The Eyewitness Community Survey: An Engaging Citizen Science Tool to Capture Reliable Data while Improving Community Participants' Environmental Health Knowledge and Attitudes.

Many youths and young adults have variable environmental health knowledge, limited understanding of their local environment's impact on their health, and poor environmentally friendly behaviors. We sought to develop and test a tool to reliably capture data, increase environmental health knowledge, and engage youths as citizen scientists to examine and take action on their community's challenges. The Eyewitness Community Survey (ECS) was developed through several iterations of co-design. Herein, we tested its performance. In Phase I, seven youths audited five 360° photographs. In Phase II, 27 participants works as pairs/trios and audited five locations, typically 7 days apart. Inter-rater and intra-rater reliability were determined. Changes in participants' knowledge, attitudes, behaviors, and self-efficacy were surveyed. Feedback was obtained via focus groups. Intra-rater reliability was in the substantial/near-perfect range, with Phase II having greater consistency. Inter-rater reliability was high, with 42% and 63% of Phase I and II Kappa, respectively, in the substantial/near-perfect range. Knowledge scores improved after making observations (p ≤ 0.032). Participants (85%) reported the tool to be easy/very easy to use, with 70% willing to use it again. Thus, the ECS is a mutually beneficial citizen science tool that rigorously captures environmental data and provides engaging experiential learning opportunities.

Environmental Health Knowledge Does Not Necessarily Translate to Action in Youth.

Environmental challenges pose serious health problems, especially for children, and lay public action is lacking. This study sought to characterize the relationship between environmental health knowledge and behavior in youth. A cross-sectional, descriptive survey with quantitative and qualitative questions was conducted. Open-ended questions were coded to generate themes/subthemes. Subscales' scores were presented as mean ± SD or median and interquartile range (IQR). T- and Mann-Whitney tests were used to compare groups, and correlations were used to evaluate covariation. A total of 452 children were surveyed. Youth verbalized concerns about their environments and their impact on health. Air pollution was the most concerning issue. Participants had moderate knowledge scores. Few described the three health domains; even fewer included environment. Behavior scores were low and weakly correlated with knowledge, but were moderately correlated with attitude and self-efficacy. Participation in environmental classes, activities, and clubs was associated with higher scores. We found variable environmental health knowledge, limited understanding of the local environment's impact on health, and a weak association between youth's knowledge and behavior. Focused formal and non-formal educational experiences were associated with improved scores, indicating the value of targeted youth educational programming to increase environmental health knowledge and action.

Discovery of mobocertinib, a potent, oral inhibitor of EGFR exon 20 insertion mutations in non-small cell lung cancer.

In the treatment of non-small cell lung cancer (NSCLC), patients harboring exon 20 insertion mutations in the epidermal growth factor receptor (EGFR) gene (EGFR) have few effective therapies because this subset of mutants is generally resistant to most currently approved EGFR inhibitors. This report describes the structure-guided design of a novel series of potent, irreversible inhibitors of EGFR exon 20 insertion mutations, including the V769_D770insASV and D770_N771insSVD mutants. Extensive structure-activity relationship (SAR) studies led to the discovery of mobocertinib (compound 21c), which inhibited growth of Ba/F3 cells expressing the ASV insertion with a half-maximal inhibitory concentration of 11 nM and with selectivity over wild-type EGFR. Daily oral administration of mobocertinib induced tumor regression in a Ba/F3 ASV xenograft mouse model at well-tolerated doses. Mobocertinib was approved in September 2021 for the treatment of adult patients with advanced NSCLC with EGFR exon 20 insertion mutations with progression on or after platinum-based chemotherapy.

Shared Purpose: Leveraging a Community-Academic Partnership to Increase Local Environmental Health Awareness via Community Science.

