Untargeted Metabolite Profiling of Cerebrospinal Fluid Uncovers Biomarkers for Severity of Late Infantile Neuronal Ceroid Lipofuscinosis (CLN2, Batten Disease).

Late infantile neuronal ceroid lipofuscinosis (CLN2 disease) is a rare lysosomal storage disorder caused by a monogenetic deficiency of tripeptidyl peptidase-1 (TPP1). Despite knowledge that lipofuscin is the hallmark disease product, the relevant TPP1 substrate and its role in neuronal physiology/pathology is unknown. We hypothesized that untargeted metabolite profiling of cerebrospinal fluid (CSF) could be used as an effective tool to identify disease-associated metabolic disruptions in CLN2 disease, offering the potential to identify biomarkers that inform on disease severity and progression. Accordingly, a mass spectrometry-based untargeted metabolite profiling approach was employed to differentiate CSF from normal vs. CLN2 deficient individuals. Of 1,433 metabolite features surveyed, 29 linearly correlated with currently employed disease severity scores. With tandem mass spectrometry 8 distinct metabolite identities were structurally confirmed based on retention time and fragmentation pattern matches, vs. standards. These putative CLN2 biomarkers include 7 acetylated species - all attenuated in CLN2 compared to controls. Because acetate is the major bioenergetic fuel for support of mitochondrial respiration, deficient acetylated species in CSF suggests a brain energy defect that may drive neurodegeneration. Targeted analysis of these metabolites in CSF of CLN2 patients offers a powerful new approach for monitoring CLN2 disease progression and response to therapy.

Prolyl oligopeptidase and dipeptidyl peptidase II/dipeptidyl peptidase IV ratio in the cerebrospinal fluid in Parkinson's disease: historical overview and future prospects.

Prolyl oligopeptidase (also named prolyl endopeptidase; PREP) hydrolyzes the Pro-Xaa bonds of biologically active oligopeptides on their carboxyl side. In 1987, we detected PREP activity in human cerebrospinal fluid (CSF) using highly sensitive liquid chromatography-fluorometry with succinyl-Gly-Pro-4-methyl-coumarin amide as a new synthetic substrate, and found a marked decrease in its activity in the cerebrospinal fluid (CSF) from patients with Parkinson's disease (PD) as compared with its level in control patients without neurological diseases. In 2013, Hannula et al. found co-localization of PREP with α-synuclein in the postmortem PD brain. Several recent studies also suggest that the level of PREP in the brain of PD patients may be related to dopamine (DA) cell death via promotion of α-synuclein oligomerization and that inhibitors of PREP may play a neuroprotective role in PD. Although the relationship between another family of prolyl oligopeptidase enzymes, dipeptidyl peptidase II (DPP II) and dipeptidyl peptidase IV (DPP IV), and α-synuclein in the PD brain is not yet clear, we found that the DPP II activity/DPP IV activity ratio in the CSF was significantly increased in PD patients. This review discusses the possibility of PREP as well as the DPP II/DPP IV ratio in the CSF as potential biomarkers of PD.

AAV gene transfer delays disease onset in a TPP1-deficient canine model of the late infantile form of Batten disease.

The most common form of the childhood neurodegenerative disease late infantile neuronal ceroid lipofuscinosis (also called Batten disease) is caused by deficiency of the soluble lysosomal enzyme tripeptidyl peptidase 1 (TPP1) resulting from mutations in the TPP1 gene. We tested whether TPP1 gene transfer to the ependyma, the epithelial lining of the brain ventricular system, in TPP1-deficient dogs would be therapeutically beneficial. A one-time administration of recombinant adeno-associated virus (rAAV) expressing canine TPP1 (rAAV.caTPP1) resulted in high expression of TPP1 predominantly in ependymal cells and secretion of the enzyme into the cerebrospinal fluid leading to clinical benefit. Diseased dogs treated with rAAV.caTPP1 showed delays in onset of clinical signs and disease progression, protection from cognitive decline, and extension of life span. By immunostaining and enzyme assay, recombinant protein was evident throughout the brain and spinal cord, with correction of the neuropathology characteristic of the disease. This study in a naturally occurring canine model of TPP1 deficiency highlights the utility of AAV transduction of ventricular lining cells to accomplish stable secretion of recombinant protein for broad distribution in the central nervous system and therapeutic benefit.

[Activity of DPP III in human cerebrospinal fluid derived from patients with pain].

