Amyloid The Journal of Protein Folding Disorders
ISSN: 1350-6129 (Print) 1744-2818 (Online) Journal homepage: http://www.tandfonline.com/loi/iamy20
Hepatocyte growth factor measurement in AL amyloidosis Julie Abraham, Estelle Desport, Charlotte Rigaud, Benoit Marin, Sébastien Bender, Corinne Lacombe, Stéphane Moreau, Fatima Yagoubi, Dominique Bordessoule, David Lavergne, Frank Bridoux & Arnaud Jaccard To cite this article: Julie Abraham, Estelle Desport, Charlotte Rigaud, Benoit Marin, Sébastien Bender, Corinne Lacombe, Stéphane Moreau, Fatima Yagoubi, Dominique Bordessoule, David Lavergne, Frank Bridoux & Arnaud Jaccard (2015) Hepatocyte growth factor measurement in AL amyloidosis, Amyloid, 22:2, 112-116, DOI: 10.3109/13506129.2015.1014548 To link to this article: http://dx.doi.org/10.3109/13506129.2015.1014548
Published online: 08 Jun 2015.
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Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=iamy20 Download by: [Chinese University of Hong Kong]
Date: 08 November 2015, At: 01:40
http://informahealthcare.com/amy ISSN: 1350-6129 (print), 1744-2818 (electronic) Amyloid, 2015; 22(2): 112–116 ! 2015 Informa UK Ltd. DOI: 10.3109/13506129.2015.1014548
ORIGINAL ARTICLE
Hepatocyte growth factor measurement in AL amyloidosis Julie Abraham1, Estelle Desport2, Charlotte Rigaud1, Benoit Marin3, Se´bastien Bender4,5, Corinne Lacombe3, Ste´phane Moreau1, Fatima Yagoubi1, Dominique Bordessoule1, David Lavergne1, Frank Bridoux2,5, and Arnaud Jaccard1,5
Downloaded by [Chinese University of Hong Kong] at 01:40 08 November 2015
1
Service d’He´matologie Clinique, Centre National de Re´fe´rence Amylose AL, CHU, Limoges, France, 2Service de Ne´phrologie, Centre National de Re´fe´rence Amylose AL, CHU, Poitiers, France, 3CHU Limoges, Centre d’Epide´miologie de Biostatistique et de Me´thodologie de la Recherche, France, 4 Service d’Immunologie et d’Immunoge´ne´tique, CHU, Limoges, France, and 5UMR CNRS 7276, Centre National de la Recherche Scientifique, Universite´ de Limoges, Limoges, France Abstract
Keywords
Hepatocyte growth factor (HGF) is a pro-angiogenic cytokine activated by tissue-type plasminogen activator (tPA) that might play a role in the progression of multiple myeloma (MM). Preliminary studies indicated that serum HGF levels were higher in patients with AL amyloidosis (AL) compared to those with MM. The aim of the present study was to determine whether HGF is a relevant marker of diagnosis and prognosis in AL. HGF serum levels were measured at diagnosis in patients with monoclonal gammopathy (MG) without AL (76 controls), or with biopsy-proven systemic AL (69 patients). HGF serum levels were significantly higher in patients with AL compared to controls, respectively, 11.2 ng/mL [min: 0.95–max: 200.4] versus 1.4 ng/mL [min: 0.82–max: 6.2] (p50.0001). The threshold value of 2.2 ng/mL conferred optimal sensitivity (88%) and specificity (95%) to differentiate AL and monoclonal gammopathy of undetermined significance (MGUS) patients. Serum HGF concentrations were correlated positively with the severity of cardiac involvement and the serum level of monoclonal light chains. These data suggest that HGF measurement could be used in patients with MG to detect AL or to reinforce a clinical suspicion of AL and to guide indications for diagnostic tissue biopsies.
Amyloidosis, hepatocyte growth factor, monoclonal gammopathy History Received 16 October 2014 Revised 30 December 2014 Accepted 29 January 2015 Published online 8 June 2015
Abbreviations: HGF: hepatocyte growth factor; LC: immunoglobulin light chain; LVEF: left ventricular ejection fraction; MDRD: modification of diet in renal disease; MG: monoclonal gammopathy; MGUS: monoclonal gammopathy of undetermined significance; MM: multiple myeloma; NT-proBNP: n-terminal pro b-type natriuretic peptide; ROC: receiver operating characteristic; tPA: tissue plasminogen activator
Background Systemic AL amyloidosis is a rare but severe acquired disease in which a monoclonal free light chain deposits as aggregated extracellular fibrils and causes organ dysfunction. The organs most frequently involved are the kidney and the heart. The non-specific and variable symptoms associated with AL frequently lead to such long delays in diagnosis that organ dysfunction, heart dysfunction in particular, is already advanced when treatment is initiated. Hepatocyte growth factor (HGF) is a pro-angiogenic cytokine and a mitogenic, motogenic and morphogenic factors that shares similar structural protein’s organization with plasminogen. It could be activated by tissue-type plasminogen factor (tPA) involved in tumor growth [1].