Environmental factors can lead to disease and health disparities when the places where people live, learn, work, play and pray are burdened by social inequities. Non-formal programs that explicitly connect local environmental exposures and human health could be of great value to communities at greatest risk. The purpose of this work was to co-create relevant and engaging education with youth and community stakeholders of all ages that more explicitly emphasizes the link between the local environment and community members' health through a hands-on community science experience. Our experiences helped strengthen our community-academic partnership and establish a route to create and tailor informal programming to meet local needs and engage people in community science with academic partners. We generated two distinctly different community science neighborhood audit tools designed to differently engage our community partners and inform community participants of their local environments and its role on their health. Through community meetings, we garnered critical insight from our stakeholders. While neither of the tools and accompanying data collected were deemed to be scientifically generalizable, our ongoing and future work has benefited from important lessons learned from their creation and sharing.

Targeting HER2 Exon 20 Insertion-Mutant Lung Adenocarcinoma with a Novel Tyrosine Kinase Inhibitor Mobocertinib.

No targeted treatments are currently approved for HER2 exon 20 insertion-mutant lung adenocarcinoma patients. Mobocertinib (TAK-788) is a potent irreversible tyrosine kinase inhibitor (TKI) designed to target human epidermal growth factor receptor 2 (HER2/ERBB2) exon 20 insertion mutations. However, the function of mobocertinib on HER2 exon 20 insertion-mutant lung cancer is still unclear. Here we conducted systematic characterization of preclinical models to understand the activity profile of mobocertinib against HER2 exon 20 insertions. In HER2 exon 20 insertion-mutant cell lines, the IC50 of mobocertinib was higher than poziotinib and comparable with or slightly lower than afatinib, neratinib, and pyrotinib. Mobocertinib had the lowest HER2 exon 20 insertion IC50/wild-type (WT) EGFR IC50 ratio, indicating that mobocertinib displayed the best selectivity profile in these models. Also, mobocertinib showed strong inhibitory activity in HER2 exon 20YVMA allograft and patient-derived xenograft models. In genetically engineered mouse models, HER2 exon 20G776>VC lung tumors exhibited a sustained complete response to mobocertinib, whereas HER2 exon 20YVMA tumors showed only partial and transient response. Combined treatment with a second antibody-drug conjugate (ADC) against HER2, ado-trastuzumab emtansine (T-DM1), synergized with mobocertinib in HER2 exon 20YVMA tumors. In addition to the tumor cell autonomous effect, sustained tumor growth control derived from M1 macrophage infiltration and CD4+ T-cell activation. These findings support the ongoing clinical development of mobocertinib (NCT02716116) and provide a rationale for future clinical evaluation of T-DM1 combinational therapy in HER2 exon 20YVMA insertion-mutant lung adenocarcinoma patients. SIGNIFICANCE: This study elucidates the potent inhibitory activity of mobocertinib against HER2 exon 20 insertion-mutant lung cancer and the synergic effect of combined mobocertinib and T-DM1, providing a strong rationale for clinical investigation.

African American Breastfeeding Peer Support: All Moms Empowered to Nurse.

Background: Although breastfeeding is optimal infant nutrition, disparities in breastfeeding persist in the African American population. AMEN (Avondale Moms Empowered to Nurse) launched a Peer-to-Peer support group to increase breastfeeding initiation and duration in an under-resourced African American urban community with low breastfeeding rates. Materials and Methods: A Community-Based Participatory Research (CBPR)-guided project was developed in partnership with a neighborhood church. Using modified Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) peer counseling materials, Avondale neighborhood breastfeeding moms were trained and designated Breastfeeding Champions. Community organizations and partnering agencies helped recruit local mothers. Support groups included childcare, transportation, refreshments, and incentives, plus stipends for Champions. A mixed-methods approach captured participation, feeding intention and practices, and program evaluation using electronic data capture. After adding another neighborhood with low breastfeeding rates, AMEN was modified to "All Moms Empowered to Nurse." Additional Champion moms were trained as Reaching Our Sisters Everywhere (ROSE) Community Transformers. During the COVID-19 pandemic, the group has met weekly by virtual platform. Results: Since May 2017, 67 AMEN support meetings have included 158 participants, with average attendance of 10 (range 5-19) per meeting. In addition to 8 Champions, 110 moms have attended, including 24% expecting mothers. Additional attendees include 13 family support persons, 23 guest speakers, and 12 from community outreach programs. Qualitative feedback from participants has been uniformly positive. Breastfeeding initiation rates have increased 12% in the initial neighborhood. Conclusions: Harnessing strength within the local community, Champion Breastfeeding Moms have successfully launched AMEN breastfeeding support groups in under-resourced African American urban neighborhoods, helping more mothers reach their breastfeeding goals.

