Triple-negative breast cancer (TNBC) is an aggressive subtype defined by lack of hormone receptor expression and non-amplified HER2. Adavosertib (AZD1775) is a potent, small-molecule, ATP-competitive inhibitor of the Wee1 kinase that potentiates the activity of many DNA-damaging chemotherapeutics and is currently in clinical development for multiple indications. The purpose of this study was to investigate the combination of AZD1775 and capecitabine/5FU in preclinical TNBC models. TNBC cell lines were treated with AZD1775 and 5FU and cellular proliferation was assessed in real-time using IncuCyte^® Live Cell Analysis. Apoptosis was assessed via the Caspase-Glo 3/7 assay system. Western blotting was used to assess changes in expression of downstream effectors. TNBC patient-derived xenograft (PDX) models were treated with AZD1775, capecitabine, or the combination and assessed for tumor growth inhibition. From the initial PDX screen, two of the four TNBC PDX models demonstrated a better response in the combination treatment than either of the single agents. As confirmation, two PDX models were expanded for statistical comparison. Both PDX models demonstrated a significant growth inhibition in the combination versus either of the single agents. (TNBC012, p < 0.05 combo vs. adavosertib or capecitabine, TNBC013, p < 0.01 combo vs. adavosertib or capecitabine.) An enhanced anti-proliferative effect was observed in the adavosertib/5FU combination treatment as measured by live cell analysis. An increase in apoptosis was observed in two of the four cell lines in the combination when compared to single-agent treatment. Treatment with adavosertib as a single agent resulted in a decrease in p-CDC2 in a dose-dependent manner that was also observed in the combination treatment. An increase in γH2AX in two of the four cell lines tested was also observed. No significant changes were observed in Bcl-xL following treatment in any of the cell lines. The combination of adavosertib and capecitabine/5FU demonstrated enhanced combination effects both in vitro and in vivo in preclinical models of TNBC. These results support the clinical investigation of this combination in patients with TNBC, including those with brain metastasis given the CNS penetration of both agents.

Enterovirus D68 (EV-D68) caused large biennial cyclical outbreaks of respiratory disease and cases of acute flaccid myelitis from 2014 to 2018 in the USA. An anticipated outbreak did not occur in 2020, likely due to non-pharmaceutical interventions targeting the COVID-19 pandemic. A large respiratory disease outbreak occurred again in 2022, but uncertainty remained regarding if circulation of EV-D68 would return to the pre-pandemic patterns. We conducted prospective active surveillance of clinical respiratory specimens from Colorado children for EV-D68 in 2023 and 2024. A subset of residual specimens positive for rhinovirus/enterovirus (RV/EV) were tested for EV-D68 via a validated in-house EV-D68 reverse transcription–PCR assay. During epi weeks 18–44 in 2023, 525 residual specimens positive for RV/EV all tested negative for EV-D68. In 2024, during epi weeks 18–44, 10 (1.8%) of the 546 RV/EV-positive specimens were EV-D68-positive. The EV-D68-positive cases were predominantly young children (median age 4.8 years) receiving treatment with asthma medications. Following the 2022 EV-D68 outbreak, an anticipated outbreak did not occur in 2023. While EV-D68 was detected in 2024, the number of cases was not as significant as in prior outbreak years. Continued surveillance for EV-D68 will be important to understand the future dynamics of EV-D68 circulation and prepare for future outbreaks.

Background: Enterovirus D68 (EV-D68) causes cyclical outbreaks worldwide. Though EV-D68 can rarely cause acute flaccid myelitis, it commonly causes respiratory disease. In particular, EV-D68 causes outbreaks of severe asthma-like respiratory disease, but few systems have the capability to detect EV-D68 cases. Methods: Active, prospective EV-D68 surveillance at Children’s Hospital Colorado has been performed since 2014, where a random subset of residual clinical respiratory specimens, primarily from inpatient settings, and positive for rhinovirus/enterovirus are tested for EV-D68 via reverse transcription-polymerase chain reaction. Data via chart review from this surveillance were analyzed from 2014 to 2025 to characterize the clinical characteristics of EV-D68 respiratory disease in children (<18 years). Results: From 2014 to 2025, EV-D68 was detected in 308 patients, of which 271/308 (88%%) were children with respiratory disease. Median age of children with respiratory disease was 6 [interquartile range (IQR): 3.1–10.4] years; 162/271 (59.8%) were males. A majority of children admitted required intensive care unit admission (206/258, 79.8%). Though only 174/271 (64.2%) children had an atopic history, a significant majority required bronchodilator therapy (249/271, 91.9%) and systemic steroids (229/271, 84.5%). Median intensive care unit and hospital length of stay were 2 (IQR: 1–3) and 3 (IQR: 2–5) days, respectively. Conclusions: Children with EV-D68 respiratory disease commonly presented with severe asthma-like respiratory disease, even in those without atopy. Despite the severity of disease, children quickly improved over short hospitalizations when managed with bronchodilators, steroids, and respiratory support. With improved diagnostics to detect and recognize the true burden of EV-D68, additional studies evaluating short- and long-term management of EV-D68 respiratory disease are needed.