Haemophilia (2014), 20, 304–305
DOI: 10.1111/hae.12418
COMMENTARY
Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS): peering into the cup of Jamshid L. A. VALENTINO Hemophilia and Thrombophilia Center RUSH University Medical Center, Chicago, IL, USA
Summary. The complex process underlying the development of blood-induced joint disease remains mysterious. Novel technologies such as matrix-assisted laser desorption/ionization (MALDI) imaging mass
spectrometry (IMS) to examine protein signatures may provide clues into the process.
Haemophilia is characterized by bleeding due to the absence or diminution of coagulation factor (F) VIII or FIX. In severe cases (residual circulating factor levels of less than 1 IU dL 1), bleeding into the joints is frequent and results in blood-induced joint damage or haemophilic arthropathy (HA) [1]. Currently, the diagnosis of HA is hampered by crude clinical assessments and/or expensive and time-consuming imaging techniques. Furthermore, detection of the earliest stages of HA, when joint damage is potentially reversible – is not yet a reality. Our understanding of what ensues when blood enters the joint space remains an open question, but the pathobiological mechanisms are surely complex. Experimental evidence suggests that danger signals [2] released following tissue damage and erythrocyte lysis, [3] so-called danger-associated molecular patterns (DAMPs), interact with pattern recognition receptors in molecular platforms or inflammasomes located on the surface of classic (mobile) immune cells (i.e. dendritic cells, macrophages, neutrophils) and with stationary, non-immune cells (i.e. vascular cells, epithelial cells, fibroblasts) leading to auto-activation of caspase 1. Caspase 1 in turn cleaves the precursors of cytokines, such as interleukin (IL) 1b, IL-6, and IL-18 along with growth factors (e.g. granulocyte colony stimulating factor, vascular endothelial growth factor), prompting proliferation of synovial fibroblasts and angioblasts that cause the synovial membrane to
become hypertrophic and hypervascular. Iron from degraded erythrocytes, haemosiderin to which iron is bound, and secreted cytokines further activate the immune system through nuclear factor kappa-lightchain-enhancer of activated B cells [4] and the generation of hydroxyl free radicals, the latter having a direct toxic effect leading to chondrocyte apoptosis [5]. Cartilage degradation may be further mediated by production of matrix metalloproteinases, collagenases and plasmin, as is the case in osteoarthritis [6]. Underlying bone metabolism is likely affected by the local cytokine storm progressing within the joint similar to that described in experimental arthritis [7], resulting in osteoporosis and osteophyte and cyst formation. It is probable that one or more likely a panel of these proteins can be identified, the expression of which is linked to the future development of HA. Identification of such biomarkers at an early, reversible stage in the pathobiological development of HA would amount to finding the elusive Holy Grail of haemophilia. In this edition of Haemophilia, Kriegsman and colleagues used matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) to examine protein signatures in the synovial tissue from six haemophilia patients. MALDI IMS has the potential to identify hundreds or possibly thousands of proteins in tissue sections, thereby combining the power of proteomics with morphological assessment. The authors identified signals consistent with ferritin light and heavy chains, alpha- and beta haemoglobin subunits, truncated coagulation FVIII peptide, beta- and gamma fibrinogen peptides and annexin A2. The application of proteomics has the advantage of not requiring a priori knowledge of the proteins of interest, facilitating biomarker discovery. MALDI can be considered a type of multiplex assay that affords the opportunity to assess
Correspondence: Leonard A. Valentino, Hemophilia and Thrombophilia Center, RUSH University Medical Center, Chicago, IL 60612-3833, USA. Tel.: +1 312 942 8114; fax: +1 312 942 8975; e-mail: [email protected] Accepted after revision 24 February 2014 304
Keywords: biomarkers, haemophilia, joint disease, synovitis
© 2014 John Wiley & Sons Ltd
COMMENTARY
many targets in a single assay while providing a spatial orientation of the targets to one another and to other structures within the tissue of interest. Application of this technology to the study of haemophilia and HA may provide a peek into the Cup of Jamshid, which in ancient times was thought to reflect all seven heavens of the universe. Divinations within the cup were considered to reveal deep truths. If MALDI IMS will similarly reveal the truths underlying blood-induced joint disease in haemophilia remains to be determined.
