Immunophenotypic assessment of clonal plasma cells and B-cells in bone marrow and blood in the diagnostic classification of early stage monoclonal gammopathies : an iSTOPMM study

Publisher Copyright: © 2023, The Author(s).Monoclonal gammopathy of undetermined significance (MGUS) is the earliest discernible stage of multiple myeloma (MM) and Waldenström’s macroglobulinemia (WM). Early diagnosis of MG may be compromised by the low-level infiltration, undetectable to low-sensitive methodologies. Here, we investigated the prevalence and immunophenotypic profile of clonal (c) plasma cells (PC) and/or cB-lymphocytes in bone marrow (BM) and blood of subjects with a serum M-component from the iSTOPMM program, using high-sensitive next-generation flow cytometry (NGF), and its utility in the diagnostic classification of early-stage MG. We studied 164 paired BM and blood samples from 82 subjects, focusing the analysis on: 55 MGUS, 12 smoldering MM (SMM) and 8 smoldering WM (SWM). cPC were detected in 84% of the BM samples and cB-lymphocytes in 45%, coexisting in 39% of cases. In 29% of patients, the phenotypic features of cPC and/or cB-lymphocytes allowed a more accurate disease classification, including: 19/55 (35%) MGUS, 1/12 (8%) SMM and 2/8 (25%) SWM. Blood samples were informative in 49% of the BM-positive cases. We demonstrated the utility of NGF for a more accurate diagnostic classification of early-stage MG.Peer reviewe

Determining hemodilution in diagnostic bone marrow aspirated samples in plasma cell disorders by next-generation flow cytometry : Proposal for a bone marrow quality index

Publisher Copyright: © 2023, The Author(s).Hemodilution of bone marrow (BM) aspirates is a limitation of multiparameter flow cytometry (MFC) in plasma cell disorders. There is a need for a validated approach for assessing sample quality and the distribution of non-plasma cell BM populations by MFC could provide a solution. We evaluated BM-associated cell populations, assessed by next-generation flow cytometry (NGF) and white blood cell (WBC) count in 351 BM aspirated samples from 219 participants with plasma cell disorders in the Iceland Screens, Treats, or Prevents MM study (iStopMM), as markers of hemodilution by their discriminatory ability between first and (generally more hemodiluted) second pull BM aspirated samples. The most discriminating markers were used to derive a novel BM quality index (BMQI). Nucleated red blood cells and myeloid precursors provided the greatest discriminatory ability between first vs second pull samples (area under the curve (AUC): 0.87 and 0.85, respectively), significantly better than B cell precursors (AUC = 0.64; p < 0.001), mast cells (AUC = 0.65; p < 0.001), and the BM WBC count (AUC = 0.77; p < 0.05). We generated a novel BMQI that is intrinsic to current NGF protocols, for evaluating quality of diagnostic BM samples and suggest the use of a BMQI scoring system for interpreting results and guiding appropriate actions.Peer reviewe

Iceland screens, treats, or prevents multiple myeloma (iStopMM): a population-based screening study for monoclonal gammopathy of undetermined significance and randomized controlled trial of follow-up strategies.

To access publisher's full text version of this article, please click on the hyperlink in Additional Links field or click on the hyperlink at the top of the page marked DownloadMonoclonal gammopathy of undetermined significance (MGUS) precedes multiple myeloma (MM). Population-based screening for MGUS could identify candidates for early treatment in MM. Here we describe the Iceland Screens, Treats, or Prevents Multiple Myeloma study (iStopMM), the first population-based screening study for MGUS including a randomized trial of follow-up strategies. Icelandic residents born before 1976 were offered participation. Blood samples are collected alongside blood sampling in the Icelandic healthcare system. Participants with MGUS are randomized to three study arms. Arm 1 is not contacted, arm 2 follows current guidelines, and arm 3 follows a more intensive strategy. Participants who progress are offered early treatment. Samples are collected longitudinally from arms 2 and 3 for the study biobank. All participants repeatedly answer questionnaires on various exposures and outcomes including quality of life and psychiatric health. National registries on health are cross-linked to all participants. Of the 148,704 individuals in the target population, 80 759 (54.3%) provided informed consent for participation. With a very high participation rate, the data from the iStopMM study will answer important questions on MGUS, including potentials harms and benefits of screening. The study can lead to a paradigm shift in MM therapy towards screening and early therapy.Black Swan Research Initiative by the International Myeloma Foundation Icelandic Centre for Research European Research Council (ERC) University of Iceland Landspitali University Hospita

