Japanese Lung Cancer Society Guidelines for Stage IV NSCLC With EGFR Mutations

Patients with NSCLC in East Asia, including Japan, frequently contain EGFR mutations. In 2018, we published the latest full clinical practice guidelines on the basis of those provided by the Japanese Lung Cancer Society Guidelines Committee. The purpose of this study was to update those recommendations, especially for the treatment of metastatic or recurrent EGFR-mutated NSCLC. We conducted a literature search of systematic reviews of randomized controlled and nonrandomized trials published between 2018 and 2019 that multiple physicians had reviewed independently. On the basis of those studies and the advice from the Japanese Society of Lung Cancer Expert Panel, we developed updated guidelines according to the Grading of Recommendations, Assessment, Development, and Evaluation system. We also evaluated the benefits of overall and progression-free survival, end points, toxicities, and patients’ reported outcomes. For patients with NSCLC harboring EGFR-activating mutations, the use of EGFR tyrosine kinase inhibitors (EGFR TKIs), especially osimertinib, had the best recommendation as to first-line treatment. We also recommended the combination of EGFR TKI with other agents (platinum-based chemotherapy or antiangiogenic agents); however, it can lead to toxicity. In the presence of EGFR uncommon mutations, except for an exon 20 insertion, we also recommended the EGFR TKI treatment. However, we could not provide recommendations for the treatment of EGFR mutations with immune checkpoint inhibitors, including monotherapy, and its combination with cytotoxic chemotherapy, because of the limited evidence present in the literature. The 2020 Japanese Lung Cancer Society Guidelines can help community-based physicians to determine the most appropriate treatments and adequately provide medical care to their patients

Role of Site-Specific N-Glycans Expressed on GluA2 in the Regulation of Cell Surface Expression of AMPA-Type Glutamate Receptors.

The AMPA-type glutamate receptor (AMPAR), which is a tetrameric complex composed of four subunits (GluA1-4) with several combinations, mediates the majority of rapid excitatory synaptic transmissions in the nervous system. Cell surface expression levels of AMPAR modulate synaptic plasticity, which is considered one of the molecular bases for learning and memory formation. To date, a unique trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc), human natural killer-1 (HNK-1) carbohydrate, was found expressed specifically on N-linked glycans of GluA2 and regulated the cell surface expression of AMPAR and the spine maturation process. However, evidence that the HNK-1 epitope on N-glycans of GluA2 directly affects these phenomena is lacking. Moreover, it is thought that other N-glycans on GluA2 also have potential roles in the regulation of AMPAR functions. In the present study, using a series of mutants lacking potential N-glycosylation sites (N256, N370, N406, and N413) within GluA2, we demonstrated that the mutant lacking the N-glycan at N370 strongly suppressed the intracellular trafficking of GluA2 from the endoplasmic reticulum (ER) in HEK293 cells. Cell surface expression of GluA1, which is a major subunit of AMPAR in neurons, was also suppressed by co-expression of the GluA2 N370S mutant. The N370S mutant and wild-type GluA2 were co-immunoprecipitated with GluA1, suggesting that N370S was properly associated with GluA1. Moreover, we found that N413 was the main potential site of the HNK-1 epitope that promoted the interaction of GluA2 with N-cadherin, resulting in enhanced cell surface expression of GluA2. The HNK-1 epitope on N-glycan at the N413 of GluA2 was also involved in the cell surface expression of GluA1. Thus, our data suggested that site-specific N-glycans on GluA2 regulate the intracellular trafficking and cell surface expression of AMPAR

The HNK-1 epitope expressed on GluA2 enhances cell surface expression of GluA1.

<p>A cell biotinylation assay was applied to HEK293 cells expressing GluA1 with (+) or without (-) GluA2 or HNK-1-synthesizing enzymes (GlcAT-P and HNK-1ST) as indicated. Biotinylated GluA1 and GluA2 were immunoblotted with anti-GluA1 and anti-GluA2/3 polyclonal antibodies, respectively (Surface) (A). The cell lysates were also immunoblotted with the same polyclonal antibodies (Total) and an HNK-1 monoclonal antibody (B). (C) Relative intensities of surface expression levels of GluA2 or GluA1 (surface/total) were calculated and normalized with or without the HNK-1 epitope.</p

N-glycan at N370 is essential for cell surface expression of GluA2.

