Search

Background: Joubert syndrome (JS) is a genetically heterogeneous disorder; its genetic etiology involves more than 35 genes, and a limited number of studies have investigated the pathogenic mechanism of variants in patients with JS. RNA splicing analysis is critical to determine the functional significance for noncanonical splicing variants. Methods: Whole exome sequencing was performed to screen the causative gene variants in a JS family. Sanger sequencing was used to verify the variants. cDNA PCR products were analyzed and functional experiments were performed to determine the pathogenicity of the variants. Results: The clinical phenotypes and CPLANE1 variants in the JS patient were analyzed and proved consistent. We identified two novel heterozygous variants of CPLANE1 in the proband first, including c.4459del (frameshift variant) and c.7534‐14G > A (intronic variant). We analyzed the pathogenic consequences of the 2 variants and classified the c.4459del as likely pathogenic according to the ACMG/AMP guidelines; however, the pathogenic significance of c.7534‐14G > A was uncertain. Furthermore, we performed RNA splicing analysis and revealed that the noncanonical splicing variant (c.7534‐14G > A) caused aberrant exon 37 skipping. It produced an aberrant transcript that was predicted to encode a C‐terminal truncated protein. Conclusions: The genetic variation spectrum of JS caused by CPLANE1 was updated. Two novel variants further deepened our insight into the disease's molecular mechanism and confirmed the significance of diagnostic whole‐exome sequencing. [ABSTRACT FROM AUTHOR]

Study Question: Can whole genome sequencing (WGS) offer a relatively cost-effective approach for embryonic genome-wide haplotyping and preimplantation genetic testing (PGT) for monogenic disorders (PGT-M), aneuploidy (PGT-A) and structural rearrangements (PGT-SR)?Summary Answer: Reliable genome-wide haplotyping, PGT-M, PGT-A and PGT-SR could be performed by WGS with 10× depth of parental and 4× depth of embryonic sequencing data.What Is Known Already: Reduced representation genome sequencing with a genome-wide next-generation sequencing haplarithmisis-based solution has been verified as a generic approach for automated haplotyping and comprehensive PGT. Several low-depth massively parallel sequencing (MPS)-based methods for haplotyping and comprehensive PGT have been developed. However, an additional family member, such as a sibling, or a proband, is required for PGT-M haplotyping using low-depth MPS methods.Study Design, Size, Duration: In this study, 10 families that had undergone traditional IVF-PGT and 53 embryos, including 13 embryos from two PGT-SR families and 40 embryos from eight PGT-M families, were included to evaluate a WGS-based method. There were 24 blastomeres and 29 blastocysts in total. All embryos were used for PGT-A. Karyomapping validated the WGS results. Clinical outcomes of the 10 families were evaluated.Participants/materials, Setting, Methods: A blastomere or a few trophectoderm cells from the blastocyst were biopsied, and multiple displacement amplification (MDA) was performed. MDA DNA and bulk DNA of family members were used for library construction. Libraries were sequenced, and data analysis, including haplotype inheritance deduction for PGT-M and PGT-SR and read-count analysis for PGT-A, was performed using an in-house pipeline. Haplotyping with a proband and parent-only haplotyping without additional family members were performed to assess the WGS methodology. Concordance analysis between the WGS results and traditional PGT methods was performed.Main Results and the Role Of Chance: For the 40 PGT-M and 53 PGT-A embryos, 100% concordance between the WGS and single-nucleotide polymorphism (SNP)-array results was observed, regardless of whether additional family members or a proband was included for PGT-M haplotyping. For the 13 embryos from the two PGT-SR families, the embryonic balanced translocation was detected and 100% concordance between WGS and MicroSeq with PCR-seq was demonstrated.Limitations, Reasons For Caution: The number of samples in this study was limited. In some cases, the reference embryo for PGT-M or PGT-SR parent-only haplotyping was not available owing to failed direct genotyping.Wider Implications Of the Findings: WGS-based PGT-A, PGT-M and PGT-SR offered a comprehensive PGT approach for haplotyping without the requirement for additional family members. It provided an improved complementary method to PGT methodologies, such as low-depth MPS- and SNP array-based methods.Study Funding/competing Interest(s): This research was supported by the research grant from the National Key R&D Program of China (2018YFC0910201 and 2018YFC1004900), the Guangdong province science and technology project of China (2019B020226001), the Shenzhen Birth Defect Screening Project Lab (JZF No. [2016] 750) and the Shenzhen Municipal Government of China (JCYJ20170412152854656). This work was also supported by the National Natural Science Foundation of China (81771638, 81901495 and 81971344), the National Key R&D Program of China (2018YFC1004901 and 2016YFC0905103), the Shanghai Sailing Program (18YF1424800), the Shanghai Municipal Commission of Science and Technology Program (15411964000) and the Shanghai 'Rising Stars of Medical Talent' Youth Development Program Clinical Laboratory Practitioners Program (201972). The authors declare no competing interests.Trial Registration Number: N/A. [ABSTRACT FROM AUTHOR]

