Multiple Family Members Research Articles

Strategies for managing group caregiving following hip-fracture surgery among family members: A grounded theory study.

Family members in many countries often share caregiving responsibilities for an older relative recovering from an injury. However, few studies have examined strategies employed when multiple family members provide care for an older relative recovering from hip-fracture surgery. This study aimed to understand family group caregiving strategies when two or more family members provide caregiving for an older relative recovering from hip-fracture surgery. This study used a grounded theory design. Semistructured interviews were conducted over 1 year with 13 Taiwanese family caregivers from five families. Caregivers shared caregiving responsibilities for an older relative (62-92 years of age) recovering from hip-fracture surgery. Transcribed interviews were analysed using open, axial and selective coding. The core category describing caregiving among family members was 'Preventive Group Management: strategies for family group caregiving'. Three strategies were employed: explicit division of labour (two stem/patriarchal families and one older two-generation/democratic family); disconnected caregiving (one nuclear/noncommunicative family) and patriarchal caregiving (one extended/traditional Chinese family). Strategies reflected family type, structure, cultural values, communication patterns and available outside support. Components of family group caregiving involved family type's division of labour, approaches to caregiving and implementation challenges and allowed family caregivers to maximise safety and stability and prevent harmful events during their relative's recovery from surgery. There was no one-size-fits-all approach for the strategies of family group caregiving. Components of Preventive Group Management varied with family type, cultural values, communication patterns and available outside support. Healthcare professionals should be sensitive to the dynamics of family caregivers. Enhance group management for family caregivers by developing interventions to optimize collaboration, thereby better addressing the needs of older adults recovering from hip fracture surgery.

Laboratory Selection in Germline Genetic Testing: Laboratory Science Matters.

There are multiple laboratories that offer germline genetic testing, and it can be difficult to discern which one to use for testing. Some laboratories have more comprehensive analysis techniques and capability, which increases the accuracy of testing. The ordering provider has a responsibility to select the appropriate laboratory with technologic capability for the needed testing, inform the laboratory of prior testing results in the patient and family so known familial variants have targeted testing, and use appropriate terminology and nomenclature when communicating information to other healthcare professionals, patients, and families. This report presents a case illustrating the potential errors that can occur when a provider selects a laboratory that lacks the capacity to detect certain pathogenic variants, such as large deletions and duplications. False-negative germline testing results lead to missed opportunities in prevention and early detection for not only the patient but often multiple family members, which may lead to psychosocial distress and late-detected cancers. This case highlights the complexities of genetic care and why management by a genetics professional can facilitate more fiscally responsible care, appropriate genetic testing, and comprehensive care for all family members at risk.

Staff and Proxy Views of Multiple Family Member Involvement in Decision Making for Nursing Home Residents With Advanced Dementia.

Decision making for nursing home (NH) residents with Alzheimer disease and related dementias often involves input from multiple family members and NH staff to address goals of care at the end of life. Using data from the Assessment of Disparities and Variation for Alzheimer's disease Nursing home Care at End of life research study, a secondary analysis of qualitative data was conducted involving interviews of 144 NH staff and 44 proxies in 14 NHs to examine the perspectives of NH staff and proxies for NH residents with Alzheimer disease and related dementias on the involvement of multiple family members in decision making about end-of-life care decisions. Interviews took place between 2018 and 2021. Nursing home staff and proxies had differing perspectives of the involvement of multiple family members in decision making, with NH staff primarily viewing families as a source of conflict, whereas proxies viewed families as a source of support. Nursing home staff also had differing opinions of their role with families; some attempted to ameliorate conflict, and some did not get involved. Some NH staff felt that Black families had more conflict than White families, indicating unacceptable bias and stereotyping of Black families by NH staff. These findings suggest training and education is needed for NH staff to facilitate better communication with families and to support proxies in end-of-life decision making to address goals of care for NH residents with Alzheimer disease and related dementias.

Designing a Dyad-Based Digital Health Intervention to Reduce Sedentary Time in Black Breast Cancer Survivors and Their First-degree Relatives: Human-Centered Design Study.