References 1 Valentino LA. Blood-induced joint disease: the pathophysiology of hemophilic arthropathy. J Thromb Haemost 2010; 8: 1895–902. 2 Matzinger P. Immunology. Memories are made of this? Nature 1994; 369: 605–6. 3 Land WG. Transfusion-related acute lung injury: the work of DAMPs. Transfus Med Hemother 2013; 40: 3–13.
© 2014 John Wiley & Sons Ltd
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Disclosures Rush University Medical Center (RUMC) received grant support on behalf of LAV from Baxter Bioscience, Bayer Healthcare, Biogen, CSL Behring, GTC Biotherapeutics, Inspiration Bioscience, NovoNordisk, and Pfizer; and RUMC also received payments on behalf of LAV for his participation in Advisory Boards and as a consultant for Baxter Bioscience, Bayer Healthcare, Biogen, CSL Behring, GTC Biotherapeutics, rEVO Biologics, Inspiration Bioscience, NovoNordisk, and Pfizer. Since the preparation of this manuscript, LAV became an employee of Baxter Bioscience but continues in an academic capacity as a faculty member of RUMC.
4 Sen D, Chapla A, Walter N, Daniel V, Srivastava A, Jayandharan GR. Nuclear factor (NF)-kappaB and its associated pathways are major molecular regulators of blood-induced joint damage in a murine model of hemophilia. J Thromb Haemost 2013; 11: 293– 306. 5 Hooiveld M, Roosendaal G, Wenting M, van den Berg M, Bijlsma J, Lafeber F. Shortterm exposure of cartilage to blood results in
chondrocyte apoptosis. Am J Pathol 2003; 162: 943–51. 6 Fernandes JC, Martel-Pelletier J, Pelletier JP. The role of cytokines in osteoarthritis pathophysiology. Biorheology 2002; 39: 237–46. 7 Kawane K, Tanaka H, Kitahara Y, Shimaoka S, Nagata S. Cytokine-dependent but acquired immunity-independent arthritis caused by DNA escaped from degradation. Proc Natl Acad Sci U S A 2010; 107: 19432–7.
Haemophilia (2014), 20, 304--305
DOI: 10.1111/hae.12418
COMMENTARY
Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS): peering into the cup of Jamshid L. A. VALENTINO Hemophilia and Thrombophilia Center RUSH University Medical Center, Chicago, IL, USA
Summary. The complex process underlying the development of blood-induced joint disease remains mysterious. Novel technologies such as matrix-assisted laser desorption/ionization (MALDI) imaging mass
spectrometry (IMS) to examine protein signatures may provide clues into the process.
Haemophilia is characterized by bleeding due to the absence or diminution of coagulation factor (F) VIII or FIX. In severe cases (residual circulating factor levels of less than 1 IU dL 1), bleeding into the joints is frequent and results in blood-induced joint damage or haemophilic arthropathy (HA) [1]. Currently, the diagnosis of HA is hampered by crude clinical assessments and/or expensive and time-consuming imaging techniques. Furthermore, detection of the earliest stages of HA, when joint damage is potentially reversible – is not yet a reality. Our understanding of what ensues when blood enters the joint space remains an open question, but the pathobiological mechanisms are surely complex. Experimental evidence suggests that danger signals [2] released following tissue damage and erythrocyte lysis, [3] so-called danger-associated molecular patterns (DAMPs), interact with pattern recognition receptors in molecular platforms or inflammasomes located on the surface of classic (mobile) immune cells (i.e. dendritic cells, macrophages, neutrophils) and with stationary, non-immune cells (i.e. vascular cells, epithelial cells, fibroblasts) leading to auto-activation of caspase 1. Caspase 1 in turn cleaves the precursors of cytokines, such as interleukin (IL) 1b, IL-6, and IL-18 along with growth factors (e.g. granulocyte colony stimulating factor, vascular endothelial growth factor), prompting proliferation of synovial fibroblasts and angioblasts that cause the synovial membrane to
