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Proteases and protease inhibitors of urinary extracellular vesicles in diabetic nephropathy.

Musante L, Tataruch D, Gu D, Liu X, Forsblom C, Groop PH, Holthofer H - J Diabetes Res (2015)

Bottom Line: Urine samples were divided into groups based on the level of albuminuria and UEVs isolated by hydrostatic dialysis and screened for relative changes of 34 different proteases and 32 protease inhibitors, respectively.Interestingly, myeloblastin and its natural inhibitor elafin showed an increase in the normo- and microalbuminuric groups.Similarly, a characteristic pattern was observed in the array of protease inhibitors, with a marked increase of cystatin B, natural inhibitor of cathepsins L, H, and B as well as of neutrophil gelatinase-associated Lipocalin (NGAL) in the normoalbuminuric group.

View Article: PubMed Central - PubMed

Affiliation: Centre for Bioanalytical Sciences (CBAS), Dublin City University, Dublin 9, Ireland.

ABSTRACT
Diabetic nephropathy (DN) is one of the major complications of diabetes mellitus (DM), leads to chronic kidney disease (CKD), and, ultimately, is the main cause for end-stage kidney disease (ESKD). Beyond urinary albumin, no reliable biomarkers are available for accurate early diagnostics. Urinary extracellular vesicles (UEVs) have recently emerged as an interesting source of diagnostic and prognostic disease biomarkers. Here we used a protease and respective protease inhibitor array to profile urines of type 1 diabetes patients at different stages of kidney involvement. Urine samples were divided into groups based on the level of albuminuria and UEVs isolated by hydrostatic dialysis and screened for relative changes of 34 different proteases and 32 protease inhibitors, respectively. Interestingly, myeloblastin and its natural inhibitor elafin showed an increase in the normo- and microalbuminuric groups. Similarly, a characteristic pattern was observed in the array of protease inhibitors, with a marked increase of cystatin B, natural inhibitor of cathepsins L, H, and B as well as of neutrophil gelatinase-associated Lipocalin (NGAL) in the normoalbuminuric group. This study shows for the first time the distinctive alterations in comprehensive protease profiles of UEVs in diabetic nephropathy and uncovers intriguing mechanistic, prognostic, and diagnostic features of kidney damage in diabetes.

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Related in: MedlinePlus

Urinary and kidney network map. A biological network of interaction using 30 proteases and proteases inhibitor entries extrapolated from the respective arrays was analysed by KUPKB [38]. Selected proteins were 1.5-fold up- or downregulated with respect to the healthy control. According to their intracellular pathways, several proteases can bind and degrade also endogenous inhibitors. This type of binding and interactions can add further complexity to the respective networks, and proteolytic signals themselves may end up in multiple directions. Accordingly, cathepsin L inactivates serpinA1 [75] and MMPs can inactivate a variety of serpins [76]. Conversely, some cystatins interact with MMP-9, still preserving the catalytic function after autodegradation [77]. Interestingly, angiotensinogen (AGT) decreases, through the action of AGT II, the transcription of MMP-1, MMP-2, TIMP-1, TIMP-2, and TIMP-3 but increases MMP-9 in human cardiomyocytes [78].
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fig7: Urinary and kidney network map. A biological network of interaction using 30 proteases and proteases inhibitor entries extrapolated from the respective arrays was analysed by KUPKB [38]. Selected proteins were 1.5-fold up- or downregulated with respect to the healthy control. According to their intracellular pathways, several proteases can bind and degrade also endogenous inhibitors. This type of binding and interactions can add further complexity to the respective networks, and proteolytic signals themselves may end up in multiple directions. Accordingly, cathepsin L inactivates serpinA1 [75] and MMPs can inactivate a variety of serpins [76]. Conversely, some cystatins interact with MMP-9, still preserving the catalytic function after autodegradation [77]. Interestingly, angiotensinogen (AGT) decreases, through the action of AGT II, the transcription of MMP-1, MMP-2, TIMP-1, TIMP-2, and TIMP-3 but increases MMP-9 in human cardiomyocytes [78].

Mentions: In order to establish whether there is any functional interconnection between proteases and protease inhibitors we used the Kidney & Urinary Pathway knowledge Base (KUPKB) [38] designed to collect data set from scientific publications and other datasets related specifically to renal diseases. The query of the KUPKB algorithm provided the protein network presented in Figure 7. Out of 30 entries which are up- or downregulated by 1.5-fold in the protease and protease inhibitor arrays, 11 entities were found to be connected as a network together. Within the network we have found that cathepsin L via cystatin A is connected to the metalloproteases 9 and 2 whose dysregulation is potentially involved in the progression of DN [39]. Interestingly, PRNT-3 mRNA was reported to be upregulated in type 2 diabetic nephropathy [40]. Thus, a further screening in Western blot was carried out evaluating the full cohorts of samples as shown in Figure 8 qualitatively confirming the array data.


