Dasari, Surendra

Abstract Purpose: IgM monoclonal gammopathy of undetermined significance (MGUS) and Waldenström macroglobulinemia (WM) represent a disease spectrum with highly varied therapeutic management, ranging from observation to chemoimmunotherapy. The current classification relies solely on clinical features and does not explain the heterogeneity that exists within each of these conditions. Further investigation is warranted to shed light on the biology that may account for the clinical differences. Experimental Design: We used bone marrow (BM) clonal CD19+ and/or CD138+ sorted cells, matched BM supernatant, and peripheral blood serum from 32 patients (7 MGUS, 25 WM) to perform the first multi-omics approach including whole-exome sequencing, RNA sequencing, proteomics, metabolomics, and mass cytometry. Results: We identified three clusters with distinct pathway activation, immune content, metabolomic, and clinical features. Cluster 1 included only patients with WM and was characterized by transcriptional silencing of genes involved in cell cycle and immune response, enrichment of mitochondrial metabolism, infiltration of senescent T effector memory cells, and aggressive clinical behavior. Genetic/structural alterations of TNFAIP3 were distinct events of this cluster. Cluster 2 comprised both MGUS and WM patients with upregulation of inflammatory response, senescence and glycolysis signatures, increased activated T follicular helper and T regulatory cells, and indolent clinical behavior. Cluster 3 also included both MGUS and WM patients and exhibited intermediate features, including proliferative and inflammatory signaling, as well as glycolysis and mitochondrial metabolism. Conclusions: We have identified three distinct molecular clusters, suggesting a potential biologic classification that may have therapeutic implications.

Chronic vascular diseases affect over 25 million patients in the U.S, alone. While non-invasive therapies are available, approximately 4.5 million individuals are estimated to require a vascular bypass annually, worldwide. Autologous vascular grafts remain the standard of care; yet the absence of a suitable donor vessel results in approximately one third of patients being ineligible for autologous grafting. “Off-the-shelf” alternatives have been proposed, but have had limited pre-clinical success, primarily due to graft failure via thrombosis. Thrombotic failure risk has been shown to be dramatically reduced by the formation of a luminal quiescent endothelial cell monolayer. Recently, our group engineered an unfixed, antigen-removed (AR) extracellular matrix (ECM) scaffold from bovine saphenous vein (SV) which is minimally immunogenic and avoids in vivo thrombosis. However, the mechanism by which AR-ECM SV scaffolds prevent thrombosis remains unknown. In this study, we utilized cell culture, immunofluorescence, and RNA-seq to assess the hypothesis that hAECs seeded on the basement membrane (BM) surface of AR-ECM SV scaffolds adopt a quiescent phenotype. Cells seeded on the BM surface proliferated and underwent significantly greater XY migration than those seeded on the non-BM surface. Additionally, unlike non-BM seeded cells, BM seeding resulted in quiescent hAEC phenotype (i.e., apical polarization of podocalyxin and adherence junction protein (i.e., cadherin and β-catenin) colocalization). Finally, the transcriptional phenotype of hAECs seeded on the BM surface was similar to cells treated with simvastatin (i.e. “quiescent”) and significantly different from those treated with TNFα (i.e. “activated”) (n=6/group), as assessed by gene set enrichment and t-SNE analysis. We conclude that seeding hAECs on the BM surface of AR-SV ECM scaffolds induces a favorable “quiescent” phenotype, while an unfavorable “activated” phenotype is avoided. Ultimately, these results show that the BM surface of AR-SV-ECM scaffolds possesses inherent characteristics which promote quiescent hAEC behavior and resultant vascular homeostasis and will help inform future pre-clinical studies of this novel small diameter vascular grafting biomaterial.