Áhrif efnaþátta og efnasambanda úr sjávarhryggleysingjum á ónæmissvör

Natural products are a productive and well recognized source of new therapeutics and much recent interest has been on exploring the vast biodiversity of marine organisms in search of novel bioactive compounds. The pathobiology of an array of autoimmune and inflammatory disorders is characterized by dysregulation of immune responses, leading to a chronic state of inflammation. Modulation of immune responses by bioactive natural compounds could, therefore, be of therapeutic value in the treatment of pathological inflammation. Antigen presenting cells (APCs), in particular dendritic cells (DCs), are capable of polarizing T cells towards specific effector phenotypes dependent on the nature of their activation and are a prime target for evaluating immunomodulating effects of natural compounds in the search for effective immunotherapeutic agents. The aim of this study was to identify and isolate anti-inflammatory compounds from marine invertebrates using a bioassay-guided isolation procedure and characterize the immunomodulatory effects of obtained bioactive fractions/compounds on DCs and DC mediated T cell activation. Human monocyte-derived DCs were matured and activated by lipopolysaccharide (LPS), tumour necrosis factor (TNF)-α and interleukin (IL)-1β and cultured in the presence of fractions/compounds from the sponge (Porifera) Halichondria sitiens (H. sitiens) for 24 h. The effects of fractions/compounds on DC maturation was evaluated by the expression of the antigen-presentation and co-stimulation molecules CD86 and HLA-DR, respectively, and the anti-inflammatory properties were evaluated by measuring concentrations of the pro-inflammatory cytokines IL-12p40 and IL-6 and the anti-inflammatory/regulatory cytokines IL-10 and IL-27. The effects of H. sitiens fractions on DC mediated T cell activation was evaluated by measuring secretion of T helper (TH) cell specific cytokines, interferon (IFN)-γ (TH1), IL-13 (TH2), IL-17 (TH17) and IL-10 (regulatory T cell, Treg), by allogeneic CD4+ T cells activated by DCs which had been treated with selected bioactive fractions. Fractionation of H. sitiens resulted in a number of bioactive fractions but not in the isolation of pure bioactive compounds. The bioactive fractions contained mostly lipid compounds and had different effects on cytokine secretion by DCs, reducing and/or enhancing secretion of pro- and anti-inflammatory cytokines, respectively. The H. sitiens fractions did not affect maturation of the DCs, as was evident by unaltered expression of the surface molecules, CD86 and HLA-DR. These molecules are important for effective activation of T cells and DCs matured in the presence of fractions from H. sitiens are, therefore, expected to have unimpaired T cell activation capacities. Most of the bioactive fractions inhibited secretion of the pro-inflammatory cytokines IL-12p40 and IL-6 and two fractions enhanced secretion of the anti-inflammatory cytokine IL-10. Furthermore, allogeneic CD4+ T cells activated by DCs treated with fraction B3b3-J, which reduced IL-12p40 and IL-6 production by DCs, secreted less of the TH1 specific cytokine IFN-γ and more of the TH2 specific cytokine IL-13, indicating a TH2 biased differentiation of the T cells. Two of the fractions from H. sitiens induced morphological changes in the DCs. These changes were characterized by extreme elongation of the DC dendrites. Taken together, these results show that H. sitiens contains bioactive compounds that are capable of affecting secretion of both pro- and anti-inflammatory cytokines by DCs and modulate DC mediated T cell