Search

Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease with respect to outcome. Features of the tumor microenvironment (TME) are associated with prognosis when assessed by gene expression profiling. However, it is uncertain whether assessment of the microenvironment can add prognostic information to the most relevant and clinically well-established molecular subgroups when analyzed by immunohistochemistry (IHC).We carried out a histopathologic analysis of biomarkers related to TME in a very large cohort (n = 455) of DLBCL treated in prospective trials and correlated with clinicopathologic and molecular data, including chromosomal rearrangements and gene expression profiles for cell-of-origin and TME.The content of PD1+, FoxP3+ and CD8+, as well as vessel density, was not associated with outcome. However, we found a low content of CD68+ macrophages to be associated with inferior progression-free survival (PFS) and overall survival (OS; P = 0.023 and 0.040, respectively) at both univariable and multivariable analyses, adjusted for the factors of the International Prognostic Index (IPI), MYC break and BCL2/MYC and BCL6/MYC double-hit status. The subgroup of PDL1+ macrophages was not associated with survival. Instead, secreted protein acidic and cysteine rich (SPARC)-positive macrophages were identified as the subtype of macrophages most associated with survival. SPARC-positive macrophages and stromal cells directly correlated with favorable PFS and OS (both, P[log rank]0.001, P[trend]0.001). The association of SPARC with prognosis was independent of the factors of the IPI, MYC double-/triple-hit status, Bcl2/c-myc double expression, cell-of-origin subtype and a recently published gene expression signature [lymphoma-associated macrophage interaction signature (LAMIS)].SPARC expression in the TME detected by a single IHC staining with fair-to-good interobserver reproducibility is a powerful prognostic parameter. Thus SPARC expression is a strong candidate for risk assessment in DLBCL in daily practice.

Erythema multiforme (EM) is an immune-mediated reaction first described by von Hebra1 in 1860, presenting typically as acrofacial target lesions. The rash may be confined to the skin (EM minor), or it may involve ocular, oral, or genital mucosa with systemic symptoms (EM major). In most cases, only 1 outbreak of EM occurs in a lifetime; however, very rarely, repeated episodes of EM may occur, known as recurrent EM.2 Few cases of pediatric recurrent EM have been reported, with the etiology being identified in less than half of these cases.3 We report a pediatric case of recurrent EM resistant to conventional treatments in which a peculiar human leukocyte antigen (HLA) haplotype has been identified.

Background Interactions between multiple myeloma plasma cells (MMPC) and the multiple myeloma (MM) niche control cell proliferation, drug resistance, and disease spreading through various adhesion molecules and chemokine receptors. Among these, junctional adhesion molecule-A (JAM-A) mediates MMPC-endothelial interactions and propagates neovascularization. Here we addressed whether JAM-A signaling mediates MMPC invasive behavior orchestrating intra- and extramedullary MM dissemination. Methods We investigated JAM-A expression with flow cytometry and immunohistochemistry in 60 MM patient bone marrow (BM) aspirates and biopsies at different disease stages with and without extramedullary disease (EMD) manifestation. We compared these results with RNA-Seq data from 647 newly diagnosed MM (NDMM) patients collected in the MMRF CoMMpass study. Using bioinformatics, we investigated JAM-A-related pathways with hierarchical cluster analysis. Subsequently, we functionally validated these JAM-A-related pathways regarding epithelial-mesenchymal transition (EMT), invasion, and MM dissemination in vitro. Results The median OS differed significantly in subjects with elevated membrane JAM-A expression levels, dividing the patients in the EMD JAM-Ahigh group with a median OS of 84.1 months, whereas median OS in JAM-Alow patients was not reached, irrespective of the EMD status (log-rank=4.19, P=.04). Immunohistochemistry analysis of BM biopsies confirmed these findings. RNA-Seq analysis corroborated the prognostic impact of JAM-A (log-rank=3.8, P=.051). High-risk patients with EMD and JAM-A expression displayed a unique gene-expression signature. Additionally, we constructed a novel expression cluster subgroup model, revealing complex molecular disease patterns and associations between clinical traits and expression markers. Strikingly, MM patients clustered by JAM-A expression levels and EMD manifestation revealed properties of epithelial-mesenchymal-transition and induced focal adhesion pathways. Notably, functional knockdown of JAM-A in MMPC in vitro not only reduced cell viability but also diminished MMPC adhesion molecules and CD138 surface expression. Furthermore, mTOR/PI3K in vitro inhibition with BEZ235 significantly downregulated surface expression in JAM-Ahigh MM cells. Interestingly, when we inhibited mTOR/PI3K in JAM-Alow MM cells, we observed JAM-A induction. Finally, ensuing functional shRNA knockdown of JAM-A halted MM invasion (P Conclusions Collectively, these data reveal JAM-A-mediated signaling critical for EMT transition and invasive behavior. Based on these findings we propose JAM-A as a promising biomarker and novel theragnostic target in MM patients.

Introduction: In this study, we report on our experience using digital pathology to overcome the severe limitations imposed on health care by the Covid-19 outbreak in Northern Italy. Social distancing had a major impact on public transportation, causing it to run with reduced timetables. This resulted in a major challenge for hospital commuters. To limit the presence in our hospital of no more than two pathologists at a time out of four, a web-based digital pathology system (DPS) was employed to work remotely. Subjects and Methods: We used a DPS in which a scanner, a laboratory information system, a storage device, and a web server were interfaced so that tissue slides could be viewed over the Internet by whole-slide imaging (WSI). After a brief internal verification test, the activity on the DPS was recorded, taking track of a set of performance and efficiency indicators. At the end of the study, 405 cases were signed out remotely. Results: Of 693 cases, 58.4% were signed out remotely by WSI, while 8.4% needed to be kept on hold to return to the original microscope slide. In three cases, at least one slide had to be rescanned. In eight cases, one slide was recut. Panel discussion by WSI was necessary in 34 cases, a condition in which all pathologists were asked for their opinion. A consultation with a more experienced colleague was necessary in 17 cases. Conclusions: We show that WSI easily allows pathologists to overcome the problems caused by the severe social distancing measures imposed by the Covid-19 pandemic. Our experience shows that soon there will not be alternatives to digital pathology, given that there is no assurance that other similar outbreaks will not occur.