PURPOSE:To investigate tumor mutation variations across different racial/ethnic groups to better understand implications for targeted cancer therapies. METHODS:A retrospective analysis of 5,045 patients at University of New Mexico Comprehensive Cancer Center who underwent tumor genetic testing between January 2015 and April 2022 was conducted. Data were standardized from internal genetic tests, FoundationOne, and Guardant next-generation sequencing panels. Chi-square tests, one-way analysis of variance, and negative binomial regression estimated differences in mutation rates across race/ethnicity, adjusting for cancer type, age, testing year, and number of genes screened. Primary outcomes included tumor mutation rates and their variation across racial/ethnic groups. Specific focus was placed on mutation frequencies in common genes, and association between race/ethnicity and mutations detected, adjusted for covariates. RESULTS:Among 5,045 patients-Hispanic/Latino (30%), American Indian (5.7%), Asian/Hawaiian Native (1.9%), Black (1.5%), non-Hispanic White (41%), and other/unknown (19.7%)-mutations were identified most commonly for Asian/Hawaiian Native individuals, with a rate of 0.068 mutations per gene screened (95% CI, 0.051 to 0.090), followed by White individuals (rate = 0.061, 95% CI, 0.051 to 0.072). Fewest mutations were identified for Black individuals, with a rate of 0.045 mutations per gene screened (95% CI, 0.033 to 0.061). Single-gene comparisons suggested BRAF mutations to be most prevalent in non-Hispanic Whites (5.8%, P = .015) while EGFR mutations were most common in Asian/Hawaiian Native patients (10.53%, P = .005). CONCLUSION:This study highlights substantial heterogeneity in tumor mutations across racial/ethnic groups while emphasizing the need for wider understanding of genomics and tailored approaches in cancer treatment. Findings underscore the need for equitable genomic testing, tailored therapies, and inclusive cancer care. Further research is necessary to bridge existing disparities, ensuring comprehensive, personalized cancer treatment for all patients.

The somatic JAK2V617F mutation (JAK2) in myeloproliferative neoplasms (MPNs) has been associated with an increased risk of thrombosis. Data are limited on the relationship between JAK2 and unexplained thrombophilia. As such, thrombophilia screening rarely includes JAK2 as a screening test, usually reserved for patients with thrombotic events in specific or unusual locations (e.g., mesenteric venous thrombosis). This study aims to determine the prevalence of JAK2 in patients with unexplained thrombophilia and its potential association in patients with cytosis. Patients who had previously undergone thrombophilia profile including molecular testing with DNA-stored at TriCore Reference Lab during the 2022 calendar year were identified (n=359). 124 patients were sequentially selected to undergo JAK2 testing. All 359 patients were screened to determine the presence of = 1 cytosis: white blood cell, red blood cell, or platelet count above the respective reference range’s upper limit of normal either within one month of thrombophilia screening OR evidence of persistent cytosis. 23 patients had cytosis (11 in the sequential cohort, 11 additional patients with DNA, 1 patient with insufficient DNA). 11 additional patients with cytosis were tested for JAK2. Quantitative RT-PCR results were assigned qualitative values using JAK2 DNA standard clinically validated thresholds: 0%-0.5% negative, 0.5%-1% indeterminate, and > 1% positive. Of the 135 samples tested for the JAK2, mean age was 46 (range of 1-86) years, and 103 (76.3%) were female. 1/135 (0.7%) samples was positive for JAK2. The single positive sample (32-year-old male) was within original sequential cohort without cytosis. A Fisher Exact Test shows that there is no correlation with cytosis and JAK2 status (p=1). These findings discourage using JAK2 in a first-line thrombophilia genetic screening panel, even in patients with concurrent cytosis. Future directions involve testing additional specimens to increase the sample size of random and cytosis groups. Clinical correlation would also potentially identify a additional associations with which patients could be screened for this testing.