activation. These compounds could, therefore, have immunotherapeutic potential in the treatment of inflammatory disorders.Náttúruefni eru mikilvæg uppspretta nýrra lyfja og síðustu áratugi hefur verið lögð aukin áhersla á nýtingu sjávarlífvera í leit að lífvirkum efnum til þróunar á lyfjasprotum. Langvarandi ræsing á ónæmissvörum getur leitt af sér króníska bólgu sem er einkennandi þáttur í meingerð fjölda sjálfsofnæmis- og bólgusjúkdóma og því geta lífvirk efni sem tempra ónæmissvör verið mögulegt meðferðarúrræði gegn langvarandi bólgu. Sýnifrumur, þá sérstaklega angafrumur, stjórna sérhæfingu óreyndra T frumna í mismunandi gerðir verkfrumna sem hafa hlutverk ýmist í miðlun eða temprun á ónæmissvörum. Þetta sérhæfingarferli er háð því umhverfi sem angafrumurnar ræsast í og leiðir af sér viðeigandi ónæmissvör undir eðlilegum kringumstæðum. Með því að skoða tjáningu á einkennissameindum fyrir ræstar angafrumur og seytingu á ónæmisstýrandi boðefnum er hægt að meta ónæmistemprandi áhrif náttúruefna í leit að bólguhamlandi lyfjasprotum. Markmið rannsóknarinnar var að greina og einangra bólguhamlandi efni úr sjávarhryggleysingjum með notkun lífvirknileiddrar einangrunar og skilgreina ónæmistemprandi áhrif lífvirkra efnaþátta/efnasambanda á angafrumur og angafrumu-miðlaða ræsingu á T frumum. Smáætur (monocytes) voru einangraðar úr blóði og sérhæfðar í angafrumur sem síðan voru þroskaðar og ræstar með lípópólýsakkaríði (LPS), tumour necrosis factor (TNF)-α og interleukin (IL)-1β samhliða því sem þær voru ræktaðar með efnaþáttum/efnasamböndum einangruðum úr Halichondria sitiens (H. sitiens) svampi (Porifera) í 24 klst. Áhrif efnaþátta/efnasambanda á þroskun angafrumna voru metin með því að mæla tjáningu á sýni- og hjálparboðssameindunum HLA-DR og CD86 á yfirborði angafrumna og ónæmistemprandi áhrif metin með mælingu á styrk bólguboðefnanna IL-12p40 og IL-6 og bólguhamlandi/ónæmisstýrandi boðefnanna IL-10 og IL-27 í frumufloti. Áhrif efnaþátta á angafrumu-miðlaða ræsingu á CD4+ T frumum voru metin með mælingu á styrk boðefna sem einkenna mismunandi T hjálparfrumusvipgerðir, interferon (IFN)-γ (TH1), IL-13 (TH2), IL-17 (TH17) og IL-10 (Treg), í floti úr samrækt angafrumna sem höfðu verið meðhöndlaðar með völdum lífvirkum þáttum og ósamgena T frumna. Þáttun á H. sitiens leiddi af sér fjölda lífvirkra efnaþátta en skilaði ekki einangrun á hreinu lífvirku efni. Lífvirku þættirnir innihéldu að mestu lípíð og höfðu þeir áhrif á seytingu angafrumna á boðefnum, ýmist til minnkunar og/eða aukningar á bólgu- og bólguhamlandi boðefnum. H. sitiens efnaþættirnir höfðu ekki áhrif á þroskun angafrumna þar sem tjáning á yfirborðssameindunum HLA-DR og CD86 hélst óbreytt. Tjáning á þessum sameindum er mikilvæg fyrir ræsingu T frumna og því ættu lífvirku H. sitiens efnaþættirnir ekki að hafa áhrif á ræsigetu angafrumna. Flestir lífvirku þættirnir drógu úr seytingu á bólguboðefnunum IL-12p40 og IL-6 og tveir þættir juku seytingu á bólguhamlandi boðefninu IL-10. Ósamgena CD4+ T frumur sem ræstar voru af angafrumum sem höfðu verið meðhöndlaðar með þætti B3b3-J, sem dró úr seytingu angafrumna á IL-12p40 og IL-6, seyttu minna af TH1 boðefninu IFN-γ og meira af TH2 boðefninu IL-13, sem bendir til að ónæmissvarið sveigist frá TH1 yfir í TH2 sérhæfingu á T frumum. Tveir þættir höfðu áhrif á formgerð angafrumna sem einkenndust af töluverðri ílengingu á öngum frumnanna. Samantekið sýna þessar niðurstöður að H. sitiens svampurinn inniheldur lífvirk efnasambönd sem hafa áhrif á bæði bólgumiðlandi og bólguhamlandi boðefnaseytingu angafrumna og á angafrumu-miðlaða ræsingu á T frumum. Þessi efnasambönd gætu því haft gildi við meðferð á langvarandi bólgu

