CD70 is a therapeutic target upregulated in EMT-associated EGFR tyrosine kinase inhibitor resistance.

Effective therapeutic strategies are needed for non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations that acquire resistance to EGFR tyrosine kinase inhibitors (TKIs) mediated by epithelial-to-mesenchymal transition (EMT). We investigate cell surface proteins that could be targeted by antibody-based or adoptive cell therapy approaches and identify CD70 as being highly upregulated in EMT-associated resistance. Moreover, CD70 upregulation is an early event in the evolution of resistance and occurs in drug-tolerant persister cells (DTPCs). CD70 promotes cell survival and invasiveness, and stimulation of CD70 triggers signal transduction pathways known to be re-activated with acquired TKI resistance. Anti-CD70 antibody drug conjugates (ADCs) and CD70-targeting chimeric antigen receptor (CAR) T cell and CAR NK cells show potent activity against EGFR TKI-resistant cells and DTPCs. These results identify CD70 as a therapeutic target for EGFR mutant tumors with acquired EGFR TKI resistance that merits clinical investigation.

Optimizing Cognitive Assessment Outcome Measures for Alzheimer's Disease by Matching Wordlist Memory Test Features to Scoring Methodology.

Cognitive assessment with wordlist memory tests is a cost-effective and non-invasive method of identifying cognitive changes due to Alzheimer's disease and measuring clinical outcomes. However, with a rising need for more precise and granular measures of cognitive changes, especially in earlier or preclinical stages of Alzheimer's disease, traditional scoring methods have failed to provide adequate accuracy and information. Well-validated and widely adopted wordlist memory tests vary in many ways, including list length, number of learning trials, order of word presentation across trials, and inclusion of semantic categories, and these differences meaningfully impact cognition. While many simple scoring methods fail to account for the information that these features provide, extensive effort has been made to develop scoring methodologies, including the use of latent models that enable capture of this information for preclinical differentiation and prediction of cognitive changes. In this perspective article, we discuss prominent wordlist memory tests in use, their features, how different scoring methods fail or successfully capture the information these features provide, and recommendations for emerging cognitive models that optimally account for wordlist memory test features. Matching the use of such scoring methods to wordlist memory tests with appropriate features is key to obtaining precise measurement of subtle cognitive changes.

Prevalence and Longevity of SARS-CoV-2 Antibodies Among Health Care Workers.

BACKGROUND: Understanding severe acute respiratory syndrome coronavirus 2 antibody prevalence in a spectrum of health care workers (HCWs) may provide benchmarks of susceptibility, help us understand risk stratification, and support enactment of better health policies and procedures. METHODS: Blood serum was sampled at enrollment and 8-week follow-up from HCWs (n = 3458) and from community first responders (n = 226) for immunoglobulin G (IgG) analyses. Demographics, job duties, location, and coronavirus disease 2019-related information were collected. RESULTS: The observed IgG antibody prevalence was 0.93% and 2.58% at enrollment (May/June) and 8-week follow-up (July/August), respectively, for HCWs, and 5.31% and 4.35% for first responders. For HCWs, significant differences (P < .05) between negative and positive at initial assessment were found for age, race, fever, and loss of smell, and at 8-week follow-up for age, race, and all symptoms. Antibody positivity persisted at least 8 weeks in all positive HCWs. CONCLUSIONS: We found considerably lower antibody prevalence among HCWs compared with other published studies. While rigorous safety process measures instituted in our workplace and heightened awareness at and outside of the workplace among our HCWs may have contributed to our findings, the significant discrepancy from our community prevalence warrants further studies on other contributing factors.

SARS-CoV-2 antibody prevalence in health care workers: Preliminary report of a single center study.