Breast cancer, the most commonly diagnosed cancer and second leading cause of cancer-related death in women in the United States, disproportionately affects women from minoritized or low socioeconomic backgrounds. The average woman has an approximately 12% lifetime risk of developing breast cancer. Lifetime risk nearly doubles if a woman has a first-degree relative with breast cancer, and the risk increases as multiple family members are affected. Decreasing sedentary behaviors through moving more and sitting less reduces breast cancer risk and improves outcomes for cancer survivors and healthy adults. Digital health solutions, such as mobile apps that are culturally appropriate, designed with input from the target audience, and include social support, are effective at improving health behaviors. This study aimed to develop and evaluate the usability and acceptability of a prototype app designed with a human-centered approach to promote moving more and sitting less in Black breast cancer survivors and their first-degree relatives (parent, child, or sibling). This 3-phase study consisted of app development, user testing, and evaluation of user engagement and usability. Key community stakeholders were engaged in the first 2 (qualitative) phases to provide input into developing the prototype app (MoveTogether). After development and user testing, a usability pilot was conducted. Participants were adult breast cancer survivors who identified as Black and agreed to participate with a relative. Participants used the app and a step-tracking watch for 4 weeks. App components included goal setting and reporting, reminders, dyad messaging, and educational resources. Usability and acceptability were assessed with a questionnaire that included the System Usability Scale (SUS) and semistructured interviews. Data were analyzed with descriptive statistics and content analysis. Participants in the usability pilot (n=10) were aged 30 to 50 years (6/10, 60%), not married (8/10, 80%), and college graduates (5/10, 50%). The app was used on average 20.2 (SD 8.9) out of 28 days-SUS score of 72 (range 55-95)-and 70% (7/10) agreed that the app was acceptable, helpful, and gave them new ideas. Additionally, 90% (9/10) found the dyad component helpful and would recommend the app to friends. Qualitative findings suggest that the goal-setting feature was helpful and that the dyad partner (buddy) provided accountability. Participants were neutral regarding the cultural appropriateness of the app. The MoveTogether app and related components were acceptable for promoting moving more in dyads of breast cancer survivors and their first-degree relatives. The human-centered approach, which involved engaging community members in the development, is a model for future technology development work. Future work should be done to further develop the intervention based on the findings and then test its efficacy to improve sedentary behavior while considering culturally informed strategies for adoption and implementation within the community. ClinicalTrials.gov NCT05011279; https://clinicaltrials.gov/ct2/show/NCT05011279.

Adolescent difficulties during parental deployment and anxiety: A focus on measurement and family processes

AbstractApproximately 60% of deployed service members leave behind immediate family members, and although military families tend to be adaptive and resilient, evidence suggests that deployments are challenging and difficulties can arise during transitions and family separation, especially for adolescents. Grounded in the family attachment network model and the ABC‐X model of family stress, the current study utilized a sample of 204 military families with an active‐duty father, civilian mother and adolescent and examined parents' perceptions of adolescents' difficulties during deployment in relation to all three family members' perceptions of the adolescents' mental health (i.e., anxiety symptoms) following deployment. First, analyses of measurement invariance indicated that service members and civilian parents were generally reporting on the same underlying construct of their adolescents' difficulties during parental deployment. Next, a structural equation model demonstrated considerable overlap in service member and civilian parent reports of their adolescents' difficulties during a parental deployment (r = 0.47). Finally, both parents' perceptions of adolescent difficulties during parental deployment were related to their own perceptions of the adolescent's current anxiety but not to the adolescents' reports of their own anxiety symptoms or to the other parent's report of the adolescents' anxiety symptoms. Findings provide support for utilizing these theories in combination, such that disruptions to the family system, and the attachment relationships within that system, in one stage of the deployment cycle, may imply that there are implications for individual‐level functioning, namely, anxiety, in the next stage of the deployment cycle. Findings also underscore the importance of examining our measurement tools and collecting data from multiple family members to understand family processes.

The deubiquitylating enzyme USP35 restricts regulated cell death to promote survival of renal clear cell carcinoma.

