Design of a mucin-selective protease for targeted degradation of cancer-associated mucins.

Targeted protein degradation is an emerging strategy for the elimination of classically undruggable proteins. Here, to expand the landscape of targetable substrates, we designed degraders that achieve substrate selectivity via recognition of a discrete peptide and glycan motif and achieve cell-type selectivity via antigen-driven cell-surface binding. We applied this approach to mucins, O-glycosylated proteins that drive cancer progression through biophysical and immunological mechanisms. Engineering of a bacterial mucin-selective protease yielded a variant for fusion to a cancer antigen-binding nanobody. The resulting conjugate selectively degraded mucins on cancer cells, promoted cell death in culture models of mucin-driven growth and survival, and reduced tumor growth in mouse models of breast cancer progression. This work establishes a blueprint for the development of biologics that degrade specific protein glycoforms on target cells.

The Interprofessional Teaching Observation Program: A Faculty Development Workshop on Peer Feedback of Interprofessional Teaching.

Introduction: Faculty development focused on interprofessional education (IPE) is essential to any IPE initiative aiming to produce a collaborative practice-ready workforce. Many faculty have not received IPE in their own training and struggle with interprofessional teaching. Methods: To train faculty to conduct a peer-teaching observation and provide feedback focused on interprofessional teaching, we created a 3-hour didactic and skills practice workshop. The didactic portion considered ways interprofessional teaching differed from uniprofessional teaching, discussed elements of effective feedback, and reviewed the critical steps of a peer-teaching observation. In the skills practice portion, participants watched videos of different teaching scenarios and role-played as a peer observer providing feedback to the instructor in the videos. Participants completed a pre/post self-assessment and workshop evaluation form. Results: Eighteen faculty from four professions (dentistry, medicine, nursing, and pharmacy) participated in the workshop from 2020 to 2021. On a 5-point scale (1 = poor, 5 = excellent), participants rated the overall workshop quality 4.9 and the likelihood of making a change in their teaching/professional practice 4.8. Workshop participants' self-reported ability to provide feedback to a peer on their interprofessional teaching improved after workshop participation (preworkshop M = 2.9, postworkshop M = 3.8, p < .01). Discussion: This IPE-focused faculty development workshop allows participants to practice skills and share their own interprofessional teaching insights and challenges. The workshop is adaptable for different professions and settings and for in-person or online implementation. It also can be integrated into an existing program or utilized as a stand-alone workshop.

TeleBBD: Telehealth management of bladder bowel dysfunction - A provider survey of benefits and limitations.

INTRODUCTION: Bladder bowel dysfunction (BBD), defined by the International Children's Continence Society (ICCS) as a spectrum of lower urinary tract and bowel symptoms, represents to up to 40% of pediatric urology consults. Management of BBD involves strict behavioral management with frequent follow ups by urology advanced practice providers (APPs). If left untreated, patients may develop secondary comorbidities that impact their renal and/or bladder function, bowel function, and psychosocial well-being. Previous studies have reported feasibility for virtual post-operative visits and prenatal consultations, however, telehealth management of BBD, or TeleBBD, has not yet been studied. The goal of this study is to survey APPs in pediatric urology to understand how TeleBBD compares to in-person visits, and identify benefits and limitations of TeleBBD. STUDY DESIGN: An online survey via Qualtrics was designed based on current practice guidelines for BBD management and telehealth considerations. Survey was distributed in September 2020 via the Pediatric Urology Nurses & Specialists listserv. Group qualitative coding was completed by the authors to generate themes that emerged from the results. RESULTS/DISCUSSION: A total of 53 APPs from across 21 states in the US completed the online survey, with 49 (92%) APPs reporting currently providing TeleBBD. Those who did not provide TeleBBD typically do not manage BBD patients. Summary Table shows the comparison of TeleBBD with in-person visits, with many elements of TeleBBD better or the same as in-person visits. APPs reported that TeleBBD has been most beneficial in increasing access and overall improvement in follow up which is significant for management of this chronic condition. Barriers include access to technology/internet and inability to perform full physical exams via televisit. Limitations of the study include lack of validated survey and small sample size. Overall increased access and improved patient adherence and resolution from BBD is significant for this patient population for prevention of secondary comorbidities. CONCLUSION: This is the first study to survey APPs across the US specifically exploring indication of TeleBBD as well as comparison of TeleBBD with in-person visits. The perception of improved access to care and less no-show rates is significant especially during the pandemic in providing continuity of care and prevent secondary comorbidities. Additionally, TeleBBD was felt to be just as effective as in-person visits for patient adherence to treatment, and other components of care, with the exception of ability to provide physical exams. Providers leveraged the benefits of TeleBBD and shared strategies for best practices.

Classification, structural biology, and applications of mucin domain-targeting proteases.

Epithelial surfaces throughout the body are coated by mucins, a class of proteins carrying domains characterized by a high density of O-glycosylated serine and threonine residues. The resulting mucosal layers form crucial host-microbe interfaces that prevent the translocation of microbes while also selecting for distinct bacteria via the presented glycan repertoire. The intricate interplay between mucus production and breakdown thus determines the composition of the microbiota maintained within these mucosal environments, which can have a large influence on the host during both homeostasis and disease. Most research to date on mucus breakdown has focused on glycosidases that trim glycan structures to release monosaccharides as a source of nutrients. More recent work has uncovered the existence of mucin-type O-glycosylation-dependent proteases that are secreted by pathogens, commensals, and mutualists to facilitate mucosal colonization and penetration. Additionally, immunoglobulin A (IgA) proteases promote bacterial colonization in the presence of neutralizing secretory IgA through selective cleavage of the heavily O-glycosylated hinge region. In this review, we summarize families of O-glycoproteases and IgA proteases, discuss known structural features, and review applications of these enzymes to glycobiology.

