One of the most critical goals in healthcare is safe and effective drug therapy, which is directly related to an individual’s response to treatment. Precision medicine can improve drug safety in many scenarios, including polypharmacy, and it requires the development of new genetic characterization methods. In this report, we use real-time PCR, microarray techniques, and mass spectrometry (MALDI-TOF), which allows us to compare them and identify the potential benefits of technological improvements, leading to better quality medical care. These comparative studies, as part of our pharmacogenetic Five-Step Precision Medicine (5SPM) approach, reveal the superiority of mass spectrometry over the other methods analyzed and highlight the importance of updating the laboratory’s pharmacogenetic methodology to identify new variants with clinical impact.

Adverse effects can disrupt early treatment in first-episode psychosis and precipitate discontinuation. We report the use of the five-step precision medici

Journal of Medical Internet Research - International Scientific Journal for Medical Research, Information and Communication on the Internet #Preprint #PeerReviewMe: Warning: This is a unreviewed preprint. Readers are warned that the document has not been peer-reviewed by expert/patient reviewers or an academic editor, may contain misleading claims, and is likely to undergo changes before final publication, if accepted, or may have been rejected/withdrawn. Readers with interest and expertise are encouraged to sign up as peer-reviewer, if the paper is within an open peer-review period. Please cite this preprint only for review purposes or for grant applications and CVs (if you are the author). Background: Psychotic disorders, such as schizophrenia, present a significant challenge to healthcare systems due to their high disability rates and treatment costs. With discontinuation rates for antipsychotics reaching over 40% in the first year and 80% after three years, it is crucial to tailor antipsychotic selection and dosing early in treatment. Personalised precision psychiatry, underpinned by pharmacogenetics, holds considerable potential in individualising antipsychotic treatment for patients with first-episode psychosis. The internationally pioneering method called 5SPM (5-Step Precision Medicine) lets the application of pharmacogenetics to clinical practice. The recently launched PRevention and early INTervention in mental health (PRINT) programme in Salamanca, Spain, integrates this method to enhance early intervention for adolescents and young people with first-episode psychosis. Objective: The CLinical Utility of early intervention including the 5-Step Precision Medicine (5SPM) Method in first-episode Psychosis (CLUMP) project aims to explore whether an early intervention model of personalised precision psychiatry, including pharmacogenetics, improves adherence to antipsychotic medicines and, therefore, clinical and functional outcomes in young people suffering from the first episode of a psychotic illness. Methods: To achieve our objectives, we shall compare adherence to the first prescribed antipsychotic medication and clinical and functional outcomes between patients with first-episode psychosis. We shall compare two cohorts: Cohort one will receive the recently introduced PRINT programme, including the 5-Step Precision Medicine (5SPM) method and cohort two would have received the standard care provided by mental health services before the PRINT programme implementation. The primary outcome to measure treatment adherence will be all-cause discontinuation proportions during one-year follow-up. Secondary outcome measures will include pragmatic efficacy, tolerability, and functional outcome measures. For additional comparative purposes, we shall analyse environmental, clinical and pharmacogenetic information of patients with psychotic disorders of more than five years of evolution and with other mental disorders, whose data are currently stored, and ethically approved for research use. A total of 300 patients will be included in the study. Analyses will include descriptive statistics, comparison tests, Kaplan-Meier survival curves, multivariate log-rank tests, qualitative analysis, and cost-benefit evaluation, among others. Results: Ethics approval was obtained in June 2023. Recruitment for the CLUMP project began in January 2025, and enrollment for Cohort 1 will continue until May 2026. All data collection is expected to be completed by June 2027. Data analyses are estimated to take approximately six months. The project is scheduled to conclude on December, 2027. Conclusions: The CLUMP project is set to provide the first clear blueprint for implementing and evaluating the impact of personalised precision psychiatry based on pharmacogenetics in the context of early intervention programmes for the benefit of young people suffering from the first episode of a severe mental illness, such as schizophrenia. Clinical Trial: ClinicalTrials.gov NCT06453174; https://clinicaltrials.gov/study/NCT06453174

BACKGROUND: Disruption of MYBPC3 precursor mRNA splicing is a frequent genetic cause of hypertrophic cardiomyopathy (HCM). Most often, it reflects changes at canonical sites or the creation of novel splice sites. Prediction tools usually prioritize splice variants with lower efficiency when they are distant from canonical sites. These elusive variants may explain HCM in patients considered genotype-negative after DNA testing. METHODS: Massively parallel sequencing identified the previously unreported deep-intronic variant MYBPC3 c.2308+227G>A in a cohort of genotype-negative patients with HCM from Salamanca, Spain. SpliceAI predicted a benign effect (Δ score, 0.03). However, its recurrent detection prompted us to conduct extensive analyses to investigate its pathogenicity, including genetic testing, clinical assessment, family studies, splicing assays, and haplotype reconstruction. RESULTS: We identified 35 unrelated HCM probands carrying c.2308+227G>A, plus 46 relatives from 27 families, yielding 81 confirmed carriers. Of these, 56 (69%) met diagnostic criteria for HCM (66% male; mean age, 53±15 years). By age 60 years, cumulative penetrance was estimated to be 81% in women and 96% in men. The combined logarithm of the odds score of 5.51 across 20 informative families provided strong evidence of cosegregation. During follow-up of probands, no significant differences were observed in heart failure, arrhythmic events, atrial fibrillation, or mortality when compared with cohorts carrying other pathogenic MYBPC3 variants. Blood splicing assays, confirmed in cardiac tissue, showed that c.2308+227G>A disrupts splicing by the use of pre-existing cryptic donor (c.2308+299) and acceptor (c.2309−580) splice sites, generating 2 misspliced mRNAs with partial intron retention or cryptic exon inclusion, which encode truncated proteins. These cryptic sites are the leading natural, unannotated missplicing events in intron 23, and some of the most frequent across the whole MYBPC3 precursor mRNA, as revealed by SpliceVault analysis of RNA sequencing databases. This deep-intronic variant does not alter essential splice motifs but promotes the use of pre-existing cryptic sites, likely through gain of splicing regulatory enhancer elements. The identification of a shared haplotype among HCM probands carrying this variant explains its recurrent detection, consistent with an ancient founder effect. CONCLUSIONS: MYBPC3 c.2308+227G>A is a pathogenic splice-altering variant that causes HCM by amplifying natural missplicing events. These background splicing errors help explain how this elusive variant disrupts splicing without creating novel essential splice sites, thereby expanding the known mechanisms by which deep-intronic variants alter MYBPC3 splicing and ultimately contribute to HCM pathogenesis. Elusive MYBPC3 splice-altering variants should be considered in HCM patients with an unresolved genetic origin of disease.