Previous studies have shown that myeloma cell lines and primary myeloma cells over-express HGF and that elevated HGF levels predict poor prognosis, with short-term responses to therapies and early relapses [2–4]. In a small case series, serum HGF levels were found to be significantly higher in patients with AL amyloidosis compared to patients with multiple myeloma (MM) [3]. A subsequent preliminary study of 18 AA and AL amyloidosis patients [5] suggested that measurement of HGF might be useful for the diagnosis of amyloidosis. To determine whether HGF may be used as a relevant diagnostic and prognostic marker in AL, we compared baseline HGF serum levels at diagnosis in patients with biopsy-proven systemic AL and in patients with monoclonal gammopathies who did not develop systemic AL after a prolonged period of clinical surveillance.
Patients and methods Address for correspondence: Arnaud Jaccard, CHU Limoges, He´matologie Clinique et The´rapie Cellulaire, 2 avenue Martin Luther King, Limoges 87000, France. E-mail: [email protected]
Our study was retrospective and conducted with close collaboration of two French centres.
HGF measurement in AL amyloidosis
Downloaded by [Chinese University of Hong Kong] at 01:40 08 November 2015
DOI: 10.3109/13506129.2015.1014548
The study population consisted in AL amyloidosis patients, aged over 18, seen in these two centres during a five-year period, from whom a serum sample was drawn at diagnosis before introduction of any treatment. The control population was composed of patients with monoclonal gammopathy of undetermined significance (MGUS) and MM. Controls’ serum samples were available at time of diagnosis of the monoclonal gammopathy (MG), and none of the control patients developed symptoms suggestive of systemic AL amyloidosis after a median follow-up of 61 [7–106] months. The diagnosis of amyloidosis was confirmed by biopsy in all cases, based on the demonstration of Congo red-positive tissue deposits, with typical green birefringence under polarized light and specific staining with anti-lambda or anti-kappa LC conjugate by immunohistochemistry or immunofluorescence [6–10]. Diagnosis criteria for MGUS and MM were based on criteria reviewed by Sirohi and Powles [11]. Clinical parameters, such as age, sex and organ involvement in the AL patients were systematically collected. Organ involvement was defined according to the consensus criteria published in 2005 [12] and modified in 2012 [13]. Biological parameters at diagnosis, such as serum creatinine, serum free light chains, troponin T and NT-proBNP levels were systematically recorded. Creatinine clearance was calculated using the modification of diet in renal disease (MDRD) formula. Echographical parameters, such as left ventricular ejection fraction (LVEF) and septal thickness were also collected. AL patients were stratified according to the Mayo clinic staging based on NT-proBNP and troponin T levels, measured with an electrochemiluminescence immunoassay ‘‘CobasÕ’’ (Roche Diagnostics, Baˆle, Switzerland). Serum HGF level was measured by enzyme-linked immunosorbent assay (QuantikineÕ , R&D Systems, Minneapolis, MN), according to the manufacturer’s instructions. The detection limit was 125 pg/mL. HGF level was measured retrospectively on serum samples taken at diagnosis and stored at 20 C or 80 C in 69 patients with AL amyloidosis, 56 patients with MGUS and 17 patients with MM. The study was approved by the local ethics committee (CHU Limoges, France) and written informed consent was obtained from all patients and controls. The primary objective was to evaluate the accuracy of HGF measurement for the diagnosis of systemic AL amyloidosis among patients with MG. The secondary objectives were to determine which organ involvements were associated with high-HGF levels and to evaluate the role of HGF level to predict outcome. Statistical analysis Quantitative variables were presented by median and [min– max] or [p25–p75] interval as clinically relevant and compared using the non-parametric Wilcoxon test. Qualitative variables were described by number and percentage and differences were compared using Fisher’s exact test for categorical variables when appropriate. Receiver operating characteristic (ROC) curve was used to search specific and sensitive value of HGF aiming to identify AL amyloidosis
113
patients among patients with gammopathies. Analyses were done using the group with MGUS as control group and HGF was also measured in the group with MM. The association between organ involvement and serum HGF level was studied using the consensus criteria [13,14] to define organ involvement and using the creatinine clearance. Correlation between serum HGF level and NT-proBNP, troponin T, creatinine clearance was studied using the Spearman correlation coefficient. Survival analysis using patients with AL amyloidosis according to their initial levels of HGF was performed using the Kaplan–Meier and log-rank for comparisons among groups. Cox proportional hazard model was performed to evaluate relative risks between groups.