The interaction between the immune cells and the host immune system with the tumor cells is significantly associated with the initiation and progression of prostate adenocarcinoma (PRAD), whereas the application of immune-related genes (IRGs) for the prognosis evaluation of PRAD patients is still lacking. In this study, we aimed to identify IRGs with prognostic values and to develop a clinically effective risk model. Wilcoxon rank-sum test and univariate Cox analysis were applied to identify the differentially expressed immune-related genes (DEIRGs) related to the survival of PRAD patients. The Least absolute shrinkage and selection operator (LASSO) analysis was performed to identify the independent prognostic DEIRGs and to establish an immune risk score prognostic model. The reliability and veracity of the prognostic model were validated in PRAD patients from the internal cohort (The Cancer Genome Atlas, TCGA dataset) and the external cohort (International Cancer Genome Consortium, ICGC dataset), respectively. Six of the 193 identified DEIRGs were survival-associated in PRAD patients. Five prognostic DEIRGs ( SLPI, NOX1, DES, BIRC5 and AMH ) were selected to construct the immune-related prognostic model with optimal robustness. In the 2 independent cohorts we chose, PRAD patients could be effectively stratified according to our risk model. Patients with high risk scores had worse survival. Clinical correlation analysis proved that the risk score was associated with advanced clinicopathologic features. Multivariate analysis indicated that the risk model was an independent prognostic indicator. We also established a nomogram based on the risk score model for clinical application. Additionally, the risk score model was correlated with immune cell infiltration and reflected the status of the immune microenvironment. The prognostic value of the five immune-related genes used in the prognostic model was also validated. Our immune-related prognostic model was an effective tool that could not only serve as a predictor for prognosis, but also provide potential prognostic and therapeutic molecular biomarkers for optimizing personalized therapies in clinical practice., Competing Interests: None., (AJCR Copyright © 2022.)

Existing staging system for prognosis evaluating for Skin Cutaneous Melanoma (SKCM) patients had defects of subjective, inaccuracy and inconsistently, therefore, to identify specific and applicable prognostic markers and promote personalized therapeutic interventions is urgently required. This study aims to build a robust autophagy-related genes (ARGs) signature for prognosis monitoring of SKCM patients. We determined 26 ARGs as differentially expressed autophagy-related genes (DEARGs) from 103 SKCM and 23 normal skin samples in GSE15605 and GSE3189 datasets. Optimal prognostic DEARGs composed the risk model were screened and verified in 458 SKCM patients in TCGA cohort as the training cohort and 209 patients in GSE65904 as the test cohort. Finally, 4 optimal independent prognostic DEARGs ( CAPNS1, DAPK2, PARP1 and PTK6 ) were filtered out in the training cohort to establish the risk model. A prognostic nomogram was established for quantitative survival prediction. The risk model grouped high-risk SKCM cancer patients exhibited significantly shorter survival times in both training and test cohorts. The area under the ROC curve for risk score model was 0.788 and 0.627 in the training and test cohorts indicated the risk model was relatively accurate for prognosis monitoring. Clinical correlation analysis exhibited that risk score was an independent predictor for prognosis significantly associated with T/N classification. The prognostic value of the 4 risk genes formed the risk model was also validated respectively. We identified a novel autophagy-related signature for prognosis monitoring. It has the potential to be an independent prognostic indicator and can benefit targeted therapy., Competing Interests: None., (AJCR Copyright © 2021.)