Citations

As part of the Multiple Myeloma Research Foundation (MMRF) immune atlas pilot project, we compared immune cells of multiple myeloma bone marrow samples from 18 patients assessed by single-cell RNA sequencing (scRNA-seq), mass cytometry (CyTOF), and cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) to understand the concordance of measurements among single-cell techniques. Cell type abundances are relatively consistent across the three approaches, while variations are observed in T cells, macrophages, and monocytes. Concordance and correlation analysis of cell type marker gene expression across different modalities highlighted the importance of choosing cell type marker genes best suited to particular modalities. By integrating data from these three assays, we found International Staging System stage 3 patients exhibited decreased CD4+ T/CD8+ T cells ratio. Moreover, we observed upregulation of RAC2 and PSMB9, in natural killer cells of fast progressors compared with those of nonprogressors, as revealed by both scRNA-seq and CITE-seq RNA measurement. This detailed examination of the immune microenvironment in multiple myeloma using multiple single-cell technologies revealed markers associated with multiple myeloma rapid progression which will be further characterized by the full-scale immune atlas project. Significance: scRNA-seq, CyTOF, and CITE-seq are increasingly used for evaluating cellular heterogeneity. Understanding their concordances is of great interest. To date, this study is the most comprehensive examination of the measurement of the immune microenvironment in multiple myeloma using the three techniques. Moreover, we identified markers predicted to be significantly associated with multiple myeloma rapid progression.

AbstractObjectivesThe assessment of physical function is fundamental in the management of patients with idiopathic inflammatory myopathies (IIMs). We aimed to investigate the physical function of patients with IIMs compared with those with non-IIM autoimmune rheumatic diseases (AIRDs) utilizing Patient-Reported Outcome Measurement Information System (PROMIS) Physical Function (PF) data obtained in the COVAD study, an international self-reported e-survey assessing the safety of COVID-19 vaccines in AIRDs.MethodsDemographics, AIRD diagnosis, disease activity, and PROMIS PF short form-10a data were extracted from the COVAD database. PROMIS PF-10a scores were compared between disease categories and stratified by disease activity. Factors affecting PROMIS PF-10a scores other than disease activity were identified by multivariable regression analysis in patients with inactive disease.ResultsA total of 1057 IIM patients, 3635 non-IIM AIRD patients and 3981 healthy controls (HCs) responded to the COVAD e-survey from April to August 2021. Using a binomial regression model, the predicted mean of PROMIS PF-10a scores was significantly lower in IIM patients compared with non-IIM AIRD patients or HCs [36.3 (95% CI 35.5, 37.1) vs 41.3 (95% CI 40.2, 42.5) vs 46.2 (95% CI 45.8, 46.6), P < 0.001], irrespective of disease activity. The independent factors for lower PROMIS PF-10a scores in patients with inactive disease were older age, female, longer disease duration, and a diagnosis of inclusion body myositis or polymyositis.ConclusionPhysical function is significantly impaired in IIMs compared with non-IIM AIRDs or HCs, even in patients with inactive disease. Our study highlights a critical need for better strategies to minimize functional disability in patients with IIMs.