Results Patients’ characteristics Sixty-nine AL patients, median age 61 [min: 32–max: 90], were included in two French reference centres. Clinical and biological characteristics are summarized in Table 1. Control group included 56 patients with MGUS and 17 patients with MM without evidence of AL. Serum creatinine levels were slightly different (p ¼ 0.02); with, respectively, 88 mmol/L [min: 39–max: 751] in AL patients versus 76 mmol/L (48–137) in patients with MG. Sixteen AL patients (24%) had a creatinine clearance below 50 mL/min. HGF measurement accuracy Median serum HGF levels were significantly higher in AL patients compared to MGUS: 11.2 ng/mL [min: 0.96–max: 200.4] versus 1.4 ng/mL [min: 0.82–max: 6.2] (p50.0001). Moreover, using ROC curve, HGF levels appeared as a discriminating factor for the diagnosis of AL amyloidosis compared to MGUS patients with an area under curve of 0.93 (CI 95% [0.88–0.98]) (p ¼ 0.0002). The threshold value
Table 1. Clinical and biological characteristics of AL patients. n (%) Men/Women Age (Years) Organ involvement Kidney Proteinuria (g/24H) Heart Liver Soft tissue Nerve Gut Isotype (mg/L) Lambda Kappa Abnormal FLC Severity stage* I II III NT-proBNP (ng/L) Troponin T (mg/L)
Median (min–max)
46/23 61 (32–90) 57 (82.6) 3.19 (0.2–40.8) 40 27 23 18 12
(57.9) (39.1) (33.3) (26.1) (17.4)
48 (69.6) 21 (30.4)
204 (15–2580) 157.5 (16–2620) 182 (15–2580)
25 (36.2) 13 (18.8) 29 (42) 862 (9–125 283) 0.028 (0.01–1.26)
*According to Mayo Clinic staging, NT-proBNP 5 or 332 ng/mL, troponin T 5 or 0.035 mg/L.
log HGF
(A)
J. Abraham et al.
Amyloid, 2015; 22(2): 112–116
(B)
P
ISSN: 1350-6129 (Print) 1744-2818 (Online) Journal homepage: http://www.tandfonline.com/loi/iamy20
Hepatocyte growth factor measurement in AL amyloidosis Julie Abraham, Estelle Desport, Charlotte Rigaud, Benoit Marin, Sébastien Bender, Corinne Lacombe, Stéphane Moreau, Fatima Yagoubi, Dominique Bordessoule, David Lavergne, Frank Bridoux & Arnaud Jaccard To cite this article: Julie Abraham, Estelle Desport, Charlotte Rigaud, Benoit Marin, Sébastien Bender, Corinne Lacombe, Stéphane Moreau, Fatima Yagoubi, Dominique Bordessoule, David Lavergne, Frank Bridoux & Arnaud Jaccard (2015) Hepatocyte growth factor measurement in AL amyloidosis, Amyloid, 22:2, 112-116, DOI: 10.3109/13506129.2015.1014548 To link to this article: http://dx.doi.org/10.3109/13506129.2015.1014548
Published online: 08 Jun 2015.
