Advanced Practice Providers in the Medical Genetics Workforce: A Nationwide Survey.

PURPOSE: This study characterizes the current landscape of genetics advanced practice providers (APPs) in the United States. METHODS: A 35-question survey was emailed to the Genetics APP Listserv in the fall of 2023. Questions represented five domains: demographics, practice, onboarding, compensation, and perceptions. RESULTS: A total of 105 genetics APPs (93%) completed the survey. Genetics APPs evaluate various patient types and populations in multiple settings, working an average of 41.3 hours and seeing 15 patients weekly. Nearly all see new (96%) and follow-up (98%) patients and utilize telemedicine (93%). Half (51%) have only worked in the genetics specialty during their career. Overall, APPs are generally satisfied with their career as a genetics APP (98%) and work-life balance (86%), and most (86%) feel they function at the top of their scope. CONCLUSION: Study findings elucidate the current state of genetics APPs. Results define the characteristics and role of an APP in the genetics specialty and will guide employers and genetics organizations to utilize APPs at the top of their scope and recruit new APPs to this exciting field. A collaborative effort is needed to increase the overall genetics workforce, decrease patient wait times, and increase access to genetics care.

MEF2C regulates NK cell effector functions through control of lipid metabolism.

Natural killer (NK) cells are a critical first line of defense against viral infection. Rare mutations in a small subset of transcription factors can result in decreased NK cell numbers and function in humans, with an associated increased susceptibility to viral infection. However, our understanding of the specific transcription factors governing mature human NK cell function is limited. Here we use a non-viral CRISPR-Cas9 knockout screen targeting genes encoding 31 transcription factors differentially expressed during human NK cell development. We identify myocyte enhancer factor 2C (MEF2C) as a master regulator of human NK cell functionality ex vivo. MEF2C-haploinsufficient patients and mice displayed defects in NK cell development and effector function, with an increased susceptibility to viral infection. Mechanistically, MEF2C was required for an interleukin (IL)-2- and IL-15-mediated increase in lipid content through regulation of sterol regulatory element-binding protein (SREBP) pathways. Supplementation with oleic acid restored MEF2C-deficient and MEF2C-haploinsufficient patient NK cell cytotoxic function. Therefore, MEF2C is a critical orchestrator of NK cell antiviral immunity by regulating SREBP-mediated lipid metabolism.

Who is most vulnerable? Factors associated with presenting to antenatal care without a male partner in Northern Tanzania.

OBJECTIVE: Male engagement in pregnancy care can be beneficial for maternal and child health outcomes. In Tanzania, pregnant women are strongly encouraged to present to their first antenatal care (ANC) appointment with a male partner, where they jointly test for HIV. For some, this presents a barrier to ANC attendance. The objectives of this study were to identify factors associated with presenting to ANC with a male partner using a cross-sectional design and to assess whether women presenting without partners had significantly delayed presentation. METHODS: Pregnant women (n = 1007) attending a first ANC appointment in Moshi, Tanzania were surveyed. Questions captured sociodemographic characteristics and measures of psychosocial constructs. RESULTS: Just over half (54%) of women presented to care with a male partner. Women were more likely to present with a male partner if they were younger than 25 years old, married, Muslim, attending ANC for their first pregnancy, and testing for HIV for the first time. Women presenting to ANC with a male partner were significantly more likely to attend ANC earlier in their pregnancy than those presenting without male partners. CONCLUSION: Policy change allowing women to present to care with other supportive family members could promote earlier presentation to first ANC. Unmarried women may be at a disadvantage in presenting to ANC when policies mandate attendance with a male partner. Male partners of multiparous women should be encouraged to provide pregnancy support even after first pregnancies, and a wholistic emphasis (beyond HIV testing) on first ANC could encourage male engagement beyond the initial appointment.

Understanding male involvement in antenatal care in the Kilimanjaro region of Tanzania: Barriers, facilitators, and opportunities for engagement.