Abstract Multiple Myeloma (MM) is a hematologic malignancy marked by uncontrolled clonal expansion of plasma cells. Previous research has examined single-cell transcriptome profiles of Monoclonal gammopathy of undetermined significance (MGUS) and MM tumor microenvironment (TME) and found that natural killer (NK) cell abundance is elevated in the early stages and correlated with altered chemokine receptor expression. This study suggested the critical role of immune cells on myeloma progression from asymptomatic MGUS to symptomatic MM. Up to date, however, there are no published studies comprehensively comparing tumor and immune populations differences between MM NON-progressors (NPs) and FAST-progressors (FPs) and investigating how clonal plasma cells affect disease progression in a large cohort. Therefore, understanding how tumor and immune cells influence disease progression within symptomatic MM is of great interest.Here, we subjected CD138-negative Bone Marrow Mononuclear cells (BMMC) samples from 418 MM patients to scRNA-seq. From MMRF CoMMpass study (NCT01454297), we also have whole exome sequencing (WES) and bulk RNA-seq from CD138-positive fraction of BMMC samples. Based on time to progressive disease (TTPD), we classified patients into 2 categories. Patients with TTPD less than 18 months were classified as FAST-progressors (FPs), whereas patients with TTPD more than 5 years were classified as NON-progressors (NPs). By analyzing patient genomic alterations and its association with progression, we found that there was a significant association of slow MM progression with t(11;14) (p = 0.048), consistent with previous study. In our preliminary analysis, we profiled 83 CD138-sorted MM bone marrow samples using scRNA-seq. Interestingly, we found plasma cells from samples with the same genetic alterations tend to cluster together, highlighting the important role of genetic drivers in transcriptome profiles of plasma cells. Moreover, integrated analysis of bone marrow samples from 83 MM patients and 4 healthy donors revealed an atypical naïve-B cell subset with enrichment of cells from fast-progressors and partial expression of MS4A1. Differentially expressed genes for this naïve-B cell subset includes KLF9, BCL2L11, JOSD1, and IRS2, etc. Overall, as part of MMRF immune profiling research, this study will help to interrogate how genetic alterations and disease progression interplay MM tumor and TME and provide a sufficiently broad and valuable dataset for systematically characterizing MM at single-cell resolution. Hopefully, this study could identify novel targets for MM immunotherapies, and ultimately identify patients with high risk of fast progression for early intervention in the clinic. Citation Format: Lijun Yao, Tianjiao Wang, Kazuhiro Sato, Reyka Jayasinghe, I-Ling Chiang, Darwin D'souza, William Pilcher, Edgar Gonzalez-Kozlova, Yered Pita-Juarez, Taxiarchis Kourelis, Deon Bryant Doxie, Beena Thomas, Brian Lee, Swati Sharma Bhasin, Upadhyaya Bhaskar, Mark Fiala, Julie Fortier, Travis Dawson, John Leech, Shaji Kumar, Hearn Cho, Seunghee Kim-Schulze, Bee Raj, Stephen Oh, the MMRF Immune Profiling Research Team, John Dipersio, Ravi Vij, Adeeb Rahman, Ionnis Vlachos, Shaadi Mehr, Mark Hamilton, Daniel Auclair, Surendra Dasari, David Avigan, Madhav Dhodapkar, Sacha Gnjatic, Manoj Bhasin, Li Ding. Single-cell transcriptome profiling of multiple myeloma bone marrow samples suggests that disease progression interplays with tumor and tumor microenvironment in The MMRF CoMMpass Study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1729.

Background: Describe the incidence and associated outcomes of gastrointestinal (GI) manifestations of acute coronavirus disease 2019 (COVID-19) and multisystem inflammatory syndrome in hospitalized children (MIS-C). Methods: Retrospective review of the Viral Infection and Respiratory Illness Universal Study registry, a prospective observational, multicenter international cohort study of hospitalized children with acute COVID-19 or MIS-C from March 2020 to November 2020. The primary outcome measure was critical COVID-19 illness. Multivariable models were performed to assess for associations of GI involvement with the primary composite outcome in the entire cohort and a subpopulation of patients with MIS-C. Secondary outcomes included prolonged hospital length of stay defined as being >75th percentile and mortality. Results: Of the 789 patients, GI involvement was present in 500 (63.3%). Critical illness occurred in 392 (49.6%), and 18 (2.3%) died. Those with GI involvement were older (median age of 8 yr), and 18.2% had an underlying GI comorbidity. GI symptoms and liver derangements were more common among patients with MIS-C. In the adjusted multivariable models, acute COVID-19 was no associated with the primary or secondary outcomes. Similarly, despite the preponderance of GI involvement in patients with MIS-C, it was also not associated with the primary or secondary outcomes. Conclusions: GI involvement is common in hospitalized children with acute COVID-19 and MIS-C. GI involvement is not associated with critical illness, hospital length of stay or mortality in acute COVID-19 or MIS-C.