Mobocertinib (TAK-788): A Targeted Inhibitor of EGFR Exon 20 Insertion Mutants in Non-Small Cell Lung Cancer.

Most EGFR exon 20 insertion (EGFRex20ins) driver mutations in non-small cell lung cancer (NSCLC) are insensitive to approved EGFR tyrosine kinase inhibitors (TKI). To address the limitations of existing therapies targeting EGFR-mutated NSCLC, mobocertinib (TAK-788), a novel irreversible EGFR TKI, was specifically designed to potently inhibit oncogenic variants containing activating EGFRex20ins mutations with selectivity over wild-type EGFR. The in vitro and in vivo activity of mobocertinib was evaluated in engineered and patient-derived models harboring diverse EGFRex20ins mutations. Mobocertinib inhibited viability of various EGFRex20ins-driven cell lines more potently than approved EGFR TKIs and demonstrated in vivo antitumor efficacy in patient-derived xenografts and murine orthotopic models. These findings support the ongoing clinical development of mobocertinib for the treatment of EGFRex20ins-mutated NSCLC. SIGNIFICANCE: No oral EGFR-targeted therapies are approved for EGFR exon 20 insertion (EGFRex20ins) mutation-driven NSCLC. Mobocertinib is a novel small-molecule EGFR inhibitor specifically designed to target EGFRex20ins mutants. Preclinical data reported here support the clinical development of mobocertinib in patients with NSCLC with EGFR exon 20 insertion mutations.See related commentary by Pacheco, p. 1617.This article is highlighted in the In This Issue feature, p. 1601.

Nasal DNA methylation is associated with childhood asthma.

AIM: We aim to study DNA methylation (DNAm) variations associated with childhood asthma. METHODS: Nasal DNAm was compared between sibling pairs discordant for asthma, 29 sib pairs for genome-wide association studies and 54 sib pairs for verification by pyrosequencing. Associations of methylation with asthma symptoms, allergy and environmental exposures were evaluated. In vitro experiments and functional genomic analyses were performed to explore biologic relevance. RESULTS: Three CpGs were associated with asthma. cg14830002 was associated with allergies in nonasthmatics. cg23602092 was associated with asthma symptoms. cg14830002 and cg23602092 were associated with traffic-related air pollution exposure. Nearby genes were transcriptionally regulated by diesel exhaust, house dust mite and 5-aza-2'-deoxycytidine. Active chromatin marks and transcription factor binding were found around these sites. CONCLUSION: We identified novel DNAm variations associated with childhood asthma and suggested new disease-contributing epigenetic mechanisms.

IL-17A enhances IL-13 activity by enhancing IL-13-induced signal transducer and activator of transcription 6 activation.