Background: HLA homozygosity of specific alleles at a single locus is associated with increased risk for autoimmunity and/or more severe clinical phenotypes. However, the contribution of the overall limitation of HLA diversity across multiple loci to autoimmunity risk remains to be determined. Methods: We conducted a proof-of-concept case-control study of 413 individuals (279 cases with pediatric-onset autoimmune rheumatic diseases, 134 matched controls) examining the "Limitation of HLA Diversity" (LoHLAD) across multiple loci as an allele-independent risk factor for autoimmunity. We examined the association of LoHLAD with pediatric-onset autoimmune rheumatic diseases at five HLA loci (A, B, DQB1, DRB1, DRB3/4/5). LoHLAD was defined as (1) homozygosity at any of the examined loci, and/or (2) the presence of a single allele or the complete lack of an allele at the HLA-DRB3/4/5 locus. Results: The frequency of LoHLAD at any locus was significantly higher in cases compared to controls (65.95% vs. 30.60%, OR 4.39 [2.82-6.84], p < 0.0001). Higher frequencies of LoHLAD in cases compared to controls were observed at both class I (19.35% vs. 10.45%, OR 2.06 [1.10-3.86], p = 0.031) and class II (54.48% vs. 20.15%, OR 4.74 [2.92-7.69], p < 0.0001) loci. Specifically, significant differences between cases and controls were observed at the B (OR 8.63 [1.14-65.55], p = 0.016), DQB1 (OR 3.34 [1.27-8.78], p = 0.016), and DRB3/4/5 (OR 4.64 [2.77-7.75], p < 0.0001) loci. Multiple logistic regression models confirmed the ability of LoHLAD to positively predict autoimmunity. Conclusions: LoHLAD is a significant allele-independent risk factor for pediatric-onset autoimmune rheumatic disease.

HLA homozygosity of specific alleles at a single locus is associated with increased risk for autoimmunity. However, the contribution of the overall limitation of HLA allele diversity to autoimmunity risk remains to be determined. We conducted a proof-of-concept case-control study of 413 subjects (279 cases, 134 matched controls) examining the “Limitation of HLA Diversity” (LoHLAD) across multiple loci as an allele-independent risk factor for pediatric-onset autoimmune rheumatic disease. The association of LoHLAD with pediatric-onset autoimmune rheumatic diseases was examined at 5 HLA loci (HLA-A, HLA-B, HLA-DQB1, HLA- DRB1, HLA- DRB3/4/5). For the purpose of this study, we introduced a novel metric of LoHLAD defined as 1) homozygosity at any of the examined loci, and/or 2) the presence of a single allele or the complete lack of an allele at the HLA-DRB3/4/5 locus. The frequency of LoHLAD at any locus was significantly higher in cases compared to controls (65.95% vs 30.60%, OR 4.39 [2.82-6.84], P

Patient safety education is a mandated Common Program Requirement of the Accreditation Council for Graduate Medical Education and for the Royal College of Physicians and Surgeons of Canada in all medical residency and fellowship programs. Although many hospitals and healthcare environments have general patient safety education tools for trainees, few to none focus on the unique training milieu of pathologists, including a mix of highly automated and manual error-prone processes, frequent multiplicity of events, and lack of direct patient relationships for error disclosure. We established a national Association of Pathology Chairs -Program Directors Section Workgroup focused on patient safety education for pathology trainees entitled Training Residents in Patient Safety (TRIPS). TRIPS included diverse representatives from across the United States, as well as representatives from pathology organizations including the American Board of Pathology, the American Society for Clinical Pathology, the United States and Canadian Academy of Pathology, the College of American Pathologists, and the Society to Improve Diagnosis in Medicine. Objectives of the workgroup included developing a standardized patient safety curriculum, designing teaching and assessment tools, and refining them with pilot sites. Here we report the establishment of TRIPS as well as data from national needs assessment of Program Directors across the country, who confirmed the need for a standardized patient safety curriculum.

Primary myelofibrosis (PMF) is a clonal myeloproliferative neoplasm driven by canonical gene mutations in JAK2, CALR, or MPL in >80% of the cases. PMF that lacks these canonical alterations is termed triple-negative PMF (TN-PMF). The pathologic and genetic characteristics of TN-PMF compared with those of conventional PMF with canonical driver mutations (DM-PMF) have not been well studied. We aimed to identify clinicopathologic and molecular genetic differences between patients with TN-PMF (n = 56) and DM-PMF (n = 89), all of whom fulfilled the 2016 World Health Organization diagnostic criteria for PMF. Compared with the control group, patients in the TN-PMF group were more likely to have thrombocytopenia and less likely to have organomegaly. The bone marrow in patients with TN-PMF showed fewer granulocytic elements and more frequent dyserythropoiesis. Cytogenetic analysis showed a higher incidence of trisomy 8. Targeted next-generation sequencing revealed a lower frequency of ASXL1 mutations but enrichment of ASXL1/SRSF2 comutations. Our findings demonstrated several clinicopathologic and molecular differences between TN-PMF and DM-PMF. These findings, particularly the observed mutation profile characterized by a higher frequency of ASXL1 and SRSF2 comutation, suggest that at least a subset of TN-PMF may be pathogenetically different from DM-PMF, with potential prognostic implications.

