Asynchronous consult report generation for pharmacogenomic clinical support: Time and motion.

As pharmacogenomic (PGx) testing becomes more commonplace in clinical practice, appropriate application of laboratory data to all relevant medications becomes necessary to maximize PGx value. However, many clinicians lack PGx knowledge and confidence, so prescribers may appreciate clinical support when applying PGx data to a patient's entire medication list. Pharmacists routinely provide PGx consult support, and asynchronous written consults may improve logistical simplicity, but specific process steps and time expectations are less settled. Four pharmacists produced written consult reports for 18 patient cases across three rounds of review. Discussion took place before each of the three rounds to drive consensus in steps, process, and resources used. Time per process step was tracked in the third round. Asynchronous written PGx consult reports generally required less than 30 min to generate if no more than 2 medications had PGx-based guidance, but that time more than doubled when more medications require PGx-based guidance. After three rounds of review, pharmacists found consensus regarding an optimal workflow for generating a PGx consult. Findings from this study may support pharmacist training, practice management, and expectation management for asynchronous written PGx consult development.

COVID-19 host genetic risk study conducted at community pharmacies: Implications for public health, research and pharmacists' scope of practice.

Community pharmacists serve a large, diverse population of patients, resulting in the potential to utilize community pharmacies as recruitment sites for clinical research. Beyond traditional roles as one of the most accessible health care professionals in the US healthcare system, pharmacists have played a major role in the response to the COVID-19 pandemic, administering hundreds of thousands of vaccines and tests. However, less emphasis is placed on the ability to leverage community pharmacies as research-focused partners for clinical studies. In this study, we demonstrate the feasibility and workflow of recruiting study participants from community pharmacies and confirm genetic markers of COVID-19 susceptibility. Specific genetic markers include those associated with COVID-19 infection risk (ACE2, TMEM27, and RAVER1), difficulty breathing (NOTCH4), and hospitalization (OAS3). In addition, collaboration with a clinical laboratory allowed for a more seamless consenting process without substantial training needs or workflow disruption at the community pharmacy site. The COVID-19 pandemic has demonstrated that the expansion of pharmacists' scope of practice is a key factor in managing the population health crisis; this study demonstrates that pharmacies can also advance clinical research studies by serving as sites for patient recruitment from a large, diverse, and ambulatory study population.

Reproducible method for assessing the effects of blue light using in vitro human skin tissues.

INTRODUCTION: High-intensity visible light (HEV), also referred to as blue light, has a wavelength of 400-500 nm and accounts for approximately one-third of the visible light. Blue light is also emitted from electronic devices and artificial indoor lighting. Studies have shown that exposure of human skin cells to light emitted from electronic devices, even as short as 1 h, can cause an increase in reactive oxygen species (ROS), apoptosis and necrosis. Despite comprising a significant portion of the light spectrum, the effects of HEV light have not been studied as extensively. This is in part due to a lack of suitable in vitro testing methods. This work was conducted in order to develop a reproducible testing method for assessing the effects of blue light on the skin. METHODS: Testing was performed using a full thickness, 3D in vitro skin tissue model. Different exposure protocols were tested to (1) determine the biological effects of blue light on the skin and (2) to identify an appropriate exposure for routine testing of cosmetic materials that may protect the skin from blue light damage. Gene expression and protein biomarkers were measured using qPCR, ELISA and immunohistochemical (IHC) methods. RESULTS: Our work demonstrates that daily exposure to blue light produced dose-and-time-dependent changes in biomarkers associated with skin damage. Exposure to blue light for 6 h for 5 consecutive days (total intensity of 30 J/cm2 ) increased the expression of genes that regulate inflammation and oxidative stress pathways and decreased the expression of genes that maintain skin barrier and tissue integrity. Exposure to blue light significantly increased protein biomarkers associated with ageing, inflammation and tissue damage. IHC staining confirmed changes in collagen, filaggrin and NQO1 protein expression. Treatment with ascorbic acid inhibited the effects of blue light, demonstrating a role in protection from blue light. CONCLUSION: Our results showed that consistent blue light exposure produced skin damage via alterations in biological pathways that are associated with skin ageing. This work provides a new, reproducible in vitro testing method for assessing the effects of blue light on human skin using gene expression, protein ELISA and IHC staining.

