Assessing average somatic CAG repeat instability at the protein level

被引：19

作者：

Aviolat, Hubert ^{[1
]}

Pinto, Ricardo Mouro ^{[2
]}

Godschall, Elizabeth ^{[1
]}

Murtha, Ryan ^{[2
]}

Richey, Hannah E. ^{[1
]}

Sapp, Ellen ^{[1
]}

Vodicka, Petr ^{[1
,3
]}

Wheeler, Vanessa C. ^{[2
]}

Kegel-Gleason, Kimberly B. ^{[1
]}

DiFiglia, Marian ^{[1
]}

机构：

[1] Harvard Med Sch, Massachusetts Gen Hosp, MassGen Inst Neurodegenerat Dis, Dept Neurol, Boston, MA 02115 USA

[2] Harvard Med Sch, Ctr Genom Med, Massachusetts Gen Hosp, Dept Neurol, Boston, MA 02115 USA

[3] Czech Acad Sci, Res Ctr PIGMOD, Inst Anim Physiol & Genet, Libechov, Czech Republic

来源：

SCIENTIFIC REPORTS | 2019年 / 9卷 / 1期

关键词：

HUNTINGTONS-DISEASE; MUTANT HUNTINGTIN; TRINUCLEOTIDE REPEAT; CEREBROSPINAL-FLUID; WILD-TYPE; POLYGLUTAMINE; EXPANSION; ONSET; BRAIN; AGE;

D O I：

10.1038/s41598-019-55202-x

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Sandwich ELISA-based methods use Abs that target the expanded polyglutamine (polyQ) tract to quantify mutant huntingtin (mHTT). Using Meso Scale Discovery (MSD) assay, the mHTT signal detected with MW1Ab correlated with polyQ length and doubled with a difference of only 7 glutamine residues between equivalent amounts of purified mHTTexon1 proteins. Similar polyQ length-dependent effects on MSD signals were confirmed using endogenous full length mHTT from brains of Huntington's disease (HD) knock-in (KI) mice. We used this avidity bias to devise a method to assess average CAG repeat instability at the protein level in a mixed population of HTT proteins present in tissues. Signal detected for average polyQ length quantification at the protein level by our method exhibited a strong correlation with average CAG repeat length at the genomic DNA level determined by PCR method in striatal tissue homogenates from Hdh(Q140) KI mice and in human HD postmortem cortex. This work establishes that CAG repeat instability in mutant HTT is reflected at the protein level.

引用

页数：14

共 67 条

[1] CAG EXPANSION AFFECTS THE EXPRESSION OF MUTANT HUNTINGTIN IN THE HUNTINGTONS-DISEASE BRAIN [J].