Tau kinetic measurements
The invention relates to in vitro methods for measuring the in vivo metabolism of tau in a subject.
1. An in vitro method for measuring the metabolism of soluble cerebral spinal fluid (CSF) tau in a subject, the method comprising:
(a) administering at least one labeled amino acid to the subject on one or more days;
(b) collecting at least one CSF sample from the subject between day 5 and about day 20, about day 20 and about day 40, about day 40 and about day 100, or a combination thereof;
(c) separating a tau fragment from at least one CSF sample collected in step (b) using an antibody that has affinity within tau's N-terminus or mid-domain;
(d) detecting and measuring by mass spectrometry the amount of labeled tau fragment, or the amount of labeled and unlabeled tau fragment, in at least one sample from step (c); and
(e) calculating the metabolism of soluble CSF tau using the measurements from step (d), wherein the amount of labeled tau fragment in the sample at a given time reflects the metabolism of soluble CSF tau.
2. The method of claim 1 , the method further comprising calculating a metabolic parameter of tau metabolism using the amounts of labeled and/or unlabeled tau determined in claim 1 step (c), the metabolic parameter selected from the group consisting of relative labeling, fractional synthesis rate, fractional clearance rate, absolute synthesis rate, absolute clearance rate, fractional turnover rate, lag time, half-life, peak time, and peak height.
3. An in vitro method for measuring the metabolism of soluble cerebral spinal fluid (CSF) tau in a subject, the method comprising:
(a) separating a tau fragment from at least one CSF sample obtained from a subject, using an antibody that has affinity within tau's N-terminus or mid-domain;
(b) detecting and measuring by mass spectrometry the amount of labeled tau, or the amount of labeled and unlabeled tau, in at least one sample from step (a); and
(b) calculating the metabolism of soluble CSF tau using the measurements from step (b), wherein the amount of labeled tau in the biological sample at a given time reflects the metabolism of soluble CSF tau; and
wherein
(i) the label was administered on one or more days to the subject as one or more labeled amino acids, and (ii) at least one CSF sample was collected from the subject between day 5 and about day 20, day 20 and day 40, day 40 and day 100, or a combination thereof.
4. The method of claim 3 , wherein the labeled amino acid was administered on two or more days between day 0 and about day 3.
5. The method of claim 3 , wherein the labeled amino acid was administered on two or more days between day 0 and about day 5.
6. The method of claim 3 , wherein the labeled amino acid was administered on two or more days between day 0 and about day 10.
7. The method of claim 3 , wherein the labeled amino acid was administered daily.
8. The method of claim 3 , wherein the label is administered to produce an amount of labeled amino acid in the CSF sample selected from the group consisting of about 0.1%, about 0.2%, about 0.5%, about 1%, about 2%, about 5%, about 0.1 to about 20%, and about 0.1 to about 10%.
9. The method of claim 3 , wherein the subject is a rodent.
10. The method of claim 3 , wherein the subject is a human.
11. The method of claim 3 , wherein the tau fragment is separated by immunoprecipitation.
12. The method of claim 3 , the method further comprising calculating a metabolic parameter of soluble tau metabolism using the amounts of labeled and/or unlabeled tau determined in step (b), the metabolic parameter selected from the group consisting of relative labeling, fractional synthesis rate, fractional clearance rate, absolute synthesis rate, absolute clearance rate, fractional turnover rate, lag time, half-life, peak time, and peak height.
13. The method of claim 3 , wherein tau is a phosphorylated tau isoform.
14. The method of claim 3 , wherein at least one CSF sample is collected from the subject between about day 20 and about day 40, about day 40 and about day 100, or a combination thereof.
15. The method of claim 3 , wherein the labeled amino acid is labeled with a non-radioactive isotope.
16. The method of claim 15 , wherein the non-radioactive isotope is selected from the group consisting of 2 H, 13 C, 15 N, 17 O, 18 O, 33 S, 34 S, and 36 S.
17. The method of claim 16 , wherein the labeled amino acid is 13 C 6 leucine.
18. The method of claim 3 , wherein the labeled amino acid was administered to the subject intravenously or orally.
19. The method of claim 18 , wherein the labeled amino acid was administered to the subject orally, at a total daily dose of about 0.1 g to about 10 g, on two or more days between day 0 and about day 10.