INTRODUCTION: La lumière visible à haute énergie (VHE), également appelée lumière bleue, a une longueur d'onde de 400 à 500 nm et représente environ un tiers de la lumière visible. La lumière bleue est également émise par les appareils électroniques et l'éclairage intérieur artificiel. Des études ont montré que l'exposition des cellules cutanées humaines à la lumière émise par les appareils électroniques, même pour une période de seulement 1 h, peut entraîner une augmentation des dérivés réactifs de l'oxygène (DRO), de l'apoptose et de la nécrose. Bien qu'ils représentent une partie importante du spectre lumineux, les effets de la lumière VHE n'ont pas été étudiés aussi largement. Cela est en partie dû à un manque de méthodes de test in vitro appropriées. Ces travaux ont été réalisé afin de développer une méthode de test reproductible pour évaluer les effets de la lumière bleue sur la peau. MÉTHODES: Les tests ont été réalisés à l'aide d'un modèle de tissu cutané 3D in vitro de pleine épaisseur. Différents protocoles d'exposition ont été testés pour (1) déterminer les effets biologiques de la lumière bleue sur la peau et (2) identifier une exposition appropriée pour les tests de routine des produits cosmétiques susceptibles de protéger la peau des dommages causés par la lumière bleue. L'expression génique et les biomarqueurs protéiques ont été mesurés à l'aide des méthodes de PCR quantitative, de dosage par la méthode immuno-enzymatique ELISA et immunohistochimiques (IHC). RÉSULTATS: Nos travaux démontrent que l'exposition quotidienne à la lumière bleue a produit des modifications dépendantes de la dose et du temps dans les biomarqueurs associés aux lésions cutanées. L'exposition à la lumière bleue pendant 6 h au cours de 5 jours consécutifs (intensité totale de 30 J/cm2) a augmenté l'expression des gènes qui régulent l'inflammation et les voies du stress oxydatif, et a diminué l'expression des gènes qui maintiennent la barrière cutanée et l'intégrité tissulaire. L'exposition à la lumière bleue a significativement augmenté les biomarqueurs protéiques associés au vieillissement, à l'inflammation et aux lésions tissulaires. La coloration par IHC a confirmé les modifications de l'expression du collagène, de la filaggrine et de la protéine NQO1. Le traitement par acide ascorbique a inhibé les effets de la lumière bleue, démontrant un rôle dans la protection contre la lumière bleue. CONCLUSION: Nos résultats ont montré qu'une exposition continue à la lumière bleue produisait des lésions cutanées par le biais d'altérations des voies biologiques associées au vieillissement de la peau. Ces travaux fournissent une nouvelle méthode de test in vitro reproductible pour évaluer les effets de la lumière bleue sur la peau humaine à l'aide de l'expression des gènes, du test ELISA de détection de protéines et de la coloration IHC.

Identification of a sex-stratified genetic algorithm for opioid addiction risk.

The opioid epidemic has had a devastating impact on our country, with wide-ranging effects on healthcare, corrections, employment, and social systems. Programs have been put in place for monitoring prescriptions, initiating and expanding medications for opioid use disorder, and harm reduction (i.e., naloxone distribution, needle exchanges). However, opportunities for personalization of opioid therapy based on addiction risk have been limited. The goal of the present study was to develop an objective risk assessment algorithm based on genetic markers that are correlated with opioid use disorder (OUD). A total of 180 single-nucleotide polymorphisms (SNPs) were tested in patients with and without OUD. SNPs selected for testing were associated with opioid metabolism and drug reward pathways based on previous studies. Of the 394 patients recruited, 200 had OUD and 194 served as controls without OUD but with prior opioid exposure. Logistic regression analyses stratified by sex identified ten unique SNPs in females and nine unique SNPs in males that were significantly associated with OUD. A Genetics Opioid Risk Score (GenORs) was calculated by counting the number of OUD risk-associated SNPs/genotypes for each patient. To evaluate the discrimination of the GenORs, a receiver operating characteristic (ROC) curve for each sex was generated and determined to be sensitive and specific. This represents the first published example of a sex-based genetic risk score with potential to predict OUD, and the first OUD algorithm to include opioid-associated pharmacokinetic genes.

Pharmacist Consult Reports to Support Pharmacogenomics Report Interpretation.

