PMID- 29641321
OWN - NLM
STAT- MEDLINE
DCOM- 20190520
LR  - 20190802
IS  - 1557-7422 (Electronic)
IS  - 1043-0342 (Print)
IS  - 1043-0342 (Linking)
VI  - 29
IP  - 8
DP  - 2018 Aug
TI  - A Calsequestrin Cis-Regulatory Motif Coupled to a Cardiac Troponin T Promoter 
      Improves Cardiac Adeno-Associated Virus Serotype 9 Transduction Specificity.
PG  - 927-937
LID - 10.1089/hum.2017.188 [doi]
AB  - Adeno-associated virus serotype 9 (AAV9) is an efficient vector for gene transfer 
      to the myocardium. However, the use of ubiquitous promoters, such as the 
      cytomegalovirus (CMV) promoter, can result in expression of the transgene in 
      organs other than the heart. This study tested if the efficiency and specificity 
      of cardiac transcription from a chicken cardiac troponin T (TnT) promoter could 
      be further increased by incorporating a cardiomyocyte-specific transcriptional 
      cis-regulatory motif from human calsequestrin 2 (CS-CRM4) into the expression 
      cassette (Enh.TnT). The efficiency of luciferase expression from the TnT and 
      Enh.TnT constructs was compared to expression of luciferase under the control of 
      the CMV promoter in both adult and neonatal mice. Overall, expression levels of 
      luciferase in the heart were similar in mice injected with AAV9.TnT.Luc, 
      AAV9.Enh.TnT.Luc and AAV9.CMV.Luc. In contrast, expression levels of luciferase 
      activity in nontarget organs, including the liver and muscle, was lower in mice 
      injected with the AAV9.TnT.Luc compared to AAV9.CMV.Luc and was negligible with 
      AAV9.Enh.TnT. In neonates, in organs other than the heart, luciferase expression 
      levels were too low to be quantified for all constructs. Taken together, the data 
      show that the AAV9 Enh.TnT constructs drives high levels of expression of the 
      transgene in the myocardium, with insignificant expression in other organs. These 
      properties reduce the risks associated with the AAV9-mediated expression of the 
      therapeutic protein of interest in nontarget organs. The excellent cardiac 
      specificity should allow for the use of higher vector doses than are currently 
      used, which might be essential to achieve the levels of transgene expression 
      necessary for therapeutic benefits. Taken together, the findings suggest that the 
      Enh.TnT transcription unit is a potentially attractive tool for clinical cardiac 
      gene therapy in adults.
FAU - Chamberlain, Kyle
AU  - Chamberlain K
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - Riyad, Jalish M
AU  - Riyad JM
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - Garnett, Tyrone
AU  - Garnett T
AD  - 3 Center for Cardiovascular Genetics, Institute of Molecular Medicine and 
      Department of Medicine, University of Texas Health Sciences Center at Houston, 
      and Texas Heart Institute, Houston, Texas.
FAU - Kohlbrenner, Erik
AU  - Kohlbrenner E
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - Mookerjee, Ananda
AU  - Mookerjee A
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - Elmastour, Firas
AU  - Elmastour F
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - Benard, Ludovic
AU  - Benard L
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - Chen, Jiqiu
AU  - Chen J
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - VandenDriessche, Thierry
AU  - VandenDriessche T
AD  - 4 Department of Gene Therapy and Regenerative Medicine, Free University of 
      Brussels (VUB), Brussels, Belgium .
AD  - 5 Center for Molecular and Vascular Biology, Department of Cardiovascular 
      Sciences, University of Leuven , Leuven, Belgium .
FAU - Chuah, Marinee K
AU  - Chuah MK
AD  - 4 Department of Gene Therapy and Regenerative Medicine, Free University of 
      Brussels (VUB), Brussels, Belgium .
AD  - 5 Center for Molecular and Vascular Biology, Department of Cardiovascular 
      Sciences, University of Leuven , Leuven, Belgium .
FAU - Marian, Ali J
AU  - Marian AJ
AD  - 3 Center for Cardiovascular Genetics, Institute of Molecular Medicine and 
      Department of Medicine, University of Texas Health Sciences Center at Houston, 
      and Texas Heart Institute, Houston, Texas.
FAU - Hajjar, Roger J
AU  - Hajjar RJ
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
FAU - Gurha, Priyatansh
AU  - Gurha P
AD  - 3 Center for Cardiovascular Genetics, Institute of Molecular Medicine and 
      Department of Medicine, University of Texas Health Sciences Center at Houston, 
      and Texas Heart Institute, Houston, Texas.
FAU - Weber, Thomas
AU  - Weber T
AD  - 1 Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York 
      City, New York.
AD  - 2 Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount 
      Sinai, New York City, New York.
LA  - eng
GR  - R01 HL128099/HL/NHLBI NIH HHS/United States
GR  - R01 HL132401/HL/NHLBI NIH HHS/United States
GR  - R01 HL135093/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20180509
PL  - United States
TA  - Hum Gene Ther
JT  - Human gene therapy
JID - 9008950
RN  - 0 (CASQ2 protein, human)
RN  - 0 (Calsequestrin)
RN  - 0 (Troponin T)
SB  - IM
MH  - Animals
MH  - Animals, Newborn
MH  - Calsequestrin/genetics
MH  - Chickens/genetics
MH  - Dependovirus/*genetics
MH  - Gene Expression Regulation/genetics
MH  - *Genetic Therapy
MH  - Genetic Vectors/genetics/therapeutic use
MH  - Heart Diseases/