BACKGROUND: Increased IL-17A production has been associated with more severe asthma; however, the mechanisms whereby IL-17A can contribute to IL-13-driven pathology in asthmatic patients remain unclear. OBJECTIVE: We sought to gain mechanistic insight into how IL-17A can influence IL-13-driven responses. METHODS: The effect of IL-17A on IL-13-induced airway hyperresponsiveness, gene expression, mucus hypersecretion, and airway inflammation was assessed by using in vivo models of IL-13-induced lung pathology and in vitro culture of murine fibroblast cell lines and primary fibroblasts and human epithelial cell lines or primary human epithelial cells exposed to IL-13, IL-17A, or both. RESULTS: Compared with mice given intratracheal IL-13 alone, those exposed to IL-13 and IL-17A had augmented airway hyperresponsiveness, mucus production, airway inflammation, and IL-13-induced gene expression. In vitro, IL-17A enhanced IL-13-induced gene expression in asthma-relevant murine and human cells. In contrast to the exacerbating influence of IL-17A on IL-13-induced responses, coexposure to IL-13 inhibited IL-17A-driven antimicrobial gene expression in vivo and in vitro. Mechanistically, in both primary human and murine cells, the IL-17A-driven increase in IL-13-induced gene expression was associated with enhanced IL-13-driven signal transducer and activator of transcription 6 activation. CONCLUSIONS: Our data suggest that IL-17A contributes to asthma pathophysiology by increasing the capacity of IL-13 to activate intracellular signaling pathways, such as signal transducer and activator of transcription 6. These data represent the first mechanistic explanation of how IL-17A can directly contribute to the pathogenesis of IL-13-driven pathology.

Acquisition of a single EZH2 D1 domain mutation confers acquired resistance to EZH2-targeted inhibitors.

Although targeted therapies have revolutionized cancer treatment, overcoming acquired resistance remains a major clinical challenge. EZH2 inhibitors (EZH2i), EPZ-6438 and GSK126, are currently in the early stages of clinical evaluation and the first encouraging signs of efficacy have recently emerged in the clinic. To anticipate mechanisms of resistance to EZH2i, we used a forward genetic platform combining a mutagenesis screen with next generation sequencing technology and identified a hotspot of secondary mutations in the EZH2 D1 domain (Y111 and I109). Y111D mutation within the WT or A677G EZH2 allele conferred robust resistance to both EPZ-6438 and GSK126, but it only drove a partial resistance within the Y641F allele. EZH2 mutants required histone methyltransferase (HMT) catalytic activity and the polycomb repressive complex 2 (PRC2) components, SUZ12 and EED, to drive drug resistance. Furthermore, D1 domain mutations not only blocked the ability of EZH2i to bind to WT and A677G mutant, but also abrogated drug binding to the Y641F mutant. These data provide the first cellular validation of the mechanistic model underpinning the oncogenic function of WT and mutant EZH2. Importantly, our findings suggest that acquired-resistance to EZH2i may arise in WT and mutant EZH2 patients through a single mutation that remains targetable by second generation EZH2i.

Stapled α-helical peptide drug development: a potent dual inhibitor of MDM2 and MDMX for p53-dependent cancer therapy.

Stapled α-helical peptides have emerged as a promising new modality for a wide range of therapeutic targets. Here, we report a potent and selective dual inhibitor of MDM2 and MDMX, ATSP-7041, which effectively activates the p53 pathway in tumors in vitro and in vivo. Specifically, ATSP-7041 binds both MDM2 and MDMX with nanomolar affinities, shows submicromolar cellular activities in cancer cell lines in the presence of serum, and demonstrates highly specific, on-target mechanism of action. A high resolution (1.7-Å) X-ray crystal structure reveals its molecular interactions with the target protein MDMX, including multiple contacts with key amino acids as well as a role for the hydrocarbon staple itself in target engagement. Most importantly, ATSP-7041 demonstrates robust p53-dependent tumor growth suppression in MDM2/MDMX-overexpressing xenograft cancer models, with a high correlation to on-target pharmacodynamic activity, and possesses favorable pharmacokinetic and tissue distribution properties. Overall, ATSP-7041 demonstrates in vitro and in vivo proof-of-concept that stapled peptides can be developed as therapeutically relevant inhibitors of protein-protein interaction and may offer a viable modality for cancer therapy.

Detection of articular pathology of the distal aspect of the third metacarpal bone in thoroughbred racehorses: comparison of radiography, computed tomography and magnetic resonance imaging.