BACKGROUND: The clinical implementation of pharmacogenomics (PGx) has often involved teams that include pharmacists. PGx laboratories often provide baseline information within the laboratory report that is based on Food and Drug Administration and Clinical Pharmacogenomics Implementation Consortium guidance, but information is often provided independent of concurrent disease states or medication use, among other clinical factors. Major challenges to widescale implementation of PGx include lack of physician experience or confidence in interpreting the data. The purpose of this paper is to describe how pharmacists can help further personalize PGx information and identify clinical recommendations for a given patient. METHODS: This work was performed as a secondary objective of a study evaluating genetic biomarkers of opioid addiction risk. This portion of the study utilized a descriptive analysis of pharmacist consult reports that consist of individualized, patient-level clinical recommendations that take into account current medications, current health conditions, and PGx data. A panel of 60 common PGx targets were tested among patients being treated for chronic pain or opioid use disorder (OUD). A pharmacist consult report was generated and compared with standard laboratory reporting of general PGx information. RESULTS: Of the 252 patients, PGx reports for 198 (78.6%) contained red and/or yellow clinical decision support flags for medications with actionable or informative PGx guidance for currently prescribed medications. Pharmacists recommended modifications to current prescriptions for 31 (53%) of the patients with actionable flags and 17 (12%) of the patients with informative flags. Drug classes most commonly included medications for cardiology, depression and anxiety, pain (opioids) and gastrointestinal management. Taken together, 24.2% of the actionable and informative flags had immediate clinical value based on the pharmacist's review. An additional 217 (86%) received one or more clinical recommendations not related to PGx. CONCLUSION: While PGx provides another opportunity for pharmacotherapy personalization, PGx data must be considered within the context of other patient-specific factors. Pharmacists were able to streamline the PGx report flags and identify other pharmacotherapy interventions following application of patient-specific data, thereby developing a brief report of recommendations for the patient's prescriber(s). Engaging clinical pharmacists in the PGx clinical decision process may help to facilitate more widespread PGx implementation.

Identification of a novel gene set in human cumulus cells predictive of an oocyte's pregnancy potential.

OBJECTIVE: To identify a gene expression signature in human cumulus cells (CCs) predictive of pregnancy outcome across multiple clinics, taking into account the clinic and patient variations inherent in IVF practice. DESIGN: Retrospective analysis of single human cumulus-oocyte complexes with the use of a combined microarray and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) approach. SETTING: Multiple private IVF clinics. PATIENT(S): Fifty-eight patients. Samples from 55 patients underwent qRT-PCR analysis, and samples from 27 patients resulted in live birth. INTERVENTION(S): Gene expression analysis for correlation with pregnancy outcome on individual human CCs collected immediately after oocyte retrieval. Pregnancy prediction analysis used leave-one-out cross-validation with weighted voting. MAIN OUTCOME MEASURE(S): Combinatorial expression of 12 genes in 101 samples from 58 patients. RESULT(S): We found a set of 12 genes predictive of pregnancy outcome based on their expression levels in CCs. This pregnancy prediction model had an accuracy of 78%, a sensitivity of 72%, a specificity of 84%, a positive predictive value of 81%, and a negative predictive value of 76%. Receiver operating characteristic analysis found an area under the curve of 0.763 ± 0.079, significantly greater than 0.5 (random chance prediction). CONCLUSION(S): Gene expression analysis in human CCs should be considered in identifying oocytes with a high potential to lead to pregnancy in IVF-ET.

Chronic exposure to high levels of atrazine alters expression of genes that regulate immune and growth-related functions in developing Xenopus laevis tadpoles.

Atrazine is the most commonly detected pesticide in ground and surface waters, with seasonal spikes that often exceed the Environmental Protection Agency's "Recommended Water Quality Criterion" of 350 parts per billion (ppb). Although numerous studies have shown atrazine produces adverse effects on growth, development, immune and endocrine system functions in a wide range of species, few describe gene expression changes concurrent with atrazine-induced changes in phenotype during development. In this report, developing Xenopus laevis tadpoles were chronically exposed to 400 ppb atrazine, an environmentally relevant concentration. Affymetrix microarrays and Taqman qRT-PCR were used to define gene expression changes that underlie atrazine-induced phenotypic alterations. Atrazine significantly reduced survival and growth (weight, length and fat body size) in male and female tadpoles. Microarray analysis showed atrazine altered expression of 44 genes in male tadpoles (18 upregulated, 26 downregulated) and 77 genes in female tadpoles (23 upregulated, 54 downregulated). Classification of the genes into functional groups showed the majority of genes were associated with the following biological functions: growth and metabolism, proteolysis, fibrinogen complex formation and immune regulation. Seven genes associated with immune system function, specifically defense molecules present in the skin (e.g. magainin II, levitide A, preprocarulein, skin granule protein), were significantly downregulated in female tadpoles. These results support the idea that environmental contaminants such as atrazine compromise important gene pathways during frog development that may, ultimately, be relevant to global amphibian decline.