BACKGROUND: Male engagement in antenatal care (ANC) has been recommended by the World Health Organization to improve maternal and newborn health outcomes, but implementation challenges remain. This study explored barriers, facilitators, and opportunities to improve male attendance and engagement in ANC. METHODS: In-depth interviews were conducted individually with pregnant women and male partners attending a first ANC visit at two public health facilities in Moshi, Tanzania. Interviews examined factors influencing male ANC attendance and male experiences during the clinic visit. Interviews were recorded, transcribed verbatim, and translated from Swahili into English. Transcripts were coded thematically in NVivo. MAIN FINDINGS: Constructions of masculinity both positively and negatively influenced male involvement in ANC. Individual-level barriers included a fear of HIV testing, perceptions of pregnancy as the woman's responsibility, and discomfort with ANC as a predominantly female space. Structural barriers included inability to take time off from work and long clinic wait times. The primary facilitator to male involvement was the preferential care given in the ANC clinic to women who present with a male partner. Additionally, some men desired to learn about their family's health status and felt that attending ANC was a sign of respect and love for their partner. CONCLUSIONS: Opportunities exist to improve male involvement in ANC, namely training providers to engage men beyond HIV testing and counseling. Peer programs that promote men's engagement in pregnancy could prove useful to reduce apprehension around HIV testing and dispel conceptions of ANC as only a women's healthcare space.

Psychological distress and pain related to gynecologic exams among female survivors of sexual and physical violence: A systematic review.

Gender-based violence is prevalent globally, yet the impacts of sexual and physical violence on women's experiences of routine gynecologic care are not well understood. The purpose of this systematic review of quantitative research is to describe (a) psychological distress and pain related to gynecologic exams among female survivors of sexual and physical violence and (b) differences in distress or pain between survivors and women without this history. Fourteen articles based on 12 discrete studies met the inclusion criteria. Studies were heterogeneous, with a moderate risk of bias; therefore, a descriptive summary approach was utilized rather than a meta-analytic approach. Synthesized results indicated that survivors of violence experience mild-to-severe levels of distress and mild-to-moderate levels of pain related to gynecologic exams. The findings suggest that survivors of sexual or physical violence experience higher levels of distress than women without this history (i.e., moderate to severe), and this difference was further accentuated among women with more severe posttraumatic stress symptoms (PTSS). Differences in pain by violence history and PTSS severity were not consistently observed, possibly due to a lack of variability in ratings and small sample sizes. Additional research is needed that bolsters the measurement of exam-related distress and pain, adjusts for confounding variables, and explores mechanisms by which sexual and physical violence impact care experiences. Further empirical work will be critical to developing interventions at the patient and provider levels to improve women's experiences of care.

The smallest likely pathogenic duplication of a SOX9 enhancer identified to date in a family with 46,XX testicular differences of sex development.

Copy number variants that duplicate distal upstream enhancer elements of the SOX9 gene cause 46,XX testicular differences of sex development (DSD) which is characterized by a 46,XX karyotype in an individual presenting with either ambiguous genitalia or genitalia with varying degrees of virilization, including those resembling typical male genitalia. Reported duplications in this region range in size from 24 to 780 kilobases (kb). Here we report a family with two affected individuals, the proband and his maternal uncle, harboring a 3.7 kb duplication of a SOX9 enhancer identified by clinical genome sequencing. Prior fluorescence in situ hybridization (FISH) for SRY and a multi-gene panel for ambiguous genitalia were non-diagnostic. The unaffected mother also carries this duplication, consistent with previously described incomplete penetrance. To our knowledge, this is the smallest duplication identified to-date, most of which resides in a 5.2 kb region that has been previously shown to possess enhancer activity that promotes the expression of SOX9. The duplication was confirmed by quantitative-PCR and shown to be in tandem by bidirectional Sanger sequencing breakpoint analysis. This finding highlights the importance of non-coding variant interrogation in suspected genetic disorders.

Situating reproductive coercion in the sociocultural context: An ecological model to inform research, practice, and policy in the United States.

Reproductive coercion (RC) can be conceptualized as any behavior that limits one's ability to make decisions about their reproductive health. Here, we broaden this definition to consider the impact of systemic and sociocultural factors on RC using an ecological model. Specifically, we use Bronfenbrenner's model as a framework for organizing the multilevel factors that influence reproductive coercion (RC) and its impacts on individual health. This paper is intended to offer a primer to historical, sociocultural, community, interpersonal, and individual processes that may interact to shape reproductive decision-making and its effect on individual health outcomes. We emphasize the importance of conceptualizing RC within the broader sociocultural and community context, and the potential implications for reproductive and sexual health research, clinical care, and policy in the United States.

