A Novel mouse model of enhanced proteostasis: Full-length human heat shock factor 1 transgenic mice.

The heat shock response (HSR) is controlled by the master transcriptional regulator heat shock factor 1 (HSF1). HSF1 maintains proteostasis and resistance to stress through production of heat shock proteins (HSPs). No transgenic model exists that overexpresses HSF1 in tissues of the central nervous system (CNS). We generated a transgenic mouse overexpressing full-length non-mutant HSF1 and observed a 2-4-fold increase in HSF1 mRNA and protein expression in all tissues studied of HSF1 transgenic (HSF1(+/0)) mice compared to wild type (WT) littermates, including several regions of the CNS. Basal expression of HSP70 and 90 showed only mild tissue-specific changes; however, in response to forced exercise, the skeletal muscle HSR was more elevated in HSF1(+/0) mice compared to WT littermates and in fibroblasts following heat shock, as indicated by levels of inducible HSP70 mRNA and protein. HSF1(+/0) cells elicited a significantly more robust HSR in response to expression of the 82 repeat polyglutamine-YFP fusion construct (Q82YFP) and maintained proteasome-dependent processing of Q82YFP compared to WT fibroblasts. Overexpression of HSF1 was associated with fewer, but larger Q82YFP aggregates resembling aggresomes in HSF1(+/0) cells, and increased viability. Therefore, our data demonstrate that tissues and cells from mice overexpressing full-length non-mutant HSF1 exhibit enhanced proteostasis.

Plasma protein biomarkers of the geriatric syndrome of frailty.

Frailty is a geriatric syndrome associated with physical decline with aging. Using a proteomics-based screening method to screen plasma for potential biomarkers, we previously found inflammatory glycoproteins to be increased with frailty. The purpose of this study was to confirm if plasma levels of these glycoproteins, as well as of interleukin-6, are increased with frailty in a larger sample (n = 65) of community-dwelling older adults. Plasma levels of transferrin, fibrinogen, haptoglobin, and interleukin-6 were determined with enzyme-linked immunosorbent assay. Differences in protein concentrations by frailty status were determined using analysis of variance. Higher levels of transferrin (p < .001), fibrinogen (p < .0001), and interleukin-6 (p = .0035) were associated with frailty status (nonfrail, prefrail, or frail) and frailty score (0-5) in this sample even after adjustment for age and sex. Haptoglobin did not differ by frailty status (p = .05). Our findings largely confirmed the findings of our nontargeted approach that inflammatory glycoproteins are increased with frailty. Future studies should include larger examinations of these associations and consider the potential usefulness of these glycoproteins as biomarkers for frailty.

Proteomic screening of glycoproteins in human plasma for frailty biomarkers.

The application of proteomics methodology for analyzing human blood samples is of increasing importance as a noninvasive method for understanding, detecting, and monitoring disease. In particular, glycoproteomic analysis may be useful in the study of age-related diseases and syndromes, such as frailty. This study demonstrates the use of methodology for isolating plasma glycoproteins using lectins, comparing the glycoproteome by frailty status using two-dimensional polyacrylamide gel electrophoresis and identifying glycoproteins using mass spectrometry. In a pilot study, we found seven glycoproteins to differ by at least twofold in prefrail compared with nonfrail older adults, including haptoglobin, transferrin, and fibrinogen, consistent with known inflammatory and hematologic changes associated with frailty. Enzyme-linked immunosorbent assay analysis found that plasma transferrin concentration was increased in frail and prefrail older adults compared with nonfrail, confirming our proteomic findings. This work provides evidence for using a reproducible methodology for conducting clinical proteomic comparative studies of age-related diseases.