The ubiquitin-proteasome system governs a wide spectrum of cellular events and offers therapeutic opportunities for pharmacological intervention in cancer treatment. Renal clear cell carcinoma represents the predominant histological subtype and accounts for the majority of cancer death related to kidney malignancies. Through a systematic survey in the association of human ubiquitin-specific proteases with patient prognosis of renal clear cell carcinoma and subsequent phenotypic validation, we uncovered the tumor-promoting role of USP35. Biochemical characterizations confirmed the stabilizing effects of USP35 towards multiple members of the IAP family in an enzymatic activity-dependent manner. USP35 silencing led to reduced expression levels of IAP proteins, which were accompanied with increased cellular apoptosis. Further transcriptomic analysis revealed that USP35 knockdown affected the expression levels of NRF2 downstream transcripts, which were conferred by compromised NRF2 abundance. USP35 functions to maintain NRF2 levels by catalyzing its deubiquitylation and thus antagonizing degradation. NRF2 reduction imposed by USP35 silencing rendered renal clear cell carcinoma cells increased sensitivity to ferroptosis induction. Finally, induced USP35 knockdown markedly attenuated xenograft formation of renal clear cell carcinoma in nude mice. Hence, our findings reveal a number of USP35 substrates and uncover the protecting roles of USP35 against both apoptosis and ferroptosis in renal clear cell carcinoma.

A rising tide lifts all MBOATs: recent progress in structural and functional understanding of membrane bound O-acyltransferases.

Acylation modifications play a central role in biological and physiological processes. Across a range of biomolecules from phospholipids to triglycerides to proteins, introduction of a hydrophobic acyl chain can dramatically alter the biological function and cellular localization of these substrates. Amongst the enzymes catalyzing these modifications, the membrane bound O-acyltransferase (MBOAT) family occupies an intriguing position as the combined substrate selectivities of the various family members span all three classes of these biomolecules. MBOAT-dependent substrates are linked to a wide range of health conditions including metabolic disease, cancer, and neurodegenerative disease. Like many integral membrane proteins, these enzymes have presented challenges to investigation due to their intractability to solubilization and purification. However, over the last several years new solubilization approaches coupled with computational modeling, crystallography, and cryoelectron microscopy have brought an explosion of structural information for multiple MBOAT family members. These studies enable comparison of MBOAT structure and function across members catalyzing modifications of all three substrate classes, revealing both conserved features amongst all MBOATs and distinct architectural features that correlate with different acylation substrates ranging from lipids to proteins. We discuss the methods that led to this renaissance of MBOAT structural investigations, our new understanding of MBOAT structure and implications for catalytic function, and the potential impact of these studies for development of new therapeutics targeting MBOAT-dependent physiological processes.

Clinical Pearls

During the pandemic, extracorporeal membrane oxygenation (ECMO) was instituted for patients with COVID-19 hypoxemia refractory to mechanical ventilation support. Although some patients had favorable outcomes, many who survived hospitalization were left with significant functional disability. Palliative care consultation with family members of ECMO patients has been studied, but the focus has primarily been on end-of-life care and not on overall care expectations.Siddiqui and colleagues implemented an early, scripted palliative care intervention for families of patients with COVID-19 receiving ECMO. After interviewing families, most within 24 to 72 hours of ECMO initiation, they identified these themes: Social isolation and related grief of not being at the patient’s bedsideHelping patients’ children adjust to a new normalCoping with multiple family members suffering from COVID-19Importance of faith and spiritualityNeed for hope and gratitudeFutility of prolonged ECMO therapyPalliative care has expanded to care for patients at all stages of chronic illness. The study findings support early family engagement and assistance for those dealing with COVID-19 and similar illnesses.See Article, pp 166-174Workplace stress can contribute to negative consequences for both staff and patients. Caring for critically ill patients adds additional stressors for intensive care nurses, which were further exacerbated by the COVID-19 pandemic. Although some interventions exist for nurse support, none focus on nurses taking time out to meet their own needs.Smith and colleagues assessed nurses’ perceptions of their stress and the usability of a Lavender Lounge—a quiet room with items and activities available to help them destress. They found The very factors that lead to burnout were the ones perceived as barriers to use of the Lavender Lounge. The authors suggest an approach that includes organizational change that supports reduced staff workload.See Article, pp 198-204Thirst is a complex symptom that affects between 49% and 98% of adult patients. Unrelieved thirst can become a distressing experience that overshadows other ailments. Although nurses can alleviate the distress caused by thirst, there are no standard protocols for accurate assessment and evaluation.Ciftci and colleagues developed and tested the Thirst Discomfort Scale to standardize measurement of thirst in adult patients. They found Although further study with other types of patients is warranted, the Thirst Discomfort Scale can be used to assess and evaluate the symptom of thirst in adult patients.See Article, pp 176-183Pressure injury (PI) prevention and treatment guidelines have historically been geared toward adult patients and adapted by pediatric providers. However, children with congenital heart disease (CHD) have unique factors that increase their susceptibility to PIs.To prevent PIs, Shields and colleagues explored the specific risk factors of the care and management of PIs in pediatric patients with CHD. They found This study adds to the limited literature on key risk factors for pediatric patients with CHD developing PIs. Clinicians providing care for these patients should use this knowledge to decrease the incidence of PIs.See Article, pp 216-220