Abstract Atypical meningiomas (CNS WHO grade 2) show an intermediate risk of recurrence/progression, and molecular parameters informing management following gross total resection (GTR) remain to be established. We performed comprehensive genomic analysis in 63 patients who underwent GTR of an atypical meningioma. Median age was 59.8 years, 41 patients (65.1%) were female, and 8/62 patients (12.9%) underwent radiation therapy following resection. Follow-up was available for all patients (median 10 years), with recurrence/progression in 17 (5-year PFS: 84.2%; 10-year PFS: 71.0%). Tumors were evaluated by next generation sequencing (n=61), chromosomal microarray (n=63), DNA methylation profiling (n=62), and H3K27me3 immunohistochemistry (n=62). Next generation sequencing using a CLIA-certified 50-gene Neuro-Oncology specific panel identified pathogenic alterations in NF2 (n=31; 50.8%), SMARCB1 (n=5; 8.2%), AKT1 (n=2; 3.3%), TERT (n=1, 1.6%), SMO (n=1), SUFU (n=1), TP53 (n=1), and PTEN (n=1). There was trend toward decreased PFS in patients with NF2 alterations (p=0.078) which was not statistically significant. Copy number variants (CNVs) were identified using OncoScan (ThermoFisher). The most frequent CNVs were loss -22 (n=45; 71.4%), -1p (n=30, 47.6%), -14 (n=21, 33.3%), -6q (n=18, 28.6%), -10q (n=15, 23.8%), -X (n=13, 20.6%), and -18 (n=11, 17.4%). CNVs associated with decreased PFS included -1p (p=0.015), -7p (p=0.007), -10q (p=0.0007), and -18 (p=0.0376). DNA methylation profiling was performed using the Illumina MethylationEPIC array (n=62) and evaluated using the DKFZ classifier (v12.3). Based on the highest score, tumors matched to benign (n=32; 51.6%) or intermediate (n=29; 46.7%) meningioma subclasses or showed non-contributory profiles (n=1; 1.6%). Methylation class (benign vs. intermediate) showed no association with PFS (p=0.96). H3K27 trimethylation (H3K27me3) was unequivocally lost in 4 (6.5%) tumors, which was insufficient for PFS analysis. Overall, our study supports CNV profiling of atypical meningiomas following GTR, which can be implemented using existing, clinically validated technologies. Molecular predictors could guide use of radiotherapy in these patients.

BACKGROUND A patient-derived organoid (PDO) platform may serve as a promising tool for translational cancer research. In this study, we evaluated PDO’s ability to predict clinical response to gastrointestinal (GI) cancers. METHODS We generated PDOs from primary and metastatic lesions of patients with GI cancers, including pancreatic ductal adenocarcinoma, colorectal adenocarcinoma, and cholangiocarcinoma. We compared PDO response with the observed clinical response for donor patients to the same treatments. RESULTS We report an approximately 80% concordance rate between PDO and donor tumor response. Importantly, we found a profound influence of culture media on PDO phenotype, where we showed a significant difference in response to standard-of-care chemotherapies, distinct morphologies, and transcriptomes between media within the same PDO cultures. CONCLUSION While we demonstrate a high concordance rate between donor tumor and PDO, these studies also showed the important role of culture media when using PDOs to inform treatment selection and predict response across a spectrum of GI cancers. TRIAL REGISTRATION Not applicable. FUNDING The Joan F. & Richard A. Abdoo Family Fund in Colorectal Cancer Research, GI Cancer program of the Mayo Clinic Cancer Center, Mayo Clinic SPORE in Pancreatic Cancer, Center of Individualized Medicine (Mayo Clinic), Department of Laboratory Medicine and Pathology (Mayo Clinic), Incyte Pharmaceuticals and Mayo Clinic Hepatobiliary SPORE, University of Minnesota-Mayo Clinic Partnership, and the Early Therapeutic program (Department of Oncology, Mayo Clinic).

This case report describes 2 patients with iatrogenic amyloidosis secondary to subcutaneous injections of anakinra to manage neonatal-onset multisystem inflammatory disease.