Abstract BackgroundAlternative splicing (AS) is a fundamental process in eukaryotes contributing to the diversity of mRNA isoforms with variable ratios of intron/exon. SF3B1 is a pivotal protein of the spliceosome machinery. Mutations in the SF3B1 gene have prognostic value in Chronic Lymphocytic Leukemia (CLL). Our previous studies have shown that SF3B1 inhibition with macrolides induces apoptosis specifically in CLL compared with normal cells. SF3B1 inhibition is associated with a widespread increase in intron retention (IR) on most transcripts, suggesting that IR can be used as a marker of spliceosome inhibition in CLL cells. However, it is unknown how the activity of SF3B1 contributes to the spliceosome activity and the poor clinical prognosis in CLL.MethodsTo better understand this process, we performed a comprehensive analysis to quantify the abundance of individual exonic and intronic mapped reads on annotated RNA-Seq transcripts derived from the B cells of 98 CLL patients and nine healthy volunteers (Normal B cells – NBC). We calculated ratios for intron and exon abundance for each transcript and use this as a measure of intron retention (IR) and a surrogate for alternative splicing (AS).ResultsWe found that 66% of CLL B-cells transcripts had significant IR elevation compared to NBC and that transcripts with high IR were associated with low expression levels and mRNA downregulation. The IR increase in CLL B-cells was independent of prognostic factors such as IgVH or SF3B1 mutations. Transcripts with the highest IR levels belonged to biological pathways associated with gene expression and RNA splicing. Also, we observed a >2-fold increase of active pSF3B1 in CLL B-cells compared to NBC. Additionally, when the CLL-B cells were treated with macrolides (pladienolide-B), a significant decrease in pSF3B1, but not total SF3B1 protein was observed. ConclusionsOur findings suggest that IR/ARS is increased in CLL and that this process is associated with SF3B1 phosphorylation and susceptibility to SF3B1 inhibitors. These data provide additional support to the relevance of AS in carcinogenesis and evidence of pSF3B1 participation in this process.

This study explores the feasibility of detecting gastroesophageal reflux disease (GERD) and Barrett’s esophagus (BE) using acoustic features extracted from speech recordings. GERD is a common condition affecting up to 27.8% of adults in the US. Chronic GERD is associated with BE, the precursor to esophageal adenocarcinoma. An automatic screening tool for GERD would improve clinical outcomes by improving early detection. Biomarkers based on acoustic features of speech may hold promise as hoarseness often co-occurs with GERD. The study cohort consisted of 49 adults with and 63 without GERD based on esophagogastroduodenoscopy or ambulatory pH studies. Speech, including sustained vowels, sentences, and reading, was systematically recorded. Voice quality was validated through auditory-perceptualratings with three speech-language pathologists. Automated, cross-validated, search across multiple machine and deep learning model types was performed using the recorded speech. Detection of condition presence and severity are explored, and model performance is reported, along with an assessment of the ability of acoustic features to discern across conditions.

INTRODUCTION: To describe the clinical, endoscopic, and histopathology features of esophageal graft-vs-host disease (GVHD). Methods: Patients with biopsy-proven esophageal GVHD diagnosed at Mayo Clinic between 2000 and 2021 were included. Results: In 43 esophageal patients, other organ GVHD was present in 58% before and 86% at esophageal GVHD diagnosis. Esophageal specific symptoms were uncommon (dysphagia 26% and odynophagia/heartburn 5%). Esophagogastroduodenoscopy was abnormal in 72% patients demonstrating erosive esophagitis, ulceration, desquamation, or rings/furrows in a diffuse or focal pattern. DISCUSSION: There should be a low threshold for esophageal biopsies for GVHD because esophageal symptoms and endoscopic findings may be nonspecific or absent.

Background and Objective Hyperdense middle cerebral artery (HDMCA) sign is an early sign seen on CT head after acute ischemic stroke indicating acute arterial thrombosis. We aimed to measure HDMCA sign objectively. Methods We conducted a retrospective analysis of prospectively collected data from 151 cases of acute ischemic stroke eligible for IV rt-PA by time criteria, admitted to Mayo Clinic from March, 2002 through June, 2011. Admission CT Head was studied to identify HDMCA sign. All patients had either 24 hour CT head or MRI head to confirm the stroke in the same vessel territory. MCA was measured in Hounsfield units and was compared to the contralateral MCA. Receiver Operating Characteristic (ROC) curve analysis was performed to calculate sensitivity and specificity. Results Mean admission NIHSS score was 11±7. HDMCA sign was seen in 93 (62%) patients. ROC curve analysis for ipsilateral MCA Hounsfield units had an area under the curve 0.88 (p<0.001, CI 0.83-0.94) which showed 50 HU or more had 48% sensitivity and 98% specificity. Ratio of ipsilateral to contralateral MCA HU had an area under the curve 0.94 (p<0.001, CI 0.90-0.98) which showed ratio of 1.23 had 67% sensitivity and 98% specificity. A combination of either ipsilateral HU more than equal to 50 or a ratio of ipsilateral to contralateral MCA HU more than equal to 1.23 had 77% sensitivity, 95% specificity, 96% positive predictive value and 72% negative predictive value. Conclusion A combination of ipsilateral HU more than equal to 50 or a ratio of ipsilateral to contralateral MCA HU more than equal to 1.23 can objectively identify HDMCA sign with good sensitivity and high specificity.