Teleprecepting: A timely approach to clinical education during COVID-19.

ABSTRACT: Clinical training is a key component of nurse practitioner (NP) education. The rapid shift to telehealth necessitated by the pandemic has also created a need for socially distanced education and precepting. This article presents teleprecepting as a modality for clinical opportunities and connecting students to previously limited experiences, such as training in small specialties, in rural areas, and with interdisciplinary teams. Precepting NP students using telehealth follows similar principles as in-person teaching, but some modifications and additional considerations are needed to transition to the virtual environment. At a time when many NPs have swiftly adapted to telehealth in practice, this article will offer a brief "how-to" for teleprecepting. Prior to COVID-19, teleprecepting was piloted with less than 2% of NP students in the school's pilot teleprecepting project. Seven months after the initial surge of cases and restrictions, 72% of students (n = 151) in the family nurse practitioner, psychiatric mental health nurse practitioner, and pediatric nurse practitioner specialties had transitioned to teleprecepting. This project was implemented rapidly during the pandemic, and thus, evaluations comparing competency outcomes and experiences of students and preceptors are still in process. Additionally, feasibility of this educational model may change as telehealth regulations continue to evolve. COVID-19 poses challenges for both patient care and clinical training of NP students across specialties. With some adaptation, clinical placements can be transitioned to the virtual environment of telehealth. Future studies should examine student competencies based on teleprecepting experiences and preceptor training to support teleprecepting roles.

Shared Medical Appointments: An Academic-Community Partnership to Improve Care Among Adults With Type 2 Diabetes in California Central Valley Region.

PURPOSE: The purpose of the study was to evaluate the effectiveness of ALDEA (Latinos con Diabetes en Acción), a Shared Medical Appointment (SMA) intervention, compared to usual primary care (UPC) for the treatment of adults with type 2 diabetes over a 6-month period. It was hypothesized that participants in the SMA will have greater reductions in A1C at 6 months post-intervention compared to the control group. METHODS: This study was a quasi-experimental design with a non-randomized matched control group that followed participants prospectively for 6 months. All adults living with type-2 diabetes receiving primary care at a 2 FQHC clinics were eligible for inclusion. Participants in the control group were matched retrospectively on baseline A1C and age. RESULTS: The reductions in A1C were greater in the ALDEA SMA intervention group relative to the UPC control group at 6 months in both of the FQHC centers and in the combined sample. CONCLUSIONS: This study demonstrated that patients in the ALDEA program had a significantly greater reduction in A1C at 6 months compared to the control group. Despite its limitations, the ALDEA SMA program was successful in empowering patients and improving glycemic control.

Development of a general defined medium for Pichia pastoris.

Pichia pastoris is widely used as a host for recombinant protein production. More than 500 proteins have been expressed in the organism at a variety of cultivation scales, from small shake flasks to large bioreactors. Large-scale fermentation strategies typically employ chemically defined growth medium because of its greater batch-to-batch consistency and in many cases, lower costs compared to complex medium. For biopharmaceuticals, defined growth medium may also simplify downstream purification and regulatory documentation. Standard formulations of defined media for P. pastoris are minimal ones that lack the metabolic intermediates provided by complex components such as peptone and yeast extract. As a result, growth rates and per-cell productivities are significantly lower than in complex medium. We have designed a rich defined medium (RDM) for Pichia pastoris by systematically evaluating nutrients of increasing complexity and identifying those that are most critical for growth. We have also employed transcriptomics to gain deeper insights into the underlying metabolic processes and inform our media design. We have demonstrated that using RDM for expression of three heterologous proteins yields titers comparable to, or higher than, those in standard complex medium. RDM improves productivity of P. pastoris fermentations and its development demonstrates the usefulness of transcriptomics to accelerate process development for new molecules.

Cover Image, Volume 115, Number 1, January 2018.

Cover Legend: The cover image, by Catherine B. Matthews et al., is based on the Article Development of a general defined medium for Pichia pastoris, DOI 10.1002/bit.26440.

Reexamining opportunities for therapeutic protein production in eukaryotic microorganisms.

Antibodies are an important class of therapeutics and are predominantly produced in Chinese Hamster Ovary (CHO) cell lines. While this manufacturing platform is sufficiently productive to supply patient populations of currently approved therapies, it is unclear whether or not the current CHO platform can address two significant areas of need: affordable access to biologics for patients around the globe and production of unprecedented quantities needed for very large populations of patients. Novel approaches to recombinant protein production for therapeutic biologic products may be needed, and might be enabled by non-mammalian expression systems and recent advances in bioengineering. Eukaryotic microorganisms such as fungi, microalgae, and protozoa offer the potential to produce high-quality antibodies in large quantities. In this review, we lay out the current understanding of a wide range of species and evaluate based on theoretical considerations which are best poised to deliver a step change in cost of manufacturing and volumetric productivity within the next decade.Related article: http://onlinelibrary.wiley.com/doi/10.1002/bit.26383/full.