Proteases and protease inhibitors of urinary extracellular vesicles in diabetic nephropathy.

Musante L, Tataruch D, Gu D, Liu X, Forsblom C, Groop PH, Holthofer H - J Diabetes Res (2015)

Urinary and kidney network map. A biological network of interaction using 30 proteases and proteases inhibitor entries extrapolated from the respective arrays was analysed by KUPKB [38]. Selected proteins were 1.5-fold up- or downregulated with respect to the healthy control. According to their intracellular pathways, several proteases can bind and degrade also endogenous inhibitors. This type of binding and interactions can add further complexity to the respective networks, and proteolytic signals themselves may end up in multiple directions. Accordingly, cathepsin L inactivates serpinA1 [75] and MMPs can inactivate a variety of serpins [76]. Conversely, some cystatins interact with MMP-9, still preserving the catalytic function after autodegradation [77]. Interestingly, angiotensinogen (AGT) decreases, through the action of AGT II, the transcription of MMP-1, MMP-2, TIMP-1, TIMP-2, and TIMP-3 but increases MMP-9 in human cardiomyocytes [78].
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4383158&req=5

fig7: Urinary and kidney network map. A biological network of interaction using 30 proteases and proteases inhibitor entries extrapolated from the respective arrays was analysed by KUPKB [38]. Selected proteins were 1.5-fold up- or downregulated with respect to the healthy control. According to their intracellular pathways, several proteases can bind and degrade also endogenous inhibitors. This type of binding and interactions can add further complexity to the respective networks, and proteolytic signals themselves may end up in multiple directions. Accordingly, cathepsin L inactivates serpinA1 [75] and MMPs can inactivate a variety of serpins [76]. Conversely, some cystatins interact with MMP-9, still preserving the catalytic function after autodegradation [77]. Interestingly, angiotensinogen (AGT) decreases, through the action of AGT II, the transcription of MMP-1, MMP-2, TIMP-1, TIMP-2, and TIMP-3 but increases MMP-9 in human cardiomyocytes [78].
Mentions: In order to establish whether there is any functional interconnection between proteases and protease inhibitors we used the Kidney & Urinary Pathway knowledge Base (KUPKB) [38] designed to collect data set from scientific publications and other datasets related specifically to renal diseases. The query of the KUPKB algorithm provided the protein network presented in Figure 7. Out of 30 entries which are up- or downregulated by 1.5-fold in the protease and protease inhibitor arrays, 11 entities were found to be connected as a network together. Within the network we have found that cathepsin L via cystatin A is connected to the metalloproteases 9 and 2 whose dysregulation is potentially involved in the progression of DN [39]. Interestingly, PRNT-3 mRNA was reported to be upregulated in type 2 diabetic nephropathy [40]. Thus, a further screening in Western blot was carried out evaluating the full cohorts of samples as shown in Figure 8 qualitatively confirming the array data.

Bottom Line: Urine samples were divided into groups based on the level of albuminuria and UEVs isolated by hydrostatic dialysis and screened for relative changes of 34 different proteases and 32 protease inhibitors, respectively.Interestingly, myeloblastin and its natural inhibitor elafin showed an increase in the normo- and microalbuminuric groups.Similarly, a characteristic pattern was observed in the array of protease inhibitors, with a marked increase of cystatin B, natural inhibitor of cathepsins L, H, and B as well as of neutrophil gelatinase-associated Lipocalin (NGAL) in the normoalbuminuric group.

View Article: PubMed Central - PubMed

Affiliation: Centre for Bioanalytical Sciences (CBAS), Dublin City University, Dublin 9, Ireland.

ABSTRACT
Diabetic nephropathy (DN) is one of the major complications of diabetes mellitus (DM), leads to chronic kidney disease (CKD), and, ultimately, is the main cause for end-stage kidney disease (ESKD). Beyond urinary albumin, no reliable biomarkers are available for accurate early diagnostics. Urinary extracellular vesicles (UEVs) have recently emerged as an interesting source of diagnostic and prognostic disease biomarkers. Here we used a protease and respective protease inhibitor array to profile urines of type 1 diabetes patients at different stages of kidney involvement. Urine samples were divided into groups based on the level of albuminuria and UEVs isolated by hydrostatic dialysis and screened for relative changes of 34 different proteases and 32 protease inhibitors, respectively. Interestingly, myeloblastin and its natural inhibitor elafin showed an increase in the normo- and microalbuminuric groups. Similarly, a characteristic pattern was observed in the array of protease inhibitors, with a marked increase of cystatin B, natural inhibitor of cathepsins L, H, and B as well as of neutrophil gelatinase-associated Lipocalin (NGAL) in the normoalbuminuric group. This study shows for the first time the distinctive alterations in comprehensive protease profiles of UEVs in diabetic nephropathy and uncovers intriguing mechanistic, prognostic, and diagnostic features of kidney damage in diabetes.

Show MeSH
Related in: MedlinePlus