Discovery of a molecular clock that controls CD8+ T cell function and exhaustion

Abstract During cancer and chronic viral infections, the persistence of antigen progressively causes CD8+ T cells to differentiate into a dysfunctional PD1+ “exhausted” state, with reduced production of inflammatory cytokines relative to effector cells that form during acute infections. Antigen and costimulation signals activate kinase cascades to induce distinct T cell transcription programs, but how T cells distinguish acute and chronic signals to program the exhausted or effector transcriptional states remains poorly understood. We found that members of the protein kinase C (PKC) family function together as a “molecular clock,” sensing acute or chronic agonism to drive distinct transcriptional programs. Continuous stimulation of PKC induces many features of T cell exhaustion, including a loss of production of the cytokines IFNγ and TNF, upregulation of inhibitory receptors and TOX, and altered expression of the proteins in the AP-1 family. Mechanistically, CD8+ T cells express several different PKC proteins, and chronic agonism of PKC leads to degradation of multiple family members and selective maintenance of only one PKC protein, PKC-η. This “PKC switch” alters downstream signaling to support the transcriptional reprogramming of T cells into a terminally exhausted state. In summary, continuous signaling through PKCs causes changes in the output from these kinases initially at the protein level, driving transcriptional changes downstream of PKC targets in the AP-1 transcription factor family and thus allowing further widespread transcriptional and functional changes that characterize T cell exhaustion. Supported by a Damon Runyon Cancer Research Fellowship (DRG-2358-19) and an NIH training grant, T32-CA009370.

TCRvβ-CART therapy mediates high-precision targeting of malignant T-cell clones.

Peripheral T-cell lymphomas (PTCLs) are a heterogeneous group of lymphoid malignancies associated with poor prognosis due to ineffective treatment options and high rates of relapse. The success of chimeric antigen receptor T-cell (CART) therapy for certain hematologic malignancies makes it an attractive treatment option for PTCLs. However, shared expression of potential target antigens by both malignant and healthy T cells poses a challenge. Current prospective CART approaches cause a high degree of on-target, off-tumor activity, resulting in fratricide during CART expansion, depletion of healthy Tcells invivo, and immune compromise in the patient. To limit off-tumor targeting, we sought to develop a CART platform specific for a given T-cell receptor vβ (TCRvβ) family thatwould endow CAR-modified T cells with the ability to mediate lysis of the clonal malignant population while preserving the majority of healthy T cells. Here, CAR constructs specific for multiple TCRvβ family members were designed and validated. Our results demonstrate that TCRvβ-family-specific CARTs (TCRvβ-CARTs) recognize and kill TCRvβ-expressing target cells. This includes specific self-depletion of the targeted cell subpopulation in the CART product and lysis of cell lines engineered to express a target TCRvβ family. Furthermore, TCRvβ-CARTs eliminated the dominant malignant TCRvβ clonein 2 patient samples. Finally, in immunodeficient mice, TCRvβ-CARTs eradicated malignant cells in a TCRvβ-dependent manner. Importantly, the nontargeted TCRvβ families were spared in all cases. Thus, TCRvβ-CART therapy provides a potential option for high-precision treatment of PTCL with limited healthy T-cell depletion.