RNA analysis of intronic variants in the LAMA2 gene detected by whole genome sequencing confirms a rare dual diagnosis of incontinentia pigmenti with limb-girdle muscular dystrophy.

We see that a multiple methods approach to diagnosis remains necessary in the era of whole genome sequencing. We also observe that reproductive risk genetic counseling can be a motivating factor for further testing along the diagnostic odyssey.

Clinical and functional characterization of germline PIK3CA variants in patients with PIK3CA-related overgrowth spectrum disorders.

Mosaic variants in the PIK3CA gene, encoding the catalytic subunit of phosphoinositide 3-kinase (PI3K), produce constitutive PI3K activation, which causes PIK3CA-related overgrowth spectrum disorders. To date, fewer than 20 patients have been described with germline alterations in PIK3CA. In this study, we describe three unrelated individuals with overgrowth and germline PIK3CA variants. These variants were discovered through whole-exome sequencing and confirmed as germline by testing multiple tissue types, when available. Functional analysis using Patient 1's fibroblast cell line and two previously reported patients' cell lines showed increased phosphorylation of AKT during cellular starvation revealing constitutive activation of the phosphoinositide-3-kinase/protein kinase B/mechanistic target of rapamycin (PI3K/AKT/mTOR) pathway. Alternatively, stimulation of the cells by fetal bovine serum produced a reduced response, indicating an activated status of the PI3K complex reducing the pathway response to further external stimulation. Additional studies utilizing Biolog Phenotype Microarray technology indicated reduced energy production when cells were exposed to growth factors stimulating the PI3K/AKT/mTOR pathway, confirming the trend observed in the AKT phosphorylation test after stimulation. Furthermore, treatment with inhibitors of the PI3K/AKT/mTOR pathway rescued the normal energy response in the patients' cells. Collectively, these data demonstrate that disease-causing germline PIK3CA variants have a functional consequence, similar to mosaic variants in the PI3K/AKT/mTOR pathway.

Expansion and mechanistic insights into de novo DEAF1 variants in DEAF1-associated neurodevelopmental disorders.

De novo deleterious and heritable biallelic mutations in the DNA binding domain (DBD) of the transcription factor deformed epidermal autoregulatory factor 1 (DEAF1) result in a phenotypic spectrum of disorders termed DEAF1-associated neurodevelopmental disorders (DAND). RNA-sequencing using hippocampal RNA from mice with conditional deletion of Deaf1 in the central nervous system indicate that loss of Deaf1 activity results in the altered expression of genes involved in neuronal function, dendritic spine maintenance, development, and activity, with reduced dendritic spines in hippocampal regions. Since DEAF1 is not a dosage-sensitive gene, we assessed the dominant negative activity of previously identified de novo variants and a heritable recessive DEAF1 variant on selected DEAF1-regulated genes in 2 different cell models. While no altered gene expression was observed in cells over-expressing the recessive heritable variant, the gene expression profiles of cells over-expressing de novo variants resulted in similar gene expression changes as observed in CRISPR-Cas9-mediated DEAF1-deleted cells. Altered expression of DEAF1-regulated genes was rescued by exogenous expression of WT-DEAF1 but not by de novo variants in cells lacking endogenous DEAF1. De novo heterozygous variants within the DBD of DEAF1 were identified in 10 individuals with a phenotypic spectrum including autism spectrum disorder, developmental delays, sleep disturbance, high pain tolerance, and mild dysmorphic features. Functional assays demonstrate these variants alter DEAF1 transcriptional activity. Taken together, this study expands the clinical phenotypic spectrum of individuals with DAND, furthers our understanding of potential roles of DEAF1 on neuronal function, and demonstrates dominant negative activity of identified de novo variants.

Mosaicism of common pathogenic MECP2 variants identified in two males with a clinical diagnosis of Rett syndrome.