BACKGROUND: In the course of elucidating physiological roles of spinorphin, an endogenous regulator of enkephalin-degrading enzyme, we found that dipeptidyl peptidase III (DPP III) was contained in human cerebrospinal fluid (CSF). In this study, we examined the activity of DPP III in human CSF derived from patients with pain. METHODS: We used human CSF as specimen collected during operations under spinal anesthesia. Patients were divided into three groups, patients without pain (normal group), with acute pain, and with chronic pain. We measured DPP III activities in each group, using specific substrates, such as Arg-Arg-AMC. RESULTS: DPP III activity for human CSF in patients with acute pain was significantly lower compared with that in patients without pain (P < 0.05). Furthermore, DPP III activity correlated with APN activity, another spinorphin-degrading enzyme, in patients with pain (r = 0.444). CONCLUSION: These results indicate that DPP III may play a role in regulation of endogenous opioids, leading to pain modulation.

[Post-proline hydrolysis activities in cerebrospinal fluid]. / Post-Prolin-Hydrolyseaktivitäten im Liquor cerebrospinalis.

Post-proline hydrolytic-activity exists in various tissues and body fluids of human organism. Using electrophoretic techniques, column chromatographic methods and kinetic investigations two peaks of proteolytic activity towards glycyl-prolyl-p-nitroanilide were detected in human cerebrospinal fluid. The identity of one enzyme with dipeptidylpeptidase IV (EC 3.4.14.5) must now constitute proof of this. Among 650 patients with cerebral diseases, characterised by post-proline hydrolytic activity in cerebrospinal fluid, high enzyme activity coincide with bacterial meningitis. Furthermore, all bacteria which could be isolated from cerebrospinal fluid exhibit high activities of post-proline cleaving hydrolases. Until now, the origin of the cerebrospinal post proline hydrolytic activity is not clear, although our investigations suggested, that lymphocytes, cerebral parenchyma and bacteria may be involved in the enzyme secretion.

Dipeptidyl-aminopeptidase II in human cerebrospinal fluid: changes in patients with Parkinson's disease.

By using a sensitive and specific method, DAP II activity was found in CSF. DAP II activity in CSF of control patients without neurological diseases was 0.416 +/- 0.141 (mean +/- SD) nmole/min/ml and was higher than DAP IV activity in CSF, 0.221 +/- 0.062 (mean +/- SD) nmole/min/ml. In contrast, DAP II activity in serum was 1.16 +/- 0.16 (mean +/- SD) nmole/min/ml and was lower than serum DAP IV activity [41.85 +/- 3.36 (mean +/- SD) nmole/min/ml]. This relatively high activity of DAP II in CSF compared with the activity of DAP IV in CSF together with recent histochemical evidence on the localization of DAP II in some neurons (7) suggests that CSF DAP II may be derived from the brain and may be a marker of some peptidergic neurons. DAP II activity in CSF of patients with Parkinson's disease was significantly increased, whereas DAP IV activity in CSF did not change significantly.

[Validity of post-proline splitting dipeptidyl aminopeptidase activities in the cerebrospinal fluid for neurologic and psychiatric diagnosis]. / Die Validität der postprolinspaltenden Dipeptidyl-aminopeptidase-Aktivitäten im Liquor cerebrospinalis für die neurologisch-psychiatrische Diagnostik.

The dipeptidyl-peptidase activity of 650 liquores cerebrospinalis was obtained by means of the substrate Gly-Pro-NHNp (Gly-Pro-4-nitroanilid) at pHs of 5.5 and 7.4. In comparison with a reference collective, the suitability of the method as a supplementary liquor parameter in neurological psychiatric diagnosis was examined. The introduction of the measurement at pH 7.4 can be particularly recommended for the distinguishing of bacterial and nonbacterial meningitis.

[Determination of dipeptidyl aminopeptidase activities in the cerebrospinal fluid]. / Untersuchungen zur Bestimmung von Dipeptidyl-aminopeptidase-Aktivitäten im Liquor cerebrospinalis.

The investigation of enzyme activity in cerebrospinal fluid has been without relevant results for laboratories analysing spinal fluid. For neurochemical purposes, it is interesting that the Substance P is convered by Depeptidyl-aminopeptidase IV (DP IV), liberating dipeptides. The hydrolysis of nitroanilids of the form Xaa-Pro-NHNp in cerebrospinal fluid was analysed using them as peptidases substrates. Finally a method for measuring the activity was proposed.