OBJECTIVE: To compare digital radiography (DR), computed tomography (CT), and magnetic resonance imaging (MRI) for detection of pathology of the distal aspect of the third metacarpal bone (MC3) and to assess whether arthrography would improve detection of articular cartilage or subchondral bone cracking. STUDY DESIGN: Cross-sectional study. SAMPLE POPULATION: Limb specimens from 17 Thoroughbred horses after catastrophic injury and 4 age-matched control horses. METHODS: Standard DR, CT, and MRI images of the metacarpophalangeal joint were acquired before and after iohexol injection. Pathologic features detected with imaging and on visual inspection of cartilage and subchondral bone of the distal aspect of MC3 were graded. Imaging observations were compared with pathologic abnormalities. RESULTS: Inspection revealed obvious changes in the cartilage and subchondral bone surfaces in Thoroughbreds. Both CT and MRI were superior to DR for detection of subchondral bone pathology. Cracking of cartilage was not detected by any imaging modality. Signal changes associated with cartilage loss and development of repair tissue were evident on MRI in 9/19 cases. There was significant correlation (P < .05) between subchondral bone pathology detected on both CT and MRI, and cartilage pathology on gross examination. Contrast arthrography did not improve the detection of articular cartilage or subchondral plate cracking. CONCLUSION: Both CT and MRI are superior to DR for detection of subchondral bone pathology, but underestimate the extent of joint adaptation and pathologic damage. MRI was able to detect cartilage degeneration.

A soluble activin receptor type IIb prevents the effects of androgen deprivation on body composition and bone health.

Androgen deprivation, a consequence of hypogonadism, certain cancer treatments, or normal aging in men, leads to loss of muscle mass, increased adiposity, and osteoporosis. In the present study, using a soluble chimeric form of activin receptor type IIB (ActRIIB) we sought to offset the adverse effects of androgen deprivation on muscle, adipose tissue, and bone. Castrated (ORX) or sham-operated (SHAM) mice received either TBS [vehicle-treated (VEH)] or systemic administration of ActRIIB-mFc, a soluble fusion protein comprised of a form of the extracellular domain of ActRIIB fused to a murine IgG2aFc subunit. In vivo body composition imaging demonstrated that ActRIIB-mFc treatment results in increased lean tissue mass of 23% in SHAM mice [19.02 +/- 0.42 g (VEH) versus 23.43 +/- 0.35 g (ActRIIB-mFc), P < 0.00001] and 26% in ORX mice [15.59 +/- 0.26 g (VEH) versus 19.78 +/- 0.26 g (ActRIIB-mFc), P < 0.00001]. Treatment also caused a decrease in adiposity of 30% in SHAM mice [5.03 +/- 0.48 g (VEH) versus 3.53 +/- 0.19 g (ActRIIB-mFc), NS] and 36% in ORX mice [7.12 +/- 0.53 g (VEH) versus 4.57 +/- 0.28 g (ActRIIB-mFc), P < 0.001]. These changes were also accompanied by altered serum levels of leptin, adiponectin, and insulin, as well as by prevention of steatosis (fatty liver) in ActRIIB-mFc-treated ORX mice. Finally, ActRIIB-mFc prevented loss of bone mass in ORX mice as assessed by whole body dual x-ray absorptiometry and micro-computed tomography of proximal tibias. The data demonstrate that treatment with ActRIIB-mFc restored muscle mass, adiposity, and bone quality to normal levels in a mouse model of androgen deprivation, thereby alleviating multiple adverse consequences of such therapy.

Efficacy of leflunomide for treatment of immune-mediated polyarthritis in dogs: 14 cases (2006-2008).