become hypertrophic and hypervascular. Iron from degraded erythrocytes, haemosiderin to which iron is bound, and secreted cytokines further activate the immune system through nuclear factor kappa-lightchain-enhancer of activated B cells [4] and the generation of hydroxyl free radicals, the latter having a direct toxic effect leading to chondrocyte apoptosis [5]. Cartilage degradation may be further mediated by production of matrix metalloproteinases, collagenases and plasmin, as is the case in osteoarthritis [6]. Underlying bone metabolism is likely affected by the local cytokine storm progressing within the joint similar to that described in experimental arthritis [7], resulting in osteoporosis and osteophyte and cyst formation. It is probable that one or more likely a panel of these proteins can be identified, the expression of which is linked to the future development of HA. Identification of such biomarkers at an early, reversible stage in the pathobiological development of HA would amount to finding the elusive Holy Grail of haemophilia. In this edition of Haemophilia, Kriegsman and colleagues used matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) to examine protein signatures in the synovial tissue from six haemophilia patients. MALDI IMS has the potential to identify hundreds or possibly thousands of proteins in tissue sections, thereby combining the power of proteomics with morphological assessment. The authors identified signals consistent with ferritin light and heavy chains, alpha- and beta haemoglobin subunits, truncated coagulation FVIII peptide, beta- and gamma fibrinogen peptides and annexin A2. The application of proteomics has the advantage of not requiring a priori knowledge of the proteins of interest, facilitating biomarker discovery. MALDI can be considered a type of multiplex assay that affords the opportunity to assess
Correspondence: Leonard A. Valentino, Hemophilia and Thrombophilia Center, RUSH University Medical Center, Chicago, IL 60612-3833, USA. Tel.: +1 312 942 8114; fax: +1 312 942 8975; e-mail: [email protected] Accepted after revision 24 February 2014 304
Keywords: biomarkers, haemophilia, joint disease, synovitis
© 2014 John Wiley & Sons Ltd
COMMENTARY
many targets in a single assay while providing a spatial orientation of the targets to one another and to other structures within the tissue of interest. Application of this technology to the study of haemophilia and HA may provide a peek into the Cup of Jamshid, which in ancient times was thought to reflect all seven heavens of the universe. Divinations within the cup were considered to reveal deep truths. If MALDI IMS will similarly reveal the truths underlying blood-induced joint disease in haemophilia remains to be determined.
References 1 Valentino LA. Blood-induced joint disease: the pathophysiology of hemophilic arthropathy. J Thromb Haemost 2010; 8: 1895–902. 2 Matzinger P. Immunology. Memories are made of this? Nature 1994; 369: 605–6. 3 Land WG. Transfusion-related acute lung injury: the work of DAMPs. Transfus Med Hemother 2013; 40: 3–13.
© 2014 John Wiley & Sons Ltd
305
Disclosures Rush University Medical Center (RUMC) received grant support on behalf of LAV from Baxter Bioscience, Bayer Healthcare, Biogen, CSL Behring, GTC Biotherapeutics, Inspiration Bioscience, NovoNordisk, and Pfizer; and RUMC also received payments on behalf of LAV for his participation in Advisory Boards and as a consultant for Baxter Bioscience, Bayer Healthcare, Biogen, CSL Behring, GTC Biotherapeutics, rEVO Biologics, Inspiration Bioscience, NovoNordisk, and Pfizer. Since the preparation of this manuscript, LAV became an employee of Baxter Bioscience but continues in an academic capacity as a faculty member of RUMC.
4 Sen D, Chapla A, Walter N, Daniel V, Srivastava A, Jayandharan GR. Nuclear factor (NF)-kappaB and its associated pathways are major molecular regulators of blood-induced joint damage in a murine model of hemophilia. J Thromb Haemost 2013; 11: 293– 306. 5 Hooiveld M, Roosendaal G, Wenting M, van den Berg M, Bijlsma J, Lafeber F. Shortterm exposure of cartilage to blood results in
chondrocyte apoptosis. Am J Pathol 2003; 162: 943–51. 6 Fernandes JC, Martel-Pelletier J, Pelletier JP. The role of cytokines in osteoarthritis pathophysiology. Biorheology 2002; 39: 237–46. 7 Kawane K, Tanaka H, Kitahara Y, Shimaoka S, Nagata S. Cytokine-dependent but acquired immunity-independent arthritis caused by DNA escaped from degradation. Proc Natl Acad Sci U S A 2010; 107: 19432–7.
Haemophilia (2014), 20, 304--305
