Congress of Neurological Surgeons Systematic Review and Evidence-based Guidelines Update on the Role of Neuropathology in the Management of Progressive Glioblastoma in Adults

Target population These recommendations apply to adult patients with progressive or recurrent glioblastoma (GBM).QuestionFor adult patients with progressive glioblastoma does testing for Isocitrate Dehydrogenase (IDH) 1 or 2 mutations provide new additional management or prognostic information beyond that derived from the tumor at initial presentation?RecommendationLevel III: Repeat IDH mutation testing is not necessary if the tumor is histologically similar to the primary tumor and the patient’s clinical course is as expected. Question For adult patients with progressive glioblastoma does repeat testing for MGMT promoter methylation provide new or additional management or prognostic information beyond that derived from the tumor at initial presentation and what methods of detection are optimal?Recommendation Level III: Repeat MGMT promoter methylation is not recommended. Question For adult patients with progressive glioblastoma does EGFR amplification or mutation testing provide management or prognostic information beyond that provided by histologic analysis and if performed on previous tissue samples, does it need to be repeated?RecommendationLevel III: In cases that are difficult to classify as glioblastoma on histologic features EGFR amplification testing may help in classification. If a previous EGFR amplification was detected, repeat testing is not necessary. Repeat EGFR amplification or mutational testing may be recommended in patients in which target therapy is being considered.Question For adult patients with progressive glioblastoma does whole genome or large panel sequencing provide management or prognostic information beyond that derived from histologic analysis?RecommendationLevel III: Primary or repeat whole genome or large panel sequencing may be considered in patients who are eligible or interested in molecularly guided therapy or clinical trials.QuestionFor adult patients with progressive glioblastoma should immune checkpoint biomarker testing be performed to provide management and prognostic information beyond that obtained from histologic analysis?RecommendationLevel III: The current evidence does not support making PD-L1 or mismatch repair (MMR) enzyme activity a component of standard testing.QuestionFor adult patients with progressive glioblastoma are there meaningful biomarkers for bevacizumab responsiveness and does their assessment provide additional information for tumor management and prognosis beyond that learned by standard histologic analysis?RecommendationLevel III: No established Bevacizumab biomarkers are currently available based upon the inclusion criteria of this guideline.

Comparison between Fluorimetry (Qubit) and Spectrophotometry (NanoDrop) in the Quantification of DNA and RNA Extracted from Frozen and FFPE Tissues from Lung Cancer Patients: A Real-World Use of Genomic Tests

Background and Objectives: Panel-based next-generation sequencing (NGS) has been carried out in daily clinical settings for the diagnosis and treatment guidance of patients with non-small cell lung cancer (NSCLC). The success of genomic tests including NGS depends in large part on preparing better-quality DNA or RNA; however, there are no established operating methods for preparing genomic DNA and RNA samples. Materials and Methods: We compared the following two quantitative methods, the QubitTM and NanoDropTM, using 585 surgical specimens, 278 biopsy specimens, and 82 cell block specimens of lung cancer that were used for genetic tests, including NGS. We analyzed the success rate of the genomic tests, including NGS, which were performed with DNA and RNA with concentrations that were outliers for the Qubit Fluorometer. Results: The absolute value for DNA concentrations had a tendency to be higher when measured with NanoDropTM regardless of the type of specimen; however, this was not the case for RNA. The success rate of DNA-based genomic tests using specimens with a concentration below the lower limit of QubitTM detection was as high as approximately 96%. At less than 60%, the success rate of RNA-based genomic tests, including RT-PCR, was not as satisfactory. The success rates of the AmpliSeqTM DNA panel sequencing and RNA panel sequencing were 77.8% and 91.5%, respectively. If at least one PCR amplification product could be obtained, then all RNA-based sequencing was performed successfully. Conclusions: The concentration measurements with NanoDropTM are reliable. The success rate of NGS with samples at concentrations below the limit of detection of QubitTM was relatively higher than expected, and it is worth performing PCR-based panel sequencing, especially in cases where re-biopsy cannot be performed.

