealed that cross section variations notably affect biological outcome predictions. Replacing the code's original cross sections with the averaged ones increased the predicted double-strand break yield by up to 15%. These findings underscore the urgent need for improved characterization of low-energy electron interaction cross sections to reduce uncertainties in MCTS simulations and enhance mechanistic understanding of radiation-induced biological effects.

IMPORTANCE: The clinical benefit of opportunistic genomic screening for familial hypercholesterolemia (FH) has not been demonstrated in a randomized clinical trial (RCT).

OBJECTIVE: To evaluate the impact of returning clinically confirmed FH-associated genetic results on low-density lipoprotein cholesterol (LDL-C) levels.

DESIGN, SETTING, AND PARTICIPANTS: This RCT was performed within the Veterans Health Administration, a large national health care system, and linked to the Million Veteran Program (MVP), a research biobank. Participants were MVP enrollees suspected to have an FH-associated genetic variant, as identified in their research data. Recruitment occurred from February 27, 2020, to September 20, 2022, and 6-month follow-up was completed October 21, 2024.

INTERVENTIONS: Delivery of clinical genetic confirmation testing and telegenetic counseling at baseline (immediate results arm) vs after 6 months (delayed results arm).

MAIN OUTCOMES AND MEASURES: Change in LDL-C levels (primary outcome) and proportions with treatment intensification and achievement of LDL-C target levels at 6 months (secondary outcomes).

RESULTS: The trial randomized 112 participants across 28 US states (mean age, 65.9 [range, 36-91] years; 94 [83.9%] men). Baseline mean (SD) LDL-C level was 109.5 (55.5) mg/dL, and 86 participants (76.8%) were already receiving therapy to lower lipid levels. At 6 months, the between-arm difference in LDL-C level reduction was -10.5 (95% CI, -21.9 to 1.0) mg/dL (P = .07; Cohen d = 0.34). Bayesian analysis suggested a high probability of benefit but was exploratory. Treatment was intensified in 11 of 55 participants (20.0%) in the immediate results arm vs 5 of 57 (8.8%) in the delayed results arm (P = .09). Fifteen participants (27.3%) in the immediate results arm vs 14 (24.6%) in the delayed results arm (P = .74) achieved LDL-C target levels. Thirty of 49 participants (61.2%) in the immediate results arm who completed this information shared their genetic result with a total of 98 relatives.

CONCLUSIONS AND RELEVANCE: In this RCT, opportunistic genomic screening for FH plus telegenetic counseling did not result in a statistically significant improvement in LDL-C levels and clinical management; however, the findings suggest that there may be a small to moderate benefit favoring the immediate results arm. Further research should be conducted to confirm these findings, optimize implementation strategies, and assess the long-term effects on cardiovascular outcomes.

TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT04178122.

BACKGROUND: ETS2 expression in prostate cancer (PCa) shows variability, with overexpression and loss reported. ETS2 can be dysregulated through proteasomal degradation, but p53 missense mutants may stabilize ETS2 by preventing this process. TP53 mutations are more frequent in metastatic PCa, and their identification in primary tumors could serve as an early marker for patient stratification. In addition, TMPRSS2-ERG fusion often arises from an intrachromosomal deletion including ETS2 locus.

METHODS AND RESULTS: ETS2, p53 and ERG protein expression were assessed by immunohistochemistry in a surgically treated PCa patients' cohort (N = 199). ETS2 and TMPRRS2-ERG mRNA were evaluated by qPCR in an independent series (N = 80). Relationship between molecular markers, and with clinicopathological characteristics was analyzed. High ETS2 protein expression was observed in 34.7% of tumors. Aberrant p53 immunostaining was detected in 5.5% of cases, being associated with high ETS2 expression (p = 0.050). p53 positivity was related to GG5 (p = 0.007) and unifocality (p = 0.011). Shorter time to PSA recurrence was observed in p53-positive patients (p = 0.002). Cases with high ETS2 expression but negative p53 staining had the longest time to recurrence, whereas p53-positive cases, regardless of ETS2, had the shortest (p = 0.002). ERG protein expression -surrogate marker of TMPRSS2-ERG- was strongly associated with high ETS2 (p < 0.001). At mRNA level, low ETS2 expression correlated with TMPRSS2-ERG fusion (p = 0.018), and showed a trend toward association with intrachromosomal deletion (p = 0.113) and high-stage (p = 0.054).

CONCLUSIONS: In conclusion, p53 staining was consistently associated with aggressive PCa and may serve as a reliable prognostic marker. High ETS2 expression correlates with delayed PSA recurrence in p53-negative tumors.

OBJECTIVE: We assessed the association between timing of moderate-to-vigorous physical activity (MVPA) with incident type 2 diabetes (T2D) and glycemic measures.

RESEARCH DESIGN AND METHODS: Regression models were used to assess associations between accelerometer-derived MVPA timing and incident T2D in UK Biobank (UKB) (n = 84,528, prospective), prevalent diabetes, and glycemic measures in the National Health and Nutrition Examination Survey (NHANES) (n = 6,998, cross-sectional).

RESULTS: In UKB, with early morning (0500-0959) MVPA as reference and before adjustment for total MVPA, "variable-timing" MVPA was associated with the lowest incident T2D risk; while after adjustment, afternoon-evening MVPA (1500-2400) showed the lowest incident T2D risk. In NHANES, afternoon/early evening MVPA was weakly associated with more favorable glycemic measures and lower diabetes prevalence after adjustment for total MVPA.