Serological surveys have been conducted to establish prevalence for COVID-19 antibodies in various cohorts and communities, reporting a wide range of outcomes. The prevalence of such antibodies among healthcare workers, presumed at higher risk for infection, has been increasingly investigated, more studies are needed to better understand the risks and infection transmission in different healthcare settings. The present study reports on initial sero-surveillance conducted on healthcare workers at a regional hospital system in Orange County, California, during May and June, 2020. Study subjects were recruited from the entire hospital employee workforce and the independent medical staff. Data were collected for job duties and locations, COVID-19 symptoms, a PCR test history, travel record since January 2020, and existence of household contacts with COVID-19. A blood sample was collected from each subject for serum analysis for IgG antibodies to SARS-CoV-2. Of 2,992 tested individuals, a total 2,924 with complete data were included in the analysis. Observed prevalence of 1.06% (31 antibody positive cases), adjusted prevalence of 1.13% for test sensitivity and specificity were identified. Significant group differences between positive vs. negative were observed for age (z = 2.65, p = .008), race (p = .037), presence of fever (p < .001), and loss of smell (p < .001), but not for occupations (p = .710). Possible explanation for this low prevalence includes a relatively low local geographic community prevalence (~4.4%) at the time of testing, the hospital's timely procurement of personal protective equipment, rigorous employee education, patient triage, and treatment protocol development and implementation. In addition, cross-reactive adaptive T cell mediated immunity, as recently described, may possibly play a greater role in healthcare workers than in the general population.

Use of virtual games for interactive learning in a pharmacy curriculum.

BACKGROUND AND PURPOSE: To evaluate student pharmacists' attitudes and satisfaction toward playing educational virtual games in the classroom. EDUCATIONAL ACTIVITY AND SETTING: Virtual games were played in the classroom setting. First year student pharmacists participated in two Mimycx quests in the Healthcare Communication and the Psychiatry/Neurology courses. Students were randomly assigned into teams and worked together to complete the assigned quest games. Completion of the pre- and post-quest questionnaires via Qualtrics was voluntary. FINDINGS: A total of 79 student pharmacists played the Mimycx quests. Only 66 students completed both pre- and post-quest questionnaires. Students indicated their familiarity with game concepts related to the virtual environment and avatars used in the study. The change in their attitudes and satisfaction about the Mimycx virtual learning experience was significant between the two learning time points. SUMMARY: The use of virtual gaming technology could enhance student pharmacists' learning and engagement in the classroom. Students benefitted from increased familiarity with virtual, educational gaming concepts in their experiences with Mimycx, although no statistically significant differences were found regarding their attitudes toward communication and teamwork.

First-in-human, phase I/IIa dose-escalation and safety study of balugrastim in breast cancer patients receiving myelosuppressive chemotherapy.

PURPOSE: To evaluate safety of balugrastim, a recombinant human serum albumin and granulocyte colony-stimulating factor (G-CSF), administered over a range of therapeutic doses in women with breast cancer receiving doxorubicin plus docetaxel chemotherapy. METHODS: The phase I, sequential dose-escalation first segment compared subcutaneous balugrastim 50, 150, 300, and 450 µg/kg during chemotherapy cycles 0-2. The randomized (2:2:1), open-label, phase IIa second segment compared balugrastim 300 or 450 µg/kg with pegfilgrastim 6 mg during chemotherapy cycles 1 and 2. RESULTS: In the phase I segment, balugrastim was escalated to 450 µg/kg in 13 patients without dose-limiting toxicity. Three (9.7 %) of the 31 adverse events (AEs) reported in nine patients were grade 3 (agranulocytosis, vomiting, hypertension); none was grade 4. In the open-label phase IIa segment (N = 51), the majority of the 64 AEs reported in 31 (75.6 %) balugrastim-treated patients were grade 1 (59.4 %), with 39.1 % grade 2, 1.6 % grade 3 (one AE of vomiting), and none grade 4. Of the 16 AEs reported in seven (70.0 %) pegfilgrastim-treated patients, 87.5 % were grade 1, 6.3 % were grade 2, 6.3 % were grade 3 (one AE of thrombocytopenia), and none were grade 4. Overall, there were six bone pain AEs reported, one in the balugrastim 300 µg/kg group and five in the balugrastim 450 µg/kg group. No AEs in either study necessitated treatment interruption/discontinuation. The incidence and duration of grade 3-4 neutropenia were similar between balugrastim- and pegfilgrastim-treated patients. CONCLUSIONS: Balugrastim was well tolerated in this small population of breast cancer patients.

Development of a long-acting insulin analog using albumin fusion technology.

