Testing for mTOR activity

Low-dose rapamycin is recommended to reduce excessive mTOR activity in aging individuals.  However, I have not seen a recommended optimal range of mTOR activity or any testing to establish individual mTOR activity.  I came across an abstract for a testing procedure for high-dose rapamycin use (organ transplant anti-rejection) copied below.  Is there a possibility that this test protocol could be used to find our baseline mTOR activity and establish an optimal mTOR activity level?

PubMed

US National Library of Medicine National Institutes of Health

Journal Biochem. 2010 Aug;403(1-2):79-87. doi: 10.1016/j.ab.2010.04.022. Epub 2010 Apr 24.

A spectrophotometric assay for routine measurement of mammalian target of rapamycin activity in cell lysates.

Dekter HE1, Romijn FP, Temmink WP, van Pelt J, de Fijter JW, Smit NP.

Author information

Abstract

The mammalian target of rapamycin (mTOR) is an important mediator in the PI3K/AKT signaling pathway. mTOR is the target of immunosuppressive drugs, such as rapamycin and everolimus, that are used in transplant patients but also for the treatment of various cancers. We have developed a method for mTOR activity measurement in cell lysates that measures the phosphorylation of p70 S6 kinase by an enzyme linked immunosorbent assay (ELISA) protocol. Using an optimized lysis composition, activity could be measured in the peripheral blood mononuclear cells (PBMCs) isolated from blood. For the PBMCs, intra- and interassay variations of 7 and 10%, respectively, were found using one lot number of the kit. With different lot numbers, the interassay variation increased up to 21%. Activity remained constant for PBMC pool samples on storage for a period of more than 7 months. Activity could also be measured in CD3+ T-cells isolated from blood. In vitro experiments revealed maximum mTOR inhibition of 30% in PBMCs and 44% in T-cells. The in vitro inhibition in PBMCs could also be demonstrated by Western blotting. The mTOR activity measurements may be used to show in vivo inhibition in renal allograft patients during everolimus treatment and to study mTOR activity in various (tumor) cell types.

 

PMID:

20417611

DOI:

10.1016/j.ab.2010.04.022

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