OBJECTIVE: To evaluate efficacy and adverse effects of leflunomide for the treatment of naturally occurring immune-mediated polyarthritis (IMPA) in dogs. DESIGN: Retrospective case series. ANIMALS: 14 dogs with cytologically confirmed IMPA. PROCEDURES: Medical records were used to identify dogs with a diagnosis of IMPA that were treated with leflunomide. Signalment, radiographic findings, laboratory data, dosage of leflunomide, treatment duration, treatment response, and occurrence of adverse effects were determined from medical records. RESULTS: Mean +/- SD initial dosage of leflunomide was 3.0 +/- 0.5 mg/kg (1.4 +/- 0.2 mg/lb) PO once daily. Treatment duration for the initial starting dosage ranged from 1 to 6 weeks. Of the 14 dogs treated with leflunomide, 8 had complete resolution of clinical signs of IMPA initially, 5 had partial response to treatment, and 1 had minimal response to treatment. Adverse effects from treatment with leflunomide were not observed during the treatment period. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of leflunomide was a safe and effective alternative to oral administration of corticosteroids for treatment of IMPA in dogs. On the basis of findings in this study, a starting dosage for leflunomide of 3 to 4 mg/kg (1.4 to 1.8 mg/lb) PO once daily for at least 6 weeks before making dose adjustments is recommended. Dose adjustments should be based on cytologic evaluation of synovial fluid and clinical signs of IMPA. Hematologic variables, serum biochemical analysis results, and clinical signs of IMPA should be monitored for evidence of adverse effects to treatment with leflunomide.

Seasonal variation in detection of bacterial DNA in arthritic stifle joints of dogs with cranial cruciate ligament rupture using PCR amplification of the 16S rRNA gene.

An underappreciated cause and effect relationship between environmental bacteria and arthritis may exist. Previously, we found that stifle arthritis in dogs was associated with the presence of environmental bacteria within synovium. Cranial cruciate ligament rupture (CCLR) is often associated with stifle arthritis in dogs. We now wished to determine whether seasonal variation in detection of bacterial material may exist in affected dogs, and to also conduct analyses of both synovium and synovial fluid. We also wished to analyze a larger clone library of the 16S rRNA gene to further understanding of the microbial population in the canine stifle. Synovial biopsies were obtained from 117 affected dogs from January to December 2006. Using PCR, synovium and synovial fluid were tested for Borrelia burgdorferi and Stenotrophomonas maltophilia DNA. Broad-ranging 16S rRNA primers were also used and PCR products were cloned and sequenced for bacterial identification. Overall, 41% of arthritic canine stifle joints contained bacterial DNA. Detection of bacterial DNA in synovial fluid samples was increased, when compared with synovium (p<0.01). Detection rates were highest in the winter and spring and lowest in the summer period, suggesting environmental factors influence the risk of translocation to the stifle. Organisms detected were predominately Gram's negative Proteobacteria, particularly the orders Rhizobiales (32.8% of clones) and Burkholderiales (20.0% of clones), usually as part of a polymicrobial population. PCR-positivity was inversely correlated with severity of arthritis assessed radiographically and with dog age. Bacterial translocation to the canine stifle may be associated with changes to the indoor environment.

Mechanical loading of a long bone induces plasticity in sensory input to the central nervous system.

Although the skeleton is extensively innervated by sensory nerves, the importance of this innervation to skeletal physiology is unclear. Neuronal connectivity between limbs is little studied and likely underestimated. In this study, we examined the effect of bone loading on spinal plasticity in young male Sprague-Dawley rats, using end-loading of the ulna and transynaptic tracing with the Bartha pseudorabies virus (PRV). PRV was inoculated onto the periosteum of the right ulna after 10 days of adaptation to a single period of cyclic loading of the right ulna (1,500 cycles of load at 4 Hz, initial peak strain of -3,750 micro epsilon). We found that neuronal circuits connect the sensory innervation of right thoracic limb to all other limbs, as PRV was detectable in the dorsal root ganglia (DRG) of left and right brachial and lumbosacral intumescences. We also found that mechanical loading of the right ulna induced plasticity in the spinal cord, with significant augmentation of the connectivity between limbs, as measured by PRV translocation. Within the spinal cord, PRV was predominantly found adjacent to the central canal and in the dorsal horns, suggesting that plasticity in cross-talk between limbs is likely a consequence of dendritic growth, and enhanced connectivity of propriospinal interneurons. In conclusion, the data clearly demonstrate that the innervation of the skeleton exhibits plasticity in response to loading events, suggesting the existence of a dynamic control system that may be of regulatory importance during functional skeletal adaptation.