Gene expression changes in postmortem tissue from the rostral pons of multiple system atrophy patients.

Multiple system atrophy (MSA) is a neurodegenerative disease characterized by various degrees of Parkinsonism, cerebellar ataxia, and autonomic dysfunction. In this report, Affymetrix DNA microarrays were used to measure changes in gene expression in the rostral pons, an area that undergoes extensive damage in MSA, but not other synucleinopathies. Significant changes in expression of 254 genes (180 downregulated and 74 upregulated) occurred in pons tissue from MSA patients when compared with control patients. The downregulated genes were primarily associated with biological functions known to be impaired in Parkinson's disease (PD) and other neurological diseases; for example, downregulation occurred in genes associated with mitochondrial function, ubiquitin-proteasome function, protein modification, glycolysis/metabolism, and ion transport. On the other hand, upregulated genes were associated with transcription/RNA modification, inflammation, immune system function, and oligodendrocyte maintenance and function. Immunocytochemistry, in conjunction with quantitative image analysis, was carried out to characterize alpha-synuclein protein expression as glial cytoplasmic inclusions in the pontocerebellar tract in rostral pons tissue and to determine the relationship between the amount of aggregated alpha-synuclein protein and changes in specific gene expression. Of the regulated genes, 86 were associated with the amount of observed aggregated alpha-synuclein protein in the rostral pons tissue. These data indicate that cells in the pons of MSA patients show changes in gene expression previously associated with the substantia nigra of PD patients and/or other neurological diseases, with additional changes, for example related to oligodendrocyte function unique to MSA.

Aroclor 1254 impairs the hearing ability of Xenopus laevis.

In this study we assessed the effects of chronic, dietary exposure of Aroclor 1254 (A1254) on the hearing of Xenopus frogs. We used the auditory brainstem response (ABR) to assay changes in hearing physiology; ABR thresholds, as well as latency-intensity and amplitude-intensity profiles of the initial positive (P1) and negative (N1) peaks were measured. Two groups of animals that received 50 ppm and 100 ppm of A1254 in their diet from 5 days post-fertilization through metamorphosis were compared to a control group that received untreated chow. The results showed significant threshold elevations in the 3-4 kHz range and significantly delayed peak latencies and reduced amplitudes at these frequencies in A1254 treated animals as compared to control animals. These findings indicate that A1254 selectively damages the high-frequency sensorineural hearing system associated with the basilar papilla of frogs. This preferential damage may be related to inherent differences in the vulnerability of the basilar versus amphibian papilla in the frog. The overall results of this study are also consistent with the reported A1254-induced auditory deficits in mammals indicating that the basilar papilla of the Xenopus frog may serve as an effective model for studying the effects of A1254 on the auditory system.

Polychlorinated biphenyl exposure delays metamorphosis and alters thyroid hormone system gene expression in developing Xenopus laevis.

Polychlorinated biphenyls (PCBs) are ubiquitous environmental contaminants that disrupt thyroid hormone (TH) system function in numerous species. Previous studies have shown delayed metamorphosis in developing Xenopus laevis frogs exposed to PCBs, but the underlying molecular mechanisms have not been thoroughly investigated. In this research, developing X. laevis tadpoles were exposed to environmentally relevant concentrations (5, 50ppb) of Aroclor 1254 (A1254), a PCB mixture, dissolved in water and 0.25% dimethyl sulfoxide. Quantitative real-time reverse transcriptase polymerase chain reaction was used to measure expression of several TH system genes, other genes that regulate growth and development, and a xenobiotic response gene. Exposure to 50ppb A1254 significantly delayed metamorphosis and significantly altered gene expression of three thyroid system genes: transthyretin and types II and III deiodinase. Since all three genes regulate the amount of available, biologically active TH, PCB-induced changes in the expression of these genes may underlie alterations in metamorphic timing.