Rett (RTT) syndrome, a neurodevelopmental disorder caused by pathogenic variation in the MECP2 gene, is characterized by developmental regression, loss of purposeful hand movements, stereotypic hand movements, abnormal gait, and loss of spoken language. Due to the X-linked inheritance pattern, RTT is typically limited to females. Recent studies revealed somatic mosaicism in MECP2 in male patients with RTT-like phenotypes. While detecting mosaic variation using Sanger sequencing is theoretically possible for mosaicism over ~15%-20%, several variables, including efficiency of PCR, background noise, and/or human error, contribute to a low detection rate using this technology. Mosaic variants in two males were detected by next generation sequencing (NGS; Case 1) and by Sanger re-sequencing (Case 2). Both had targeted digital PCR (dPCR) to confirm the variants. In this report, we present two males with classic RTT syndrome in whom we identified pathogenic variation in the MECP2 gene in the mosaic state (c.730C > T (p.Gln244*) in Patient 1 and c.397C > T (p.Arg133Cys) in Patient 2). In addition, estimates and measures of mosaic variant fraction were surprisingly similar between Sanger sequencing, NGS, and dPCR. The mosaic state of these variants contributed to a lengthy diagnostic odyssey for these patients. While NGS and even Sanger sequencing may be viable methods of detecting mosaic variation in DNA or RNA samples, applying targeted dPCR to supplement these sequencing technologies would provide confirmation of somatic mosaicism and mosaic fraction.

RNA analysis of the GALNS transcript reveals novel pathogenic mechanisms associated with Morquio syndrome A.

Morquio syndrome A (Mucopolysaccharidosis IVA, MPS IVA) is an autosomal recessive lysosomal storage disorder caused by deficiency of N-acetyl-galactosamine-6-sulfatase (GALNS) which catabolizes the glycosaminoglycans (GAG), keratan sulfate and chondroitin-6-sulfate. Homozygous or compound heterozygous pathogenic variants in the GALNS result in the deficiency of the enzyme and consequent GAG accumulations. DNA sequence and copy number analysis of the GALNS coding region fails to identify biallelic causative pathogenic variants in up to 15% of patients with Morquio syndrome A. RNA transcript analysis was performed to identify pathogenic alterations in two unrelated families with Morquio syndrome A in whom a single heterozygous or no pathogenic alteration was detected by standard analysis of the GALNS gene. RNA sequencing and quantitative expression analysis identified the overabundance of an aberrant GALNS transcript isoform and a reduction of the clinically relevant isoform (NM_000512.4) in the Morquio syndrome A patients from both families. The aberrant isoform (ENST00000568613.1) was produced by alternative splicing and contained intronic sequence that was likely a cryptic exon predicted to result in a reading frame shift and generation of a premature termination codon. These findings indicated that the aberrant splicing is likely the novel molecular defect in our patients. RNA transcript analysis could be useful to identify pathogenic alterations and increase the yield of molecular diagnosis in patients with Morquio syndrome A whose genetic variants are not found by standard sequencing or gene dosage analysis.

Building a multicultural peer-consultation team: Planning, implementing, and early sustainment evaluation.

This article represents an implementation-focused evaluation of a multicultural peer-consultation team situated within a psychiatry department in a large academic medical center in the Southern United States. The evaluation comprised anonymous self-report questionnaires (n = 14) as well as individual (n = 3) or group interviews (n = 10) conducted by outside independent evaluators. Participants were current and former team members (i.e., graduate trainees, mental health care providers, clinical and research staff members) who voluntarily participated in this multimethod implementation evaluation. Results indicated that attendance on the team had several important impacts on members, and most notably an increased ability to provide multiculturally competent care, that is treatment that carefully and routinely considers the influence of culture and context on patients and therefore their clinical presentation. Further, no negative impacts from participating on the team were noted. A primary strength of the team's sustainability is that participation on the team was deemed to be relevant and useful by current and former team members. A major barrier to participation on the team is competing demands, such as high clinical loads. We conclude that this model for multicultural peer-consultation holds promise as an effective and implementable educational method for mental health care professionals. We discuss strengths, limitations, and future directions for research.

Clinical findings from the landmark MEF2C-related disorders natural history study.

