In vivo import of unspliced tRNATyr containing synthetic introns of variable length into mitochondria of Leishmania tarentolae

The mitochondrial genomes of trypanosomatids lack tRNA genes. Instead, mitochondrial tRNAs are encoded and synthesized in the nucleus and are then imported into mitochondria. This also applies for tRNATyr, which in trypanosomatids contains an 11 nt intron. Previous work has defined an exon mutation which leads to accumulation of unspliced precursor tRNATyr. In this study we have used the splicing-deficient tRNATyr as a vehicle to introduce foreign sequences into the mitochondrion of Leishmania tarentolae. The naturally occurring intron was replaced by synthetic sequences of increasing length and the resulting tRNATyr precursors were expressed in transgenic cell lines. Whereas stable expression of precursor tRNAsTyr was obtained for introns up to a length of 76 nt, only precursors having introns up to 38 nt were imported into mitochondria. These results demonstrate that splicing-deficient tRNATyr can be used to introduce short synthetic sequences into mitochondria in vivo. In addition, our results show that one factor which limits the efficiency of import is the length of the molecul

An Approach to Solving the Complex Clinicogenomic Data Landscape in Precision Oncology: Learnings From the Design of WAYFIND-R, a Global Precision Oncology Registry

Oncología de precisión; Datos clinicogenómicosOncologia de precisió; Dades clinicogenòmiquesPrecision oncology; Clinicogenomic dataPrecision oncology, where patients are given therapies based on their genomic profile and disease trajectory, is rapidly evolving to become a pivotal part of cancer management, supported by regulatory approvals of biomarker-matched targeted therapies and cancer immunotherapies. However, next-generation sequencing (NGS)–based technologies have revealed an increasing number of molecular-based cancer subtypes with rare patient populations, leading to difficulties in executing/recruiting for traditional clinical trials. Therefore, approval of novel therapeutics based on traditional interventional studies may be difficult and time consuming, with delayed access to innovative therapies. Real-world data (RWD) that describe the patient journey in routine clinical practice can help elucidate the clinical utility of NGS-based genomic profiling, multidisciplinary case discussions, and targeted therapies. We describe key learnings from the setup of WAYFIND-R (NCT04529122), a first-of-its-kind global cancer registry collecting RWD from patients with solid tumors who have undergone NGS-based genomic profiling. The meaning of ‘generalizability’ and ‘high quality’ for RWD across different geographic areas was revisited, together with patient recruitment processes, and data sharing and privacy. Inspired by these learnings, WAYFIND-R’s design will help physicians discuss patient treatment plans with their colleagues, improve understanding of the impact of treatment decisions/cancer care processes on patient outcomes, and provide a platform to support the design and conduct of further clinical/epidemiologic research. WAYFIND-R demonstrates user-friendly, electronic case report forms, standardized collection of molecular tumor board-based decisions, and a dashboard providing investigators with access to local cohort-level data and the ability to interact with colleagues or search the entire registry to find rare populations. Overall, WAYFIND-R will inform on best practice for NGS-based treatment decisions by clinicians, foster global collaborations between cancer centers and enable robust conclusions regarding outcome data to be drawn, improve understanding of disparities in patients’ access to advanced diagnostics and therapies, and ultimately drive advances in precision oncology

Safety and accuracy of minimally invasive long bone fracture repair using a 2.5-mm interlocking nail: a cadaveric feline study

OBJECTIVES: The Targon Vet System (TVS) is a 2.5-mm interlocking nail that can be applied minimally invasively. The purpose of this study was to test if the TVS could be safely applied percutaneously to different feline long bones without fluoroscopic guidance. METHODS: A gap fracture was created in 96 feline humeri, femora and tibiae (n = 32/group). Paired bones were randomly assigned to two treatment groups: (1) TVS inserted percutaneously with fluoroscopy and (2) TVS inserted percutaneously without fluoroscopy. Intraoperative evaluation (complications, procedure time, attempts), radiographs (pre-/postoperative alignment, length) and anatomical dissection (neurovascular injury, rotational alignment) were compared between treatment groups. RESULTS: The use of fluoroscopy did not lead to significant differences in any of the outcome measures. Intraoperative complications predominantly occurred in the distal humerus (12/32) and the proximal femur (7/32). In total, 20/96 complications occurred with no complications for the tibia. Neurovascular structures were only damaged at the medial side of the distal humerus (10/32). CLINICAL SIGNIFICANCE: We conclude that the TVS can be safely applied percutaneously to the tibia and with limitations to the femur in normal cadaveric cats without fluoroscopy. Despite the limitations of a cadaveric study, the high number of complications is leading us to consider the humerus not safe for the TVS. A learning curve has to be expected and technical recommendations should be respected to decrease complications