In this review of megaloblastic anemia (MA), an overview of vitamin B12 and folate body requirements, biochemical pathways, and laboratory testing strategies will be provided. However, the focus of this review is the classic and unique features of MA in blood and bone marrow. Acquired MA is a benign disorder for many, but can be detrimental for some. The clinical presentation can vary considerably, and the spectrum of symptoms and signs is diverse and quite broad. Prompt recognition and therapy are critical to prevent potential irreversible damage and clinical sequelae, especially in patients with vitamin B12 deficiency. A delay in diagnosis of vitamin B12 deficiency can result in significant neurologic sequelae that may not fully resolve with treatment, including in neonates and young infants. The blood and bone marrow features in MA can closely mimic thrombocytopenic purpura, myelodysplasia, and other myeloid neoplasms. Both pancytopenia and normal MCV at presentation are common in MA and raise unique challenges for the diagnostician. Partially treated MA is also a significant diagnostic "trap". MA is highly responsive to treatment, and patients tend to improve rapidly upon treatment initiation. However, the broad range of clinical and hematologic features makes the rapid, successful diagnosis of MA a unique challenge for the hematopathologist. Even in the era of state-of-the-art laboratory testing, a high suspicion is required.

Developments in genomics are profoundly influencing medical practice. With increasing use of genetic and genomic testing across every aspect of the health care continuum, patients and their families are increasingly turning to primary care physicians (PCPs) for discussion and advice regarding tests, implications, and results. Yet, with the rapid growth of information, technology, and applications, PCPs are finding it challenging to fill the gaps in knowledge and support the growing needs of their patients. A critical component in expanding PCP genomic literacy lies in the education of physicians in training and in practice. Although a framework for developing physician competencies in genomics has already been developed, the Association for Molecular Pathology is uniquely situated to actively utilize the skills of its members to engage and support PCPs in this effort. This report provides an overview and a suggested basic teaching framework, which can be used by molecular professionals in their individual institutions as a starting point for educational outreach.

Pathogenic New World orthohantaviruses cause hantavirus cardiopulmonary syndrome (HCPS), a severe immunopathogenic disease in humans manifested by pulmonary edema and respiratory distress, with case fatality rates approaching 40%. High levels of inflammatory mediators are present in the lungs and systemic circulation of HCPS patients. Previous studies have provided insights into the pathophysiology of HCPS. However, the longitudinal correlations of innate and adaptive immune responses and disease outcomes remain unresolved. This study analyzed serial immune responses in 13 HCPS cases due to Sin Nombre orthohantavirus (SNV), with 11 severe cases requiring extracorporeal membrane oxygenation (ECMO) treatment and two mild cases. We measured viral load, levels of various cytokines, urokinase plasminogen activator (uPA), and plasminogen activator inhibitor-1 (PAI-1). We found significantly elevated levels of proinflammatory cytokines and PAI-1 in five end-stage cases. There was no difference between the expression of active uPA in survivors’ and decedents’ cases. However, total uPA in decedents’ cases was significantly higher compared to survivors’. In some end-stage cases, uPA was refractory to PAI-1 inhibition as measured by zymography, where uPA and PAI-1 were strongly correlated to lymphocyte counts and IFN-γ. We also found bacterial co-infection influencing the etiology and outcome of immune response in two cases. Unsupervised Principal Component Analysis and hierarchical cluster analyses resolved separate waves of correlated immune mediators expressed in one case patient due to a sequential co-infection of bacteria and SNV. Overall, a robust proinflammatory immune response, characterized by an imbalance in T helper 17 (Th17) and regulatory T-cells (Treg) subsets, was correlated with dysregulated inflammation and mortality. Our sample size is small; however, the core differences correlated to survivors and end-stage HCPS are instructive.

Liquid based cytology (LBC) specimens are increasingly utilized for molecular analysis, as results are comparable to molecular analysis performed on traditional specimens (biopsy or cell block). However, there are few studies demonstrating the long‐term viability of DNA in LBC samples.