The primary therapeutic goal for the treatment of diabetes is maintenance of a long-term, near-normoglycemic condition and prevention of the onset or progression of the complications associated with the disease. Although several analogs of human insulin have been developed, the currently prescribed long-acting insulin analogs do not provide a stable basal glycemia for more than a few hours. Here, we report the development of Albulin, a long-acting insulin analog obtained by direct gene fusion of a single-chain human insulin to human serum albumin. Albulin showed an elimination t(1/2) of approximately 7 h in normoglycemic mice. In vitro pharmacodynamic profiles for Albulin characterized by receptor binding, inhibition of gluconeogenesis, induction of glucose uptake, and global regulation of gene expression in relevant cell types showed that Albulin produced similar activity profiles compared with that of recombinant human insulin. A single Albulin administration in vivo normalized blood glucose level in diabetic mice in a relatively peakless and sustained (24-h) fashion. A further reduction in glucose levels was achieved by administering a recombinant human insulin a few hours after Albulin injection in mice, indicating the potential for Albulin therapy in combination with available fast-acting insulin derivatives. In summary, Albulin displays characteristics of a potent long-acting insulin analog that can be evaluated for use as a novel insulin therapy for patients with insulin-dependent diabetes.

Syntaxin-6 SNARE involvement in secretory and endocytic pathways of cultured pancreatic beta-cells.

In pancreatic beta-cells, the syntaxin 6 (Syn6) soluble N-ethylmaleimide-sensitive factor attachment protein receptor is distributed in the trans-Golgi network (TGN) (with spillover into immature secretory granules) and endosomes. A possible Syn6 requirement has been suggested in secretory granule biogenesis, but the role of Syn6 in live regulated secretory cells remains unexplored. We have created an ecdysone-inducible gene expression system in the INS-1 beta-cell line and find that induced expression of a membrane-anchorless, cytosolic Syn6 (called Syn6t), but not full-length Syn6, causes a prominent defect in endosomal delivery to lysosomes, and the TGN, in these cells. The defect occurs downstream of the endosomal branchpoint involved in transferrin recycling, and upstream of the steady-state distribution of mannose 6-phosphate receptors. By contrast, neither acquisition of stimulus competence nor the ultimate size of beta-granules is affected. Biosynthetic effects of dominant-interfering Syn6 seem limited to slowed intragranular processing to insulin (achieving normal levels within 2 h) and minor perturbation of sorting of newly synthesized lysosomal proenzymes. We conclude that expression of the Syn6t mutant slows a rate-limiting step in endosomal maturation but provides only modest and potentially indirect interference with regulated and constitutive secretory pathways, and in TGN sorting of lysosomal enzymes.

Three-dimensional structure of the amino-terminal domain of syntaxin 6, a SNAP-25 C homolog.

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins are required for intracellular membrane fusion, and are differentially localized throughout the cell. SNAREs on vesicle and target membranes contain "SNARE motifs" which interact to form a four-helix bundle that contributes to the fusion of two membranes. SNARE motif sequences fall into four classes, homologous to the neuronal proteins syntaxin 1a, VAMP 2, and the N- and C-terminal SNARE motifs of SNAP-25 (S25N and S25C), and it is thought that one member from each class interacts to form a SNARE complex. Many SNAREs also feature N-terminal domains believed to function in regulating SNARE complex assembly or other aspects of vesicle transport. Syntaxin 6 is a SNARE found primarily in endosomal transport vesicles and whose SNARE motif shows significant homology to both syntaxin 1a and S25C. The crystal structure of the syntaxin 6 N-terminal domain reveals strong structural similarity with the N-terminal domains of syntaxin family members syntaxin 1a, Sso1p, and Vam3p, despite a very low level of sequence similarity. The syntaxin 6 SNARE motif can substitute for S25C in in vitro binding experiments, supporting the classification of syntaxin 6 as an S25C family member. Secondary structure prediction of SNARE proteins shows that the N-terminal domains of many syntaxin, S25N, and S25C family members are likely to be similar to one another, but are distinct from those of VAMP family members, indicating that syntaxin, S25N, and S25C SNAREs may have shared a common ancestor.