Correction : Iceland screens, treats, or prevents multiple myeloma (iStopMM): a population-based screening study for monoclonal gammopathy of undetermined significance and randomized controlled trial of follow-up strategies (Blood Cancer Journal, (2021), 11, 5, (94), 10.1038/s41408-021-00480-w)

Publisher Copyright: © The Author(s) 2023.In this article the author name Malin Hultcrantz was incorrectly written as Malin Hulcrantz. The original article has been corrected

Iceland screens, treats, or prevents multiple myeloma (iStopMM) : a population-based screening study for monoclonal gammopathy of undetermined significance and randomized controlled trial of follow-up strategies

Funding Information: P.K. is an employee of The Binding Site. BGMD has done consultancy for Amgen, Janssen, Celgene, Takeda. S.H. is the director of The Binding Site. O.L. has received research funding from: National Institutes of Health (NIH), National Cancer Institute (NCI), U.S. Food and Drug Administration (FDA), Multiple Myeloma Research Foundation (MMRF), International Myeloma Foundation (IMF), Leukemia and Lymphoma Society (LLS), Perelman Family Foundation, Rising Tide Foundation, Amgen, Celgene, Janssen, Takeda, Glenmark, Seattle Genetics, Karyopharm; Honoraria/ad boards: Adaptive, Amgen, Binding Site, BMS, Celgene, Cellectis, Glenmark, Janssen, Juno, Pfizer; and serves on Independent Data Monitoring Committees (IDMCs) for clinical trials lead by Takeda, Merck, Janssen, Theradex. S.Y.K. has received research funding from International Myeloma Foundation, European Research Council, Icelandic Center for Research (Rannís), Amgen, Celgene. The remaining authors declare no competing interests. Funding Information: The iStopMM study is funded by the Black Swan Research Initiative by the International Myeloma Foundation and the Icelandic Centre for Research (grant agreement No 173857). Furthermore, this project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 716677). Screening tests are performed by The Binding Site Ltd. Birmingham, UK. Additional funding is provided by the University of Iceland, Landspítali University Hospital, and the Icelandic Cancer Society. Crosslinking of study data to national registries is performed by the Icelandic Directorate of Health and the Icelandic Cancer Society. The study is made possible by the hundreds of nurses, laboratory technicians, and physicians around Iceland who collect blood samples from participants for screening or during follow-up and provide clinical care that is not part of the study. Icelandic and International myeloma patient organizations including Perluvinir, the Icelandic myeloma patient organization, and the International Myeloma Foundation have provided the project with important perspectives and networks. The information technology department and clinical laboratory staff at LUH made the complex sampling processes required for the study possible. The staff at Loftfar, Aton, Miðlun and Hvíta Húsið played an integral part in the development of recruiting strategies and media campaigns. Decode genetics have generously provided the study team with important insights and access to their state-of-the-art facilities. Presidents Vigdís Finnbogadóttir and Guðni Th Jóhannesson are thanked for their public support of the study. The iStopMM team including Sigurlína H Steinarsdóttir, Hera Hallbera Björnsdóttir, Tinna Hallsdóttir, Ingibjörg Björnsdóttir, and Eva Brynjarsdóttir are particularly acknowledged for their hard and dedicated work. Special thanks go to the residents of Akranes, Iceland who participated in the pilot and proof-of-concept of the study’s recruitment. Most importantly, acknowledgments go to the thousands of Icelanders who have generously provided their informed consent for the study, answered questionnaires, provided blood samples, and undergone diagnostic testing and follow-up. Publisher Copyright: © 2021, The Author(s).Monoclonal gammopathy of undetermined significance (MGUS) precedes multiple myeloma (MM). Population-based screening for MGUS could identify candidates for early treatment in MM. Here we describe the Iceland Screens, Treats, or Prevents Multiple Myeloma study (iStopMM), the first population-based screening study for MGUS including a randomized trial of follow-up strategies. Icelandic residents born before 1976 were offered participation. Blood samples are collected alongside blood sampling in the Icelandic healthcare system. Participants with MGUS are randomized to three study arms. Arm 1 is not contacted, arm 2 follows current guidelines, and arm 3 follows a more intensive strategy. Participants who progress are offered early treatment. Samples are collected longitudinally from arms 2 and 3 for the study biobank. All participants repeatedly answer questionnaires on various exposures and outcomes including quality of life and psychiatric health. National registries on health are cross-linked to all participants. Of the 148,704 individuals in the target population, 80 759 (54.3%) provided informed consent for participation. With a very high participation rate, the data from the iStopMM study will answer important questions on MGUS, including potentials harms and benefits of screening. The study can lead to a paradigm shift in MM therapy towards screening and early therapy.Peer reviewe