Gene Panel Sequencing of Patients With Monogenic Diabetes Brings to Light Genes Typically Associated With Syndromic Presentations

Gene panel sequencing (NGS) offers the possibility to analyze rare forms of monogenic diabetes (MgD). To that end, 18 genes were analyzed in 1676 patients referred for MODY genetic testing. <p>Among the 307 patients with a molecular diagnosis of MgD, 55 (17.9%) were mutated in a gene associated with a genetic syndrome. Eight percent (n=25) of the patients with mutations carried the m.3243A>G variant associated with MIDD (Maternally inherited diabetes and deafness). At time of referral very little had reported hearing loss or any other element of the typical syndromic presentation. Six percent of the patients were mutated in <i>HNF1B</i> even though the typical extra-pancreatic features were not known at time of referral. Surprisingly the third most prominent etiology in these rare forms was the <i>WFS1</i> gene accounting for 2.9% of the patients with pathogenic mutations (n=9). None of them depicted a Wolfram syndrome presentation even though some features were reported in 6/9 patients. </p> <p>Restricting the analysis of certain genes to patients with the respective specific phenotypes would miss out those with partial presentations. These results therefore underlie the undisputable benefit of NGS strategies even though the situation implies cascade consequences both for the molecular biologist and the clinician.</p>

Gene Panel Sequencing of Patients With Monogenic Diabetes Brings to Light Genes Typically Associated With Syndromic Presentations

Gene panel sequencing (NGS) offers the possibility to analyze rare forms of monogenic diabetes (MgD). To that end, 18 genes were analyzed in 1676 patients referred for MODY genetic testing. <p>Among the 307 patients with a molecular diagnosis of MgD, 55 (17.9%) were mutated in a gene associated with a genetic syndrome. Eight percent (n=25) of the patients with mutations carried the m.3243A>G variant associated with MIDD (Maternally inherited diabetes and deafness). At time of referral very little had reported hearing loss or any other element of the typical syndromic presentation. Six percent of the patients were mutated in <i>HNF1B</i> even though the typical extra-pancreatic features were not known at time of referral. Surprisingly the third most prominent etiology in these rare forms was the <i>WFS1</i> gene accounting for 2.9% of the patients with pathogenic mutations (n=9). None of them depicted a Wolfram syndrome presentation even though some features were reported in 6/9 patients. </p> <p>Restricting the analysis of certain genes to patients with the respective specific phenotypes would miss out those with partial presentations. These results therefore underlie the undisputable benefit of NGS strategies even though the situation implies cascade consequences both for the molecular biologist and the clinician.</p>

Identification of Genetic Mutations in TYR and OCA2 Genes in Congenital Families with Oculocutaneous Albinism (OCA)

Aim: The objective of the present study was to recruit congenital families of oculocutaneous albinism (OCA) and mutations in TYR and OCA2 genes are identified, which is further expanding the mutation spectrum in this population. Methods: Two consanguineous families with OCA were recruited and whole blood was collected. Clinical examination was carried out to determine the visual acuity and related eye, skin and hair examinations. Genomic DNA was extracted by standard phenol-chloroform method. Targeted exome sequencing by TruSight one sequencing panel sequencing was carried out. Sanger sequencing was performed for mutation detection in tyrosinase (TYR) and the OCA2 genes and co-segregation in OCA families. Results: Clinically, the affected individuals of two OCA families showed clinical characteristics including white to pale skin, white or blonde hairs, irritant to light, nystagmus and reduced vision. DNA sequencing showed the genetic mutation of TYR and OCA2 genes in two OCA families. In family 1, the nucleotide variant (c.1255G>A; p.Gly419Arg) was detected inTYR gene, while in another family, the splice-site variant c.1045-15T>G was identified in OCA2. Conclusion: This study concluded that identification of TYR and OCA2 mutations in OCA disease are commonly associated with the population where the consanguinity is persistent. These findings expanded the molecular basis of oculocutaneous albinism in Pakistani families and established the mode of genetic counselling and for diagnostic outcome. Keywords: Consanguineous families; Oculocutaneous albinism (OCA); mutations; tyrosinase (TYR); OCA2 gene.