INTRODUCTION: MEF2C-related disorders are characterized by developmental and cognitive delay, limited language and walking, hypotonia, and seizures. A recent systematic review identified 117 patients with MEF2C-related disorders across 43 studies. Despite these reports, the disorder is not easily recognized and assessments are hampered by small sample sizes. Our objective was to gather developmental and clinical information on a large number of patients. METHODS: We developed a survey based on validated instruments and subject area experts to gather information from parents of children with this condition. No personal identifiers were collected. Surveys and data were collected via REDCap and analyzed using Excel and SAS v9.4. RESULTS: Seventy-three parents completed the survey, with 39.7% reporting a MEF2C variant and 54.8% reporting a deletion involving MEF2C. Limited speech (82.1%), seizures (86.3%), bruxism (87.7%), repetitive movements (94.5%), and high pain tolerance (79.5%) were some of the prominent features. Patients with MEF2C variants were similarly affected as those with deletions. Female subjects showed higher verbal abilities. CONCLUSION: This is the largest natural history study to date and establishes a comprehensive review of developmental and clinical features for MEF2C-related disorders. This data can help providers diagnose patients and form the basis for longitudinal or genotype-phenotype studies.

Indirect effects of dissociation on the relationship between lifetime PTSD symptoms and condomless sex among men who have sex with men with a history of childhood sexual abuse.

Posttraumatic stress disorder (PTSD) symptoms may interfere with gay, bisexual and other men who have sex with men's (MSM) ability to engage in safe sex practices. An indirect relationship with dissociation may help to elucidate the relationship between PTSD symptom severity and condomless sex among MSM with childhood sexual abuse (CSA) histories. These relationships have not previously been examined in this group, which has a unique vulnerability for HIV acquisition. A cross-sectional sample of MSM with histories of CSA (N=290) was recruited at study sites in Boston, MA, and Miami, FL. Participants had a mean age of 37.95 years (SD=11.68), 22% were African American and 29.4% identified as Latino. The sample reported a mean of 10.47 (SD=4.38) lifetime PTSD symptoms and 26.4% met the clinical threshold for dissociation. Logistic regression models (adjusted for age, education, and substance use disorder) were used to assess indirect effects of dissociation on the relationship between lifetime PTSD symptoms and condomless anal/vaginal sex episodes with serodiscordant or unknown status partners in the past 3 months. Dissociation accounted for the association between lifetime PTSD symptom severity and condomless sex episodes. The Sobel test (Sobel = 2.04, p= .042; CI 95% bias-corrected bootstrap) suggested significant indirect effects for dissociation. Dissociation among MSM with CSA histories may compromise accurate appraisals of sexual risk and safety and increase vulnerability for HIV acquisition. Further research is warranted to address HIV prevention in the context of PTSD symptom severity to improve the mental health of MSM and increase the effectiveness of HIV prevention interventions.

Male engagement guidelines in antenatal care: unintended consequences for pregnant women in Tanzania.

BACKGROUND: The meaningful engagement of male partners in antenatal care (ANC) can positively impact maternal and newborn health outcomes. The Tanzania National Plan for the Elimination of Mother to Child Transmission of HIV recommends male partners attend the first ANC appointment as a strategy for HIV prevention and treatment. This recommendation seeks to increase uptake of HIV and reproductive healthcare services, but unintended consequences of these guidelines may negatively impact women's ANC experiences. This study qualitatively examined the impact of policy promoting male engagement on women's ANC experiences. METHODS: The study was conducted in two urban clinics in Kilimanjaro Region, Tanzania. In-depth interviews were conducted with 19 participants (13 women and 6 male partners) attending a first ANC appointment. A semi-structured guide was developed, applying Kabeer's Social Relations Approach. Data were analyzed using applied thematic analysis, combining memo writing, coding, synthesis, and comparison of themes. RESULTS: Male attendance impacted the timing of women's presentation to ANC and experience during the first ANC visit. Women whose partners could not attend delayed their presentation to first ANC due to fears of being interrogated or denied care because of their partner absence. Women presenting with partners were given preferential treatment by clinic staff, and women without partners felt discriminated against. Women perceived that the clinic prioritized men's HIV testing over involvement in pregnancy care. CONCLUSIONS: Study findings indicate the need to better assess and understand the unintended impact of policies promoting male partner attendance at ANC. Although male engagement can benefit the health outcomes of mothers and newborn children, our findings demonstrate the need for improved methods of engaging men in ANC. ANC clinics should identify ways to make clinic settings more male friendly, utilize male attendance as an opportunity to educate and engage men in pregnancy and newborn care. At the same time, clinic policies should be cognizant to not discriminate against women presenting without a partner.