Submit your article to this journal
Article views: 41
View related articles
View Crossmark data
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=iamy20 Download by: [Chinese University of Hong Kong]
Date: 08 November 2015, At: 01:40
http://informahealthcare.com/amy ISSN: 1350-6129 (print), 1744-2818 (electronic) Amyloid, 2015; 22(2): 112–116 ! 2015 Informa UK Ltd. DOI: 10.3109/13506129.2015.1014548
ORIGINAL ARTICLE
Hepatocyte growth factor measurement in AL amyloidosis Julie Abraham1, Estelle Desport2, Charlotte Rigaud1, Benoit Marin3, Se´bastien Bender4,5, Corinne Lacombe3, Ste´phane Moreau1, Fatima Yagoubi1, Dominique Bordessoule1, David Lavergne1, Frank Bridoux2,5, and Arnaud Jaccard1,5
Downloaded by [Chinese University of Hong Kong] at 01:40 08 November 2015
1
Service d’He´matologie Clinique, Centre National de Re´fe´rence Amylose AL, CHU, Limoges, France, 2Service de Ne´phrologie, Centre National de Re´fe´rence Amylose AL, CHU, Poitiers, France, 3CHU Limoges, Centre d’Epide´miologie de Biostatistique et de Me´thodologie de la Recherche, France, 4 Service d’Immunologie et d’Immunoge´ne´tique, CHU, Limoges, France, and 5UMR CNRS 7276, Centre National de la Recherche Scientifique, Universite´ de Limoges, Limoges, France Abstract
Keywords
Hepatocyte growth factor (HGF) is a pro-angiogenic cytokine activated by tissue-type plasminogen activator (tPA) that might play a role in the progression of multiple myeloma (MM). Preliminary studies indicated that serum HGF levels were higher in patients with AL amyloidosis (AL) compared to those with MM. The aim of the present study was to determine whether HGF is a relevant marker of diagnosis and prognosis in AL. HGF serum levels were measured at diagnosis in patients with monoclonal gammopathy (MG) without AL (76 controls), or with biopsy-proven systemic AL (69 patients). HGF serum levels were significantly higher in patients with AL compared to controls, respectively, 11.2 ng/mL [min: 0.95–max: 200.4] versus 1.4 ng/mL [min: 0.82–max: 6.2] (p50.0001). The threshold value of 2.2 ng/mL conferred optimal sensitivity (88%) and specificity (95%) to differentiate AL and monoclonal gammopathy of undetermined significance (MGUS) patients. Serum HGF concentrations were correlated positively with the severity of cardiac involvement and the serum level of monoclonal light chains. These data suggest that HGF measurement could be used in patients with MG to detect AL or to reinforce a clinical suspicion of AL and to guide indications for diagnostic tissue biopsies.
Amyloidosis, hepatocyte growth factor, monoclonal gammopathy History Received 16 October 2014 Revised 30 December 2014 Accepted 29 January 2015 Published online 8 June 2015
Abbreviations: HGF: hepatocyte growth factor; LC: immunoglobulin light chain; LVEF: left ventricular ejection fraction; MDRD: modification of diet in renal disease; MG: monoclonal gammopathy; MGUS: monoclonal gammopathy of undetermined significance; MM: multiple myeloma; NT-proBNP: n-terminal pro b-type natriuretic peptide; ROC: receiver operating characteristic; tPA: tissue plasminogen activator
Background Systemic AL amyloidosis is a rare but severe acquired disease in which a monoclonal free light chain deposits as aggregated extracellular fibrils and causes organ dysfunction. The organs most frequently involved are the kidney and the heart. The non-specific and variable symptoms associated with AL frequently lead to such long delays in diagnosis that organ dysfunction, heart dysfunction in particular, is already advanced when treatment is initiated. Hepatocyte growth factor (HGF) is a pro-angiogenic cytokine and a mitogenic, motogenic and morphogenic factors that shares similar structural protein’s organization with plasminogen. It could be activated by tissue-type plasminogen factor (tPA) involved in tumor growth [1].
Previous studies have shown that myeloma cell lines and primary myeloma cells over-express HGF and that elevated HGF levels predict poor prognosis, with short-term responses to therapies and early relapses [2–4]. In a small case series, serum HGF levels were found to be significantly higher in patients with AL amyloidosis compared to patients with multiple myeloma (MM) [3]. A subsequent preliminary study of 18 AA and AL amyloidosis patients [5] suggested that measurement of HGF might be useful for the diagnosis of amyloidosis. To determine whether HGF may be used as a relevant diagnostic and prognostic marker in AL, we compared baseline HGF serum levels at diagnosis in patients with biopsy-proven systemic AL and in patients with monoclonal gammopathies who did not develop systemic AL after a prolonged period of clinical surveillance.
Patients and methods Address for correspondence: Arnaud Jaccard, CHU Limoges, He´matologie Clinique et The´rapie Cellulaire, 2 avenue Martin Luther King, Limoges 87000, France. E-mail: [email protected]
Our study was retrospective and conducted with close collaboration of two French centres.