Background and Objective Hyperdense MCA (HDMCA) sign is an early sign seen on CT head after acute ischemic stroke indicating acute arterial thrombosis. It has been reported as a predictor of stroke morbidity and mortality. The relationship of size of hyperdense middle cerebral artery (MCA) sign with stroke volume and functional outcome has not been sufficiently studied. Methods We did a retrospective analysis of prospectively collected data from 206 cases of acute ischemic stroke eligible for IV rt-PA by time criteria, admitted to Mayo Clinic from March, 2002 through June, 2011. Admission CT Head was studied to identify HDMCA sign which was further subcategorized according to size. CT head at 24 hours after intravenous tissue plasminogen activator was used to calculate Alberta Stroke Program Early CT Score (ASPECTS) to measure ischemic damage area. Functional outcome was assessed using the modified Rankin score at 3 months and dichotomized into less than equal to 2 (good outcome) and more than 2 (bad outcome). Univariate analysis including t-test, Chi-square, and Fisher Exact test was used when appropriate. Results Mean admission NIHSS score was 11±7. HDMCA sign was seen in 99 (48%) patients: 100% MCA occlusion (7) 76-100% M1 (17), 50-75% M1 (17), less than 50% M1 (6), MCA dot sign (23), sylvian dot sign (21) and sylvian MCA sign, defined as any sylvian MCA occlusion bigger than a dot (8). Univariate analysis showed statistically significant associations between our HDMCA classification and ASPECTS at 24 hours (p<0.001) and modified Rankin score at 3 months (p<0.001). Smaller infarctions (ASPECTS >8) were observed more often with lower MCA clot burden; MCA dot (48%), less than 50% MCA (80%), sylvian dot (57%) and sylvian MCA (38%), 50-75% M1 (6%), 75-100% M1 (0%) and 100% MCA (0%). Good outcome (mRS≤2 at 3 months) was seen more often with lower MCA clot burden; MCA dot (52%), less than 50% MCA (67%), sylvian dot (67%) and sylvian MCA (63%). Conversely, larger HDMCA size was associated with poor outcome (mRS>2); 50-75% M1 (100%), 75-100% (88%) and 100% MCA (100%). Conclusion The size of the HDMCA sign size is an important predictor of ischemic stroke mortality and morbidity. HDMCA should be measured upon admission as it may help refine prognostication and identify patients who could benefit from endovascular rescue therapy after intravenous thrombolysis.

ABSTRACT Background and Objective Patients with dementia with Lewy bodies (DLB) may have overlapping Alzheimer's disease pathology. We investigated the longitudinal rate of tau accumulation and its association with neurodegeneration and clinical disease progression in DLB. Methods Consecutive patients with probable DLB (n = 22) from the Mayo Clinic Alzheimer's Disease Research Center and age‐matched and sex‐matched cognitively unimpaired controls (CU; n = 22) with serial magnetic resonance imaging and flortaucipir positron emission tomography scans within an average of 1.6 years were included. Regional annualized rates of flortaucipir uptake standardized uptake value ratios (SUVr) were calculated. Regional annualized rates of cortical volume change were measured with the Tensor Based Morphometry–Syn algorithm. Results The annual increase of flortaucipir SUVr was greater in the middle and superior occipital, fusiform, and inferior parietal cortices in DLB (mean: 0.017, 0.019, 0.019, and 0.015, respectively) compared with the CU (mean: −0.006, −0.009, −0.003, and − 0.005, respectively; P < 0.05). In patients with DLB (but not the CU), a longitudinal increase in flortaucipir SUVr was associated with longitudinal cortical atrophy rates in the lateral occipital and inferior temporoparietal cortices, hippocampus, and the temporal pole as well as a concurrent decline on Mini‐Mental State Examination and Clinical Dementia Rating–Sum of Boxes in the lateral occipital and the fusiform cortices. Conclusions Tau accumulation was faster in DLB compared with the CU, with increased accumulation rates in the lateral occipital and temporoparietal cortices. These increased rates of tau accumulation were associated with neurodegeneration and faster disease progression in DLB. Tau may be a potential treatment target in a subset of patients with DLB. © 2022 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society