Preimplantation Genetic Testing for Kidney Disease-Related Genes: A Laboratory's Experience.

INTRODUCTION: Recent literature highlights the clinical utility of genetic testing for patients with kidney disease. Genetic testing provides significant benefits for reproductive risk counseling, including the option of in vitro fertilization with preimplantation genetic testing for monogenic disease (PGT-M). PGT-M allows for a significant reduction in risk for a pregnancy affected with the familial disease. We aim to summarize our experience with PGT-M for genes with kidney involvement as either a primary or secondary feature of the disease. METHODS: All PGT-M tests performed by the reference laboratory between September 2010 and July 2020 were reviewed for clinical indication and cases for which the disease tested included a renal component. Each patient referred for PGT-M had an existing molecular genetic diagnosis themselves or in their family. Frequency of each condition, gene, inheritance pattern, and year over year increase in referral cases was analyzed. RESULTS: In the study cohort, the most common disease targeted was autosomal dominant polycystic kidney disease, caused by pathogenic variants in the PKD1 or PKD2 genes, which accounted for 16.5% (64/389) of cases. The 5 most common referral indications accounted for 51.9% (202/389) of the cases. Autosomal recessive inheritance accounted for 52.0% (26/50) of conditions for which PGT-M was performed. The number of PGT-M tests performed for conditions that included either primary or secondary kidney disease increased from 5 cases in 2010 to 47 cases in the 2020 study period. DISCUSSION/CONCLUSION: These data suggest that the pursuit of PGT-M by couples at risk for passing on conditions with a kidney component is common and has significantly increased since 2010. With this rising trend of patients undergoing PGT-M and the prerequisite of molecular genetic confirmation in the PGT-M process, this study underscores the importance of the reproductive component to a molecular genetic diagnosis for patients with kidney disease, especially as the accessibility of genetic testing and utilization by nephrologists grows.

Attitudes Toward Pregnancy Among Women Enrolled in Prevention of Mother-to-Child Transmission of HIV (PMTCT) Services in Moshi, Tanzania.

For pregnant women living with HIV (WLWH), feelings about pregnancy may influence their emotional well-being and health seeking behaviors. This study examined attitudes toward pregnancy and associated factors among women enrolled in prevention of mother-to-child transmission of HIV (PMTCT) services in Moshi, Tanzania. 200 pregnant WLWH were enrolled during their second or third trimester of pregnancy and completed a structured survey. Univariable and multivariable regression models examined factors associated with attitudes toward pregnancy, including demographics, interpersonal factors, and emotional well-being. Attitudes toward the current pregnancy were generally positive, with 87% of participants reporting feeling happy about being pregnant. In the final multivariable model, having higher levels of partner support, being newly diagnosed with HIV, and having fewer children were significantly associated with more positive attitudes toward their pregnancy. Findings point to a need for tailored psychosocial support services in PMTCT, as well as comprehensive reproductive health care for WLWH.

Comprehensive investigation of the phenotype of MEF2C-related disorders in human patients: A systematic review.

MEF2C-related disorders (aka MEF2C-haploinsufficiency) are caused by variations in or involving the MEF2C gene and are characterized by intellectual disability, developmental delay, lack of speech, limited walking, and seizures. Despite these findings, the disorder is not easily recognized clinically. We performed a systematic review following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines to assemble the most comprehensive list of patients and their phenotypes. Through searching PubMed, Web of Science, and MEDLINE, 43 articles met the inclusion criteria and were fully reviewed. One hundred and seventeen patients were identified from these publications with most having a phenotype of intellectual disability, developmental delay, seizures, hypotonia, absent speech, inability to walk, stereotypic movements, and MRI abnormalities. Nonclassical findings included one patient with a question mark ear, two patients with a jugular pit, one patient with a unique neuroendocrine finding, and nine patients that did not have MEF2C deletions or disruptions but may be affected due to a positional effect on MEF2C. This systematic review characterizes the phenotype of MEF2C-related disorders, documents the severity of this condition, and will help providers to better diagnose and care for patients and their families. Additionally, this compiled information provides a comprehensive resource for investigators interested in pursuing specific genotype-phenotype correlations.