Four Looks to a Gene (P1-8.013)

<h3>Objective:</h3> We present a family with varied clinical presentations of a genetically confirmed muscular dystrophy. <h3>Background:</h3> A 41-year-old male presented with several years of muscle fatigue, pain and twitching across his chest and proximal muscles. He was diagnosed with neuronal hyperexcitability. Exam showed normal muscle strength, but there were visible rippling in proximal muscles. Cardiac workup was unrevealing. EMG showed nonspecific myopathic features and CPK was in the 400’s. His 9-year-old son has been diagnosed with muscular dystrophy and two daughters has elevated CPKs. Our patient was genetically confirmed to have a heterozygous C.80G&gt;A(p.Arg27Gln) mutation in the Caveolin-3 gene via Invitae muscular dystrophy testing. This missense mutation causes an amino acid change from Arg to Gln in the Caveolin 3 protein, which leads to a decrease of Cav-3 protein expression in skeletal muscle tissue. It has an autosomal dominant inheritance pattern. <h3>Design/Methods:</h3> Not applicable <h3>Results:</h3> Mutations in the Caveolin-3 gene have been described in a broad spectrum of clinical presentations including limb girdle muscular dystrophy, rippling muscle disease, idiopathic persistent hyperCKemia, and distal myopathy. Muscular dystrophy can present with a wide variety of phenotypes and there is even variation within the same gene mutations, as is the case in caveolin 3 mutations. This may lead to delay in diagnosis and supportive treatments that a family may require. The Invitae muscular dystrophy panel can be helpful in expediting diagnosis for multiple family members. It is also important to remember that muscular dystrophy can mimic muscle hyperexcitability syndrome, as was the case in our patient with rippling muscle disease. <h3>Conclusions:</h3> Muscular dystrophy can present with distinct phenotypic variations. A good family history and awareness of clinical symptoms can be helpful for diagnosis. CAV3 mutations cause LGMD1C which can present with at least four different phenotypes. <b>Disclosure:</b> Dr. Rahimi has nothing to disclose. Dr. Memar Ardestani has nothing to disclose. Dr. Prabhu has nothing to disclose. Dr. Undavia has nothing to disclose.

Synthesis of active cytokinins mediated by LONELY GUY is associated with cell production during early fruit growth in peach [Prunus persica (L.) Batsch

Early fruit growth in peach is characterized by cell production. Cytokinins have established roles in regulating cell division and may regulate cell production during early fruit growth. However, the role of active cytokinins and regulation of their metabolism are not well characterized in the peach fruit. In this study, fruit growth parameters, concentrations of active cytokinin bases and a cytokinin riboside, and expression of three key cytokinin metabolism-related gene families were determined during early fruit growth. Early fruit growth was associated with intensive cell production until around 40 days after full bloom. During the early stages of this period, trans-zeatin (tZ), isopentenyladenine (iP), dihydrozeatin (DHZ) and tZ-riboside (tZR), displayed higher abundance which declined rapidly by 3.5- to 16-fold during the later stages. Changes in concentration of active cytokinin bases were consistent with roles for them in regulating cell production. Expression analyses of members of cytokinin biosynthesis-related gene families, ISOPENTENYL TRANSFERASE (IPT) and LONELY GUY (LOG), further indicated that mechanisms of synthesis of cytokinin metabolites and their activation are functional within the fruit pericarp. Changes in expression of multiple members of the LOG family paralleled changes in active cytokinin concentrations. Specifically, transcript abundance of LOG3 and LOG8 were correlated with concentrations of tZ, and iP and DHZ, respectively, suggesting that the direct activation pathway is an important route for active cytokinin base synthesis during early fruit development. Transcript abundance of two CYTOKININ OXIDASE (CKX) genes, CKX1 and CKX2, was consistent with roles in cytokinin catabolism during later stages of early fruit growth. Together, these data support a role for active cytokinins synthesized in the fruit pericarp in regulating early fruit growth in peach.