You have accessJournal of UrologyTransplantation & Vascular Surgery: Renal Transplantation & Vascular Surgery II (MP70)1 Apr 2019MP70-16 PRELIMINARY IDENTIFICATION OF IMMUNE CELLS IN THE PERFUSATE OF RENAL ALLOGRAFTS UNDERGOING HYPOTHERMIC MACHINE PERFUSION Jordan Foreman*, Devon Chabot-Richards, Barbara Masten, Julie Riley, and Michael Davis Jordan Foreman*Jordan Foreman* More articles by this author , Devon Chabot-RichardsDevon Chabot-Richards More articles by this author , Barbara MastenBarbara Masten More articles by this author , Julie RileyJulie Riley More articles by this author , and Michael DavisMichael Davis More articles by this author View All Author Informationhttps://doi.org/10.1097/01.JU.0000557108.54232.e9AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVES: Hypothermic machine perfusion (HMP) in renal allografts have shown improved outcomes and reduced rejection in recipients. We hypothesize that HMP washes out antigen presenting cells (APC), our study is designed to determine the concentrations of immune cells in perfusate fluid. METHODS: Renal allografts on HMP transplanted at our institution between 5/2016 to 2/2017 were included in our study. After the allograft was removed from the pump, the perfusate fluid was collected sterilely and analyzed using flow cytometry. Concentrations of immune cells including B cells, T cells, monocytes, granulocytes, NK cells, and dendritic cells were examined. RESULTS: Twenty-one samples were obtained (11 right kidneys, 10 left kidneys). The most common cell type was CD8 T cells (57.8%, range 19-84.7). The remaining cell types, mean concentration and range were CD15 granulocytes (48.6%, 15.1-82.8), CD4 T cells (42.3, 15.3-81), CD14 monocytes (15.8, 5.3-31.6), CD56 NK cells (8.2, 3.3-16.6), CD11c dendritic cells (6.3, 0-20.8), CD 123 dendritic cells (3.2, 0-19.4), and CD 20 B cells (3.0, 0.1-17). There was no statistical difference in sidedness (p=0.9222) or between donation after cardiac/anoxia and brain death donors (p=0.9220). CONCLUSIONS: The most common immune cell type in perfusate fluid was CD8 T cells followed by CD15 granulocytes and CD14 monocytes. Granulocytes and monocytes are common APC and stimulate T cells to become CD8 T cells, these cells are certainly involved in antibody-mediated rejection in the recipient. These concentrations are higher than expected levels and suggest that a “wash out” is occurring. Future studies are aimed at determining cellular concentration in cold storage as well as ways to increase filtration of APC in donors prior to renal transplantation and the effect on recipient outcomes. Source of Funding: UNMH Surgery Research Grant Albuquerque, NM© 2019 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 201Issue Supplement 4April 2019Page: e1044-e1044 Advertisement Copyright & Permissions© 2019 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jordan Foreman* More articles by this author Devon Chabot-Richards More articles by this author Barbara Masten More articles by this author Julie Riley More articles by this author Michael Davis More articles by this author Expand All Advertisement PDF downloadLoading ...

