The American Society for Apheresis (ASFA) Journal of Clinical Apheresis (JCA) Special Issue Writing Committee is charged with reviewing, updating, and categorizing indications for the evidence-based use of therapeutic apheresis in human disease. Since the 2007 JCA Special Issue (Fourth Edition), the Committee has incorporated systematic review and evidence-based approaches in the grading and categorization of apheresis indications. This Seventh Edition of the JCA Special Issue continues to maintain this methodology and rigor to make recommendations on the use of apheresis in a wide variety of diseases/conditions. The JCA Seventh Edition, like its predecessor, has consistently applied the category and grading system definitions in the fact sheets. The general layout and concept of a fact sheet that was used since the fourth edition has largely been maintained in this edition. Each fact sheet succinctly summarizes the evidence for the use of therapeutic apheresis in a specific disease entity. The Seventh Edition discusses 87 fact sheets (14 new fact sheets since the Sixth Edition) for therapeutic apheresis diseases and medical conditions, with 179 indications, which are separately graded and categorized within the listed fact sheets. Several diseases that are Category IV which have been described in detail in previous editions and do not have significant new evidence since the last publication are summarized in a separate table. The Seventh Edition of the JCA Special Issue serves as a key resource that guides the utilization of therapeutic apheresis in the treatment of human disease. J. Clin. Apheresis 31:149-162, 2016. © 2016 Wiley Periodicals, Inc.

Guidelines on the Use of Therapeutic Apheresis in Clinical Practice—Evidence-Based Approach from the Writing Committee of the American Society for Apheresis: The Sixth Special Issue

Several genetic alterations characteristic of leukemia and lymphoma have been detected in the blood of individuals without apparent hematological malignancies. The Cancer Genome Atlas (TCGA) provides a unique resource for comprehensive discovery of mutations and genes in blood that may contribute to the clonal expansion of hematopoietic stem/progenitor cells. Here, we analyzed blood-derived sequence data from 2,728 individuals from TCGA and discovered 77 blood-specific mutations in cancer-associated genes, the majority being associated with advanced age. Remarkably, 83% of these mutations were from 19 leukemia and/or lymphoma-associated genes, and nine were recurrently mutated (DNMT3A, TET2, JAK2, ASXL1, TP53, GNAS, PPM1D, BCORL1 and SF3B1). We identified 14 additional mutations in a very small fraction of blood cells, possibly representing the earliest stages of clonal expansion in hematopoietic stem cells. Comparison of these findings to mutations in hematological malignancies identified several recurrently mutated genes that may be disease initiators. Our analyses show that the blood cells of more than 2% of individuals (5-6% of people older than 70 years) contain mutations that may represent premalignant events that cause clonal hematopoietic expansion.

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Age-related mutations associated with clonal hematopoietic expansion and malignancies

Single-cell RNA-seq enables the quantitative characterization of cell types based on global transcriptome profiles. We present single-cell consensus clustering (SC3), a user-friendly tool for unsupervised clustering, which achieves high accuracy and robustness by combining multiple clustering solutions through a consensus approach (http://bioconductor.org/packages/SC3). We demonstrate that SC3 is capable of identifying subclones from the transcriptomes of neoplastic cells collected from patients.

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SC3: consensus clustering of single-cell RNA-seq data

Recent genomic studies have identified novel recurrent somatic mutations in patients with myeloid malignancies, including myeloproliferative neoplasms (MPNs), myelodysplastic syndrome (MDS) and acute myeloid leukaemia (AML). In some cases these mutations occur in genes with known roles in regulating chromatin and/or methylation states in haematopoietic progenitors, and in other cases genetic and functional studies have elucidated a role for specific mutations in altering epigenetic patterning in myeloid malignancies. In this Review we discuss recent genetic and functional data implicating mutations in epigenetic modifiers, including tet methylcytosine dioxygenase 2 (TET2), isocitrate dehydrogenase 1 (IDH1), IDH2, additional sex combs-like 1 (ASXL1), enhancer of zeste homologue 2 (EZH2) and DNA methyltransferase 3A (DNMT3A), in the pathogenesis of MPN, MDS and AML, and discuss how this knowledge is leading to novel clinical, biological and therapeutic insights.