ECT9 condensates with ECT1 and regulates plant immunity.

Mounting an efficient defense against pathogens requires RNA binding proteins (RBPs) to regulate immune mRNAs transcription, splicing, export, translation, storage, and degradation. RBPs often have multiple family members, raising the question of how they coordinate to carry out diverse cellular functions. In this study, we demonstrate that EVOLUTIONARILY CONSERVED C-TERMINAL REGION 9 (ECT9), a member of the YTH protein family in Arabidopsis, can condensate with its homolog ECT1 to control immune responses. Among the 13 YTH family members screened, only ECT9 can form condensates that decrease after salicylic acid (SA) treatment. While ECT1 alone cannot form condensates, it can be recruited to ECT9 condensates in vivo and in vitro. Notably, the ect1/9 double mutant, but not the single mutant, exhibits heightened immune responses to the avirulent pathogen. Our findings suggest that co-condensation is a mechanism by which RBP family members confer redundant functions.

Modulating auxin response stabilizes tomato fruit set.

Fruit formation depends on successful fertilization and is highly sensitive to weather fluctuations that affect pollination. Auxin promotes fruit initiation and growth following fertilization. Class A auxin response factors (Class A ARFs) repress transcription in the absence of auxin and activate transcription in its presence. Here, we explore how multiple members of the ARF family regulate fruit set and fruit growth in tomato (Solanum lycopersicum) and Arabidopsis thaliana, and test whether reduction of SlARF activity improves yield stability in fluctuating temperatures. We found that several tomato Slarf mutant combinations produced seedless parthenocarpic fruits, most notably mutants deficient in SlARF8A and SlARF8B genes. Arabidopsis Atarf8 mutants deficient in the orthologous gene had less complete parthenocarpy than did tomato Slarf8a Slarf8b mutants. Conversely, Atarf6 Atarf8 double mutants had reduced fruit growth after fertilization. AtARF6 and AtARF8 likely switch from repression to activation of fruit growth in response to a fertilization-induced auxin increase in gynoecia. Tomato plants with reduced SlARF8A and SlARF8B gene dosage had substantially higher yield than the wild type under controlled or ambient hot and cold growth conditions. In field trials, partial reduction in the SlARF8 dose increased yield under extreme temperature with minimal pleiotropic effects. The stable yield of the mutant plants resulted from a combination of early onset of fruit set, more fruit-bearing branches and more flowers setting fruits. Thus, ARF8 proteins mediate the control of fruit set, and relieving this control with Slarf8 mutations may be utilized in breeding to increase yield stability in tomato and other crops.

Abstract 4042: Role of individual HER family members and pan-HER targeting treatment strategy in NRG1 fusion positive cancer