CONCLUSIONS: When keeping total MVPA volume constant, clustering MVPA in the afternoon-evening was associated with the strongest reduction in incident T2D risk, fewer prevalent diabetes, and more favorable glycemic measures.

Dysregulated actions of the bone-derived phosphaturic hormone, fibroblast growth factor-23 (FGF23), underlie the pathophysiology of several diseases. FGF23 is synthesized primarily in osteocytes in response to various endogenous molecules; however, the mechanisms governing FGF23 production are incompletely understood. Glycerol-3-phosphate (G3P), a glycolytic by-product originating from the kidney, critically controls skeletal FGF23 synthesis via its conversion in bone to lysophosphatidic acid (LPA), which stimulates osteocyte FGF23 production. The bioactive vitamin D, 1,25-dihydroxyvitamin D (1,25D), also promotes FGF23 production in osteocytes. We herein demonstrated that LPA requires 1,25D action to raise FGF23 levels in mouse bone explants and mice. RNA sequencing of osteocyte-like Ocy454 cells identified differentially expressed genes (DEGs) uniquely induced by LPA/1,25D co-treatment. These unique DEGs were enriched for the ribosome biogenesis pathway. DEGs concurrently induced by individual LPA and 1,25D treatments were enriched for MAPK signaling, and inhibiting this pathway obliterated LPA/1,25D-induced FGF23 production. DEGs following LPA/1,25D co-treatment were enriched for the cytokine-cytokine receptor interaction pathway. Moreover, LPA/1,25D co-treatment, but not individual LPA and 1,25D treatments, rapidly induced the expression of Il12a, the gene encoding the pro-inflammatory cytokine interleukin-12 alpha-subunit, which responded solely to 1,25D at later times and required MAPK-ERK1/2 signaling. Inhibiting cytokine signaling or knocking down Il12a inhibited, while overexpressing Il12a enhanced LPA/1,25D-induced FGF23 production. However, challenging Ocy454 cells with recombinant bioactive interleukin-12 failed to enhance FGF23 production, suggesting that Il12a plays a noncanonical role. Our results reveal a mechanism of skeletal FGF23 synthesis involving synergistic actions of LPA and 1,25D, advancing our understanding of FGF23 regulation.

(1) Background: Wound healing is a highly coordinated process encompassing hemostasis, inflammation, angiogenesis, keratinocyte migration, collagen deposition, and extracellular matrix remodeling. Successful repair also requires adequate nutrient and oxygen delivery through a well-developed vascular supply. Disruption of these processes can occur through aberrations in diverse biological pathways, including extracellular matrix organization, cellular adhesions, angiogenesis, and immune regulation. (2) Methods: We reviewed mechanisms of impaired tissue repair in monogenic disorders by focusing on three categories-connective tissue, hematological/immunological, and aging-related disorders-to illustrate how single-gene defects disrupt inflammation, cellular proliferation, and matrix remodeling. Additionally, we reviewed various polygenic disorders-chronic kidney disease, diabetes mellitus, hypertension, and obesity-to contrast complex multifactorial pathologies with single-gene defects. (3) Results: This review establishes that genetic impediments, despite their distinct etiologies, monogenic and polygenic disorders share critical downstream failures in the wound healing cascade. While monogenic diseases illustrate direct causal links between specific protein deficits and repair failure, polygenic diseases demonstrate how multifactorial stressors overwhelm the body's regenerative capacity. (4) Conclusions: This review synthesizes current evidence on both monogenic diseases and polygenic contributions to impaired wound healing. These findings highlight that genetic susceptibility is a decisive factor in the ability to restore tissue homeostasis. This underscores the profound impact of genetic background on the efficacy of hemostasis, inflammation, and remodeling.

INTRODUCTION: Flexible endoscopy (FE), rigid endoscopy (RE), and open surgery (OS) are established treatments of Zenker diverticulum (ZD); however, comparative data on their clinical outcomes remain limited and inconsistent. We conducted the first multicenter matched cohort study comparing several procedural outcomes across FE, RE, and OS for ZD repair.

METHODS: The study population was matched for age, sex, American Society of Anesthesiologists score, and diverticulum. Size resulting in outcomes assessed included dysphagia scores, clinical and technical success, adverse events (AEs), procedure length, hospital length of stay, and need for further interventions. Both overall and pairwise comparisons were conducted. Subgroup analyses were performed to assess for comparative efficacy of FE procedure types, effect of previous intervention on procedure success, and the effect the year the procedure was performed on clinical success.

RESULTS: There were 202 eligible patients over the study period. Propensity score matching resulted in 125 patients (FE n = 44, RE n = 36, OS n = 45). FE demonstrated higher clinical success than RE (84.6% vs 55.6%, P = 0.032) and similar success to OS (88.9%, P = 0.712); OS showed higher success than RE ( P = 0.014). AE rates were lowest with FE (4.5%) compared with RE (8.3%) and OS (11.1%) although this was not statistically significant ( P = 0.212). FE had shorter median length of stay (0 days vs 1 day for RE and 3 days for OS, P < 0.001) and median procedure lengths (55 minutes vs 58 minutes for RE and 78 minutes for OS, P = 0.018).

DISCUSSION: In this matched cohort, FE has clinical success comparable with OS and superior to RE for ZD repair, with shorter hospital stays, reduced procedure lengths, and a trend toward fewer AEs. Our matched, multicenter design provides stronger comparative evidence than previous unmatched studies. These findings support FE as a first-line treatment of ZD repair, although prospective studies could further clarify long-term outcomes.