The role of mutations in epigenetic regulators in myeloid malignancies

The American Society for Apheresis (ASFA) JCA Special Issue Writing Committee is charged with reviewing, updating and categorizating indications for therapeutic apheresis. Beginning with the 2007 ASFA Special Issue (Fourth Edition), the committee has incorporated systematic review and evidence‐based approach in the grading and categorization of indications. This Sixth Edition of the ASFA Special Issue has further improved the process of using evidence‐based medicine in the recommendations by consistently applying the category and GRADE system definitions, but eliminating the “level of evidence” criteria (from the University HealthCare Consortium) utilized in prior editions given redundancy between GRADE and University HealthCare Consortium systems. The general layout and concept of a fact sheet that was utilized in the Fourth and Fifth Editions, has been largely maintained in this edition. Each fact sheet succinctly summarizes the evidence for the use of therapeutic apheresis in a specific disease entity. This article consists of 78 fact sheets (increased from 2010) for therapeutic indications in ASFA categories I through IV, with many diseases categorized having multiple clinical presentations/situations which are individually graded and categorized. J. Clin. Apheresis 28:145–284, 2013. © 2013 Wiley Periodicals, Inc.

Guidelines on the use of therapeutic apheresis in clinical practice - Evidence-based approach from the writing committee of the american society for apheresis

Background Somatic mutations in the Janus kinase 2 gene (JAK2) occur in many myeloproliferative neoplasms, but the molecular pathogenesis of myeloproliferative neoplasms with nonmutated JAK2 is obscure, and the diagnosis of these neoplasms remains a challenge. Methods We performed exome sequencing of samples obtained from 151 patients with myeloproliferative neoplasms. The mutation status of the gene encoding calreticulin (CALR) was assessed in an additional 1345 hematologic cancers, 1517 other cancers, and 550 controls. We established phylogenetic trees using hematopoietic colonies. We assessed calreticulin subcellular localization using immunofluorescence and flow cytometry. Results Exome sequencing identified 1498 mutations in 151 patients, with medians of 6.5, 6.5, and 13.0 mutations per patient in samples of polycythemia vera, essential thrombocythemia, and myelofibrosis, respectively. Somatic CALR mutations were found in 70 to 84% of samples of myeloproliferative neoplasms with nonmutated JAK2, in 8...

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Somatic CALR Mutations in Myeloproliferative Neoplasms with Nonmutated JAK2

<h3>Introduction</h3> Radiofrequency ablation (RFA) is the treatment of choice for dysplastic Barrett’s oesophagus (BO), however recurrence occurs in up to 25% of cases, usually at the gastro-oesophageal junction (GOJ). We have shown that a multi-gene methylation panel (ZNF345, TFPI2 and ZNF569) can differentiate BO from normal GOJ epithelium. This study aims to determine whether this methylation panel can predict risk of relapse post-RFA. <h3>Methods</h3> This is a prospective cohort study on patients who achieved endoscopic remission post-RFA. Patients were eligible if they had 1) no endoscopic evidence of BO or islands ≥5mm, 2) no suspicious dysplastic lesion at GOJ, and 3) no histological evidence of oesophageal intestinal metaplasia (IM) at first post-RFA follow-up (GOJ IM allowed). Patients received endoscopy at baseline, 6 and 12 months. Methylation score (Meth-score) was assessed by Methylight assay on random or targeted GOJ biopsies. The primary outcome was the correlation of Meth-score and histological outcome. The secondary outcome was the correlation of total baseline methylation level and dysplastic recurrence at follow-up. <h3>Results</h3> We included 56 patients with a total of 90 endoscopies at various timepoints of follow-up. Mean age, circumferential and maximum length of BO pre-RFA was 68.6 years (standard deviation, SD 7.5), 2.2cm (SD 3.0) and 4.4cm (SD 3.0), respectively. Seven (13%), 28 (50%) and 21 (38%) patients had pre-RFA low-grade dysplasia (LGD), high-grade dysplasia (HGD) and intramucosal carcinoma (IMC), respectively. Meth-score comparing patients with gastric metaplasia (GOJ-GM, n=60) and intestinal metaplasia (GOJ-IM, n=25) was 1.2% vs 36.9%, p&lt;0.001, and between GOJ-GM and dysplasia (n=5) was 1.2% vs 43.4%, p&lt;0.001. When assessing baseline Meth-score stratified by highest histology at any time during follow-up, patients with non-dysplastic recurrence (n=50) had a lower total baseline methylation compared to dysplastic recurrence (n=6), with Meth-score of 8.5% vs 39.0%, p=0.03. <h3>Conclusion</h3> A multi-gene methylation panel can discriminate patients with GM, IM and dysplasia on GOJ biopsies. Patients with stable histological remission (no IM or dysplasia at follow-up) had low baseline methylation levels. This biomarker panel can risk-stratify patients so that stricter surveillance is offered only to those with higher total methylation levels who are more likely to develop recurrence. 

M. Bianchi

Papers

Somatic CALR Mutations in Myeloproliferative Neoplasms with Nonmutated JAK2

P140 The utility of a multi-gene methylation panel to risk-stratify patients with Barrett’s oesophagus after radiofrequency ablation