HGF measurement in AL amyloidosis
Downloaded by [Chinese University of Hong Kong] at 01:40 08 November 2015
DOI: 10.3109/13506129.2015.1014548
The study population consisted in AL amyloidosis patients, aged over 18, seen in these two centres during a five-year period, from whom a serum sample was drawn at diagnosis before introduction of any treatment. The control population was composed of patients with monoclonal gammopathy of undetermined significance (MGUS) and MM. Controls’ serum samples were available at time of diagnosis of the monoclonal gammopathy (MG), and none of the control patients developed symptoms suggestive of systemic AL amyloidosis after a median follow-up of 61 [7–106] months. The diagnosis of amyloidosis was confirmed by biopsy in all cases, based on the demonstration of Congo red-positive tissue deposits, with typical green birefringence under polarized light and specific staining with anti-lambda or anti-kappa LC conjugate by immunohistochemistry or immunofluorescence [6–10]. Diagnosis criteria for MGUS and MM were based on criteria reviewed by Sirohi and Powles [11]. Clinical parameters, such as age, sex and organ involvement in the AL patients were systematically collected. Organ involvement was defined according to the consensus criteria published in 2005 [12] and modified in 2012 [13]. Biological parameters at diagnosis, such as serum creatinine, serum free light chains, troponin T and NT-proBNP levels were systematically recorded. Creatinine clearance was calculated using the modification of diet in renal disease (MDRD) formula. Echographical parameters, such as left ventricular ejection fraction (LVEF) and septal thickness were also collected. AL patients were stratified according to the Mayo clinic staging based on NT-proBNP and troponin T levels, measured with an electrochemiluminescence immunoassay ‘‘CobasÕ’’ (Roche Diagnostics, Baˆle, Switzerland). Serum HGF level was measured by enzyme-linked immunosorbent assay (QuantikineÕ , R&D Systems, Minneapolis, MN), according to the manufacturer’s instructions. The detection limit was 125 pg/mL. HGF level was measured retrospectively on serum samples taken at diagnosis and stored at 20 C or 80 C in 69 patients with AL amyloidosis, 56 patients with MGUS and 17 patients with MM. The study was approved by the local ethics committee (CHU Limoges, France) and written informed consent was obtained from all patients and controls. The primary objective was to evaluate the accuracy of HGF measurement for the diagnosis of systemic AL amyloidosis among patients with MG. The secondary objectives were to determine which organ involvements were associated with high-HGF levels and to evaluate the role of HGF level to predict outcome. Statistical analysis Quantitative variables were presented by median and [min– max] or [p25–p75] interval as clinically relevant and compared using the non-parametric Wilcoxon test. Qualitative variables were described by number and percentage and differences were compared using Fisher’s exact test for categorical variables when appropriate. Receiver operating characteristic (ROC) curve was used to search specific and sensitive value of HGF aiming to identify AL amyloidosis
113
patients among patients with gammopathies. Analyses were done using the group with MGUS as control group and HGF was also measured in the group with MM. The association between organ involvement and serum HGF level was studied using the consensus criteria [13,14] to define organ involvement and using the creatinine clearance. Correlation between serum HGF level and NT-proBNP, troponin T, creatinine clearance was studied using the Spearman correlation coefficient. Survival analysis using patients with AL amyloidosis according to their initial levels of HGF was performed using the Kaplan–Meier and log-rank for comparisons among groups. Cox proportional hazard model was performed to evaluate relative risks between groups.
Results Patients’ characteristics Sixty-nine AL patients, median age 61 [min: 32–max: 90], were included in two French reference centres. Clinical and biological characteristics are summarized in Table 1. Control group included 56 patients with MGUS and 17 patients with MM without evidence of AL. Serum creatinine levels were slightly different (p ¼ 0.02); with, respectively, 88 mmol/L [min: 39–max: 751] in AL patients versus 76 mmol/L (48–137) in patients with MG. Sixteen AL patients (24%) had a creatinine clearance below 50 mL/min. HGF measurement accuracy Median serum HGF levels were significantly higher in AL patients compared to MGUS: 11.2 ng/mL [min: 0.96–max: 200.4] versus 1.4 ng/mL [min: 0.82–max: 6.2] (p50.0001). Moreover, using ROC curve, HGF levels appeared as a discriminating factor for the diagnosis of AL amyloidosis compared to MGUS patients with an area under curve of 0.93 (CI 95% [0.88–0.98]) (p ¼ 0.0002). The threshold value
Table 1. Clinical and biological characteristics of AL patients. n (%) Men/Women Age (Years) Organ involvement Kidney Proteinuria (g/24H) Heart Liver Soft tissue Nerve Gut Isotype (mg/L) Lambda Kappa Abnormal FLC Severity stage* I II III NT-proBNP (ng/L) Troponin T (mg/L)
Median (min–max)
46/23 61 (32–90) 57 (82.6) 3.19 (0.2–40.8) 40 27 23 18 12
(57.9) (39.1) (33.3) (26.1) (17.4)
48 (69.6) 21 (30.4)
204 (15–2580) 157.5 (16–2620) 182 (15–2580)
25 (36.2) 13 (18.8) 29 (42) 862 (9–125 283) 0.028 (0.01–1.26)
*According to Mayo Clinic staging, NT-proBNP 5 or 332 ng/mL, troponin T 5 or 0.035 mg/L.
log HGF
(A)
J. Abraham et al.
Amyloid, 2015; 22(2): 112–116
(B)
P