Introduction: In the current era of precision medicine, CDx tests have become the basis for optimal patient care and targeted treatment. While technical aspects of a CDx assay's performance are audited regularly by participation in external quality assessment (EQA) programs, the reporting of these CDx markers is rarely scrutinized. The aim of this first of its kind CDx report program is to get a detailed picture of hemato-oncology CDx marker reporting across multiple countries. Therefore, we evaluated multiple laboratory parameters and reviewed actual content of clinical CDx reports. Methods: Thirty-four clinical labs from Germany, Italy, Spain, and the United Kingdom provided information for 7 biomarkers used for treatment decisions in hemato-oncology. An identical program is already running in Australia and the United States and will be initiated shortly in India and China. The CDx markers covered in this CDx report program were: BCR-ABL1 in chronic myeloid leukemia (CML), at diagnosis; BCR-ABL1 in CML, minimal residual disease (MRD); IDH1/2 in acute myeloid leukemia (AML); FLT3-ITD in AML; FLT3-TKD in AML; IGHV in chronic lymphocytic leukemia (CLL); and TP53 in CLL. The information requested from participating laboratories included two anonymized or blank reports: one with a positive/mutant result and one with a negative/wild-type result. The received anonymized reports were reviewed by experts within each country according to pre-agreed criteria. In addition, labs participated in a short online survey evaluating test volumes, turnaround times (TATs), positivity rates, participation in an EQA program, and status regarding accreditation and reimbursement. The results of the questionnaires were forwarded as anonymized and aggregated data to reviewing experts. Results: Overall, we received 184 survey datasets and 179 sets of anonymized reports. The review of the anonymized reports according to pre-defined criteria revealed differences in the way CDx results are represented and interpreted in clinical reports. Since not all markers covered in this program were tested by all participating labs, the number of survey datasets per CDx marker ranged from 16 to 34 (IGHV: 16; TP53: 19; IDH1/2: 24; FLT3-TKD: 28; FLT3-ITD: 30; BCR-ABL1 [MRD]: 33; and BCR-ABL1 [diagnosis]: 34). The 184 survey datasets represented more than 7000 tests per month (Figure). The stated average TAT across all covered markers was 6.9 days, ranging from 5.3 days to 8.6 days in the covered countries. The TATs for the individual markers ranged from 4.4 days to 9.4 days (FLT3-ITD: 4.4 days; FLT3-TKD: 4.6 days; BCR-ABL1 [diagnosis]: 5.3 days; BCR-ABL1 [MRD]: 6.9 days; IDH1/2: 8.4 days; IGHV: 9.3 days; and TP53: 9.4 days). In 103/179 (58%) datasets labs participated in EQA programs, and in 76/179 (42%) datasets labs did not participate in EQA programs. For 84/180 (47%) datasets, labs were ISO15189-accredited; for 12/180 (7%) datasets, labs were College of American Pathologists (CAP)-accredited; and for 84/180 (47%) datasets, labs were not accredited by either ISO15189 or CAP. Conclusion: CDx report program results reveal a broad range globally in CDx reporting practices. The identified differences in laboratory parameters, such as TAT and the actual content of clinical CDx reports, suggest a need for international harmonization of CDx reporting. The anonymized and aggregated data generated provide the basis for other initiatives and may support guideline updates and the international harmonization of CDx reporting. Figure. Estimated number of companion diagnostic tests per month covered in the CDx report program. Figure Disclosures Delic: Diaceutics: Employment, Equity Ownership. Blombery:Janssen: Honoraria; Novartis: Consultancy; Invivoscribe: Honoraria. Calasanz:Janssen: Honoraria; Diaceutics: Honoraria; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Honoraria. Colomer:Novartis: Honoraria; Incyte: Honoraria. Evans:Diaceutics: Honoraria; Novartis: Honoraria. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Mason:Novartis: Honoraria; Jazz Pharmaceuticals: Honoraria; AbbVie: Honoraria. Thiede:Daiichi Sankyo: Honoraria; Diaceutics: Membership on an entity's Board of Directors or advisory committees; Novartis: Consultancy, Honoraria, Research Funding, Speakers Bureau; AgenDix GmbH: Employment, Equity Ownership. Clark:Diaceutics: Employment, Equity Ownership.

The human leukocyte antigen (HLA) system is a highly polymorphic family of genes involved in immunity and responsible for identifying self versus non-self. HLA typing is essential for solid organ and bone marrow transplantation as well as in non-transplant settings such as disease association and pharmacogenomics. Typing of HLA genes differs from most molecular testing as, rather than evaluating differences from an accepted “wild-type” gene, it must distinguish between thousands of similar, but distinct alleles. This article will describe the HLA system and nomenclature. We will then discuss clinical uses of HLA typing including solid organ transplantation, hematopoietic stem cell transplantation, evaluation of platelet refractory patients, disease association, and pharmacogenetics. Finally, we describe common molecular methods of HLA typing.

Renal allografts stored with hypothermic machine perfusion (HPM) have improved outcomes compared to static methods. Passenger immune cells from the allograft have been implicated in acute rejection; improved outcomes with HPM may be due to cell removal by the pump. This study aimed to identify passenger immune cells present in the perfusate of HPM-preserved allografts. Once characterized, future strategies to suppress these cells may prevent acute rejection and improve graft survival. All kidneys placed on HPM prior to transplant from May 2016 to August 2017 were eligible. Perfusate fluid was analyzed using flow cytometry for immune cells and information about the donors was collected. Wilcoxon Scores and Spearman Correlation Coefficient were used in statistical analysis. Twenty-seven perfusate specimens were analyzed. Selected donor characteristics: 63.6% male, 36.4% female; 63.6% donation after cardiac death; 54.5% pre-procurement transfusion. The mean HPM and cold ischemia times (CIT) were 9.9 h and 13.9 h respectively, and mean KDPI 36%. Flow cytometry showed a heterogeneous mix of immune cell types in the perfusate, similar to blood. CD15+ granulocytes, CD14+ monocytes, and CD8+ T cells were predominant in the perfusate. Dendritic and natural killer (NK) cells in perfusate were more abundant compared to blood. Passenger immune cells, specifically dendritic cells have been implicated in acute rejection. This study showed the perfusate of renal allografts contains an array of immune cells, with a significantly higher percentage of dendritic cells and NK cells, as compared to blood. Future studies would show if HPM removal of these immune cells accounts for improved outcomes. ∗Frequencies of Cell Types in Human Peripheral Blood, www.stemcell.com.