<p>(A) GluA2 is composed of NTD (pink), LBD (blue), transmembrane domains, and a cytoplasmic domain. NTD includes two N-glycosylation sites (N256 and N370), and N406 and N413 are located in the linker between NTD and LBD. The amino acid number was counted from the first methionine of the signal sequence. (B) A cell biotinylation assay was applied to HEK293 cells expressing GluA2 wild-type (WT) or N-glycosylation site mutants (N256S, N370S, N406S, or N413S). Biotinylated GluA2 was immunoblotted with anti-GluA2/3 polyclonal antibody (Surface). The lysates were also immunoblotted for loading control (Total). (C) HEK293 cells expressing WT or mutants were doubly immunostained. Cell surface GluA2 was stained with anti-GluA2 N-terminal monoclonal antibody (red) under nonpermeabilizing conditons. Intracellular GluA2 was subsequently stained with anti-GluA2/3 polyclonal antibody (green) after cell permeabilization.</p

The HNK-1 epitope on N-glycan at N413 enhances cell surface expression of GluA2.

<p>(A) A cell biotinylation assay was applied to HEK293 cells expressing GluA2 (WT, N256S, or N413S) with (+) or without (-) HNK-1-synthesizing enzymes (GlcAT-P and HNK-1ST). Biotinylated GluA2 was immunoblotted with anti-GluA2/3 polyclonal antibody (Surface). The cell lysates were also immunoblotted with the same polyclonal antibody (Total) and an HNK-1 monoclonal antibody. (B) Relative intensities of surface expression levels of GluA2 (surface/total) were calculated and normalized with that of WT without the HNK-1 epitope.</p

N-glycan at N370 of GluA2 regulates the intracellular trafficking of GluA1.

<p>(A) GluA1 and GluA2 (WT, N256S, N370S, or N413S) were transfected into HEK293 cells. GluA1 and GluA2 were immunoprecipitated with anti-GluA1 (right) and anti-GluA2/3 (middle) polyclonal antibodies, respectively, and then immunoblotted with each antibody. The cell lysates were also immunoblotted for loading control (Input) (left). (B) A cell biotinylation assay was applied to HEK293 cells expressing GluA1 and/or GluA2 under several conditions: GluA1 alone, GluA2WT alone, GluA1 and GluA2WT, GluA1 and GluA2N256S, or GluA1 and GluA2N370S. Biotinylated GluA1 and GluA2 were immunoblotted with anti-GluA1 and anti-GluA2/3 polyclonal antibodies, respectively (Surface). The cell lysates were also immunoblotted for loading control (Total).</p

The HNK-1 epitope is preferentially expressed on N-glycan at N413.

<p>GluA2 (WT, N256S, N370S, N406S, or N413S) was transfected into HEK293 cells with HNK-1-synthesizing enzymes (GlcAT-P and HNK-1ST). After immunoprecipitation (IP) with anti-GluA2/3 polyclonal antibody, immunoprecipitates were immunoblotted with an HNK-1 monoclonal antibody and anti-GluA2/3 polyclonal antibody (A). The cell lysates were immunoblotted with an HNK-1 monoclonal antibody (Input) (B).</p

Quantitative and Morphological Assessment of Computed Tomography-depicted Gynecomastia in Spinal and Bulbar Muscular Atrophy

令和3年

Cortical thickness of the tibial diaphysis reveals age- and sex-related characteristics between non-obese healthy young and elderly subjects depending on the tibial regions

Abstract Purpose This study aimed to evaluate the age- and sex-related characteristics in cortical thickness of the tibial diaphysis between non-obese healthy young and elderly subjects as reference data. Methods The study investigated 31 young subjects (12 men and 19 women; mean age, 25 ± 8 years) and 54 elderly subjects (29 men and 25 women; mean age, 70 ± 6 years). Three-dimensional estimated cortical thickness of the tibial diaphysis was automatically calculated for 5000–9000 measurement points using the high-resolution cortical thickness measurement from clinical computed tomography data. In 12 assessment regions created by combining three heights (proximal, central, and distal diaphysis) and four areas of the axial plane at 90° (medial, anterior, lateral, and posterior areas) in the tibial coordinate system, the standardized thickness was assessed using the tibial length. Results As structural characteristics, there were no differences in the medial and lateral thicknesses, while the anterior thickness was greater than the posterior thickness in all groups. The sex-related difference was not shown. As an age-related difference, elderly subjects showed greater or lesser cortical thickness than the young subjects, depending on the regions of the tibia. Conclusions Cortical thickness was different depending on sex, age, and regions in the tibia. The results of this study are of clinical relevance as reference points to clarify the causes of various pathological conditions for diseases. Level of evidence Level 3