Abstract NRG1 gene fusions are rare, clinically actionable somatic alterations identified in 0.1% of all tumors. Previous studies have demonstrated that NRG1 fusions signal through ERBB/HER family members and that HER2 inhibition has anti-tumor activity in NRG1 fusion-driven cancers. However, NRG1 can also bind to HER4 in addition to HER3, and the contribution of individual HER family members in tumor cells with NRG1 fusions has not been fully elucidated. To determine the role of individual HER family members in NRG1 fusion positive cancer cells, we engineered Ba/F3 cells to express various HER family members along with NRG1 fusions. Expression of NRG1 fusions in combination with EGFR/HER3, HER2/HER4 or HER4 induced cellular transformation as measured by IL-3 independent growth. These results indicated that in addition to HER2/HER3 signaling, EGFR/HER3, HER4, and to a greater extent, HER2/HER4 signaling, may activate key oncogenic signaling pathways in cells expressing NRG1 fusions. We next assessed whether targeted inhibition of HER family members could inhibit the growth of NRG1 fusion positive cancer cells using our panel of engineered Ba/F3 cells, human tumor cell lines, and xenograft models. Our data indicated that pan-HER tyrosine kinase inhibitors (TKIs), such as poziotinib, were more effective at blocking HER2/HER3, EGFR/HER3 and HER4 signaling in NRG1-fusion expressing cells as compared to TKIs with greater relative specificity for EGFR (erlotinib, lapatinib), HER2 (pyrotinib), EGFR/HER2 (afatinib, dacomitinib, neratinib) or HER2/HER4 (TAS0728,tucatinib). In NRG1 fusion-positive cancer cell lines, poziotinib showed potent anti-tumor cell activity with IC50 values of 0.2 - 0.4 nM. The IC50 values of EGFR/HER2 TKIs were 0.3 - 23 nM, and HER2/HER4 TKIs showed modest activity with IC50 values of 11 - 560 nM. Next, we assessed whether inhibition of HER family dimerization also had activity against tumor cells harboring NRG1 fusions. We observed that pertuzumab, a HER2 antibody which inhibits HER2 dimerization with other HER family members, had improved single agent activity as compared to trastuzumab, a HER2 antibody that inhibits HER2 activation, suggesting crosstalk between HER family members. Moreover, an additive effect was observed when tumor cells were treated with trastuzumab in combination with pertuzumab. Treatment with cetuximab, an antibody that targets EGFR, in combination with trastuzumab and pertuzumab yielded a synergistic effect on tumor cell killing. These data indicate that HER4 and EGFR can play a role in NRG1 fusion-driven signaling through crosstalk with HER2/HER3 and thus, pan-HER/EGFR inhibitors are more effective than EGFR/HER2 or HER2/HER4-selective inhibitors, highlighting the therapeutic potential of targeting multiple members of the HER family in NRG1 fusion driven cancers. Citation Format: Hibiki Udagawa, Monique Nilsson, Junqin He, Alissa Poteete, John Victor Heymach. Role of individual HER family members and pan-HER targeting treatment strategy in NRG1 fusion positive cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4042.

A genome-scale CRISPR tool for targeting multiple gene family members at once.

A genome-scale CRISPR tool for targeting multiple gene family members at once.

SLAMF8, a potential new immune checkpoint molecule, is associated with the prognosis of colorectal cancer

Recently, immune checkpoint inhibitors (ICIs), such as programmed cell death 1 (PD-1) monoclonal antibodies (mAbs), have revolutionized the treatment of malignant tumors. Therefore, the number of studies aiming to screen and identify new immune checkpoint molecules for antitumor immunotherapy is increasing. Signaling lymphocytic activation molecule (SLAM) family members are mainly expressed by and regulate the functions of immune cells. Recent studies have shown that several SLAM family members are involved in the regulation of the tumor immune microenvironment and are promising targets for antitumor immunotherapy. Signaling lymphocytic activation molecule family member 8 (SLAMF8) is a type I cell surface glycoprotein and is encoded on chromosome 1q21. To further illustrate the clinical value of SLAMF8 in colorectal cancer (CRC), we retrospectively analyzed the relationship between SLAMF8 expression and the prognosis of CRC patients and the associations between SLAMF8 expression and the expression levels of other SLAM family members and other classic immune checkpoint molecules using The Cancer Genome Atlas (TCGA) data, RNA sequencing data, tissue immunohistochemistry staining, and systematic follow-up analysis. Here, high SLAMF8 expression was associated with poor overall survival (OS) in CRC. The mRNA expression level of SLAMF8 was positively correlated with the expression levels of multiple classic immune checkpoints and other SLAM family members. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis suggested that the pathways enriched in CRC tissues with high SLAMF8 expression were associated with the regulation of the tumor immune microenvironment.

Large and stable genome edits at the sorghum alpha kafirin locus result in changes in chromatin accessibility and globally increased expression of genes encoding lysine enrichment.

Sorghum is a resilient and widely cultivated grain crop used for feed and food. However, it's grain is deficient in lysine, an essential amino acid. This is due to the primary seed storage proteins, the alpha-kafirins, lacking lysine. It has been observed that reductions in alpha-kafirin protein results in rebalancing of the seed proteome and a corresponding increase in non-kafirin proteins which leads to an increased lysine content. However, the mechanisms underlying proteome rebalancing are unclear. This study characterizes a previously developed gene edited sorghum line, with deletions at the alpha kafirin locus. A single consensus guide RNA leads to tandem deletion of multiple members of the gene family in addition to the small target site mutations in remaining genes. RNA-seq and ATAC-seq were utilized to identify changes in gene expression and chromatin accessibility in developing kernels in the absence of most alpha-kafirin expression. Several differentially accessible chromatin regions and differentially expressed genes were identified. Additionally, several genes upregulated in the edited sorghum line were common with their syntenic orthologues differentially expressed in maize prolamin mutants. ATAC-seq showed enrichment of the binding motif for ZmOPAQUE 11, perhaps indicating the transcription factor's involvement in the kernel response to reduced prolamins. Overall, this study provides a resource of genes and chromosomal regions which may be involved in sorghum's response to reduced seed storage proteins and the process of proteome rebalancing.

Early Guided Palliative Care Communication for Patients With COVID-19 Receiving ECMO.

Extracorporeal membrane oxygenation (ECMO) combined with COVID-19 presents challenges (eg, isolation, anticipatory grief) for patients and families. To (1) describe characteristics and outcomes of patients with COVID-19 receiving ECMO, (2) develop a practice improvement strategy to implement early, semistructured palliative care communication in ECMO acknowledgment meetings with patients' families, and (3) examine family members' experiences as recorded in clinicians' notes during these meetings. Descriptive observation of guided, in-depth meetings with families of patients with COVID-19 receiving ECMO, as gathered from the electronic medical record of a large urban academic medical center. Most meetings were held within 3 days of initiation of ECMO. Forty-three patients received ECMO between March and October 2020. The mean patient age was 44 years; 63% of patients were Hispanic/Latino, 19% were Black, and 7% were White. Documentation of the ECMO acknowledgment meeting was completed for 60% of patients. Fifty-six percent of patients survived to hospital discharge. Family discussions revealed 7 common themes: hope, reliance on faith, multiple family members with COVID-19, helping children adjust to a new normal, visitation restrictions, gratitude for clinicians and care, and end-of-life discussions. Early and ongoing provision of palliative care is feasible and useful for highlighting a range of experiences related to COVID-19. Palliative care is also useful for educating patients and families on the benefits and limitations of ECMO therapy.

Transfer of knowledge from model organisms to evolutionarily distant non-model organisms: The coral Pocillopora damicornis membrane signaling receptome.

With the ease of gene sequencing and the technology available to study and manipulate non-model organisms, the extension of the methodological toolbox required to translate our understanding of model organisms to non-model organisms has become an urgent problem. For example, mining of large coral and their symbiont sequence data is a challenge, but also provides an opportunity for understanding functionality and evolution of these and other non-model organisms. Much more information than for any other eukaryotic species is available for humans, especially related to signal transduction and diseases. However, the coral cnidarian host and human have diverged over 700 million years ago and homologies between proteins in the two species are therefore often in the gray zone, or at least often undetectable with traditional BLAST searches. We introduce a two-stage approach to identifying putative coral homologues of human proteins. First, through remote homology detection using Hidden Markov Models, we identify candidate human homologues in the cnidarian genome. However, for many proteins, the human genome alone contains multiple family members with similar or even more divergence in sequence. In the second stage, therefore, we filter the remote homology results based on the functional and structural plausibility of each coral candidate, shortlisting the coral proteins likely to have conserved some of the functions of the human proteins. We demonstrate our approach with a pipeline for mapping membrane receptors in humans to membrane receptors in corals, with specific focus on the stony coral, P. damicornis. More than 1000 human membrane receptors mapped to 335 coral receptors, including 151 G protein coupled receptors (GPCRs). To validate specific sub-families, we chose opsin proteins, representative GPCRs that confer light sensitivity, and Toll-like receptors, representative non-GPCRs, which function in the immune response, and their ability to communicate with microorganisms. Through detailed structure-function analysis of their ligand-binding pockets and downstream signaling cascades, we selected those candidate remote homologues likely to carry out related functions in the corals. This pipeline may prove generally useful for other non-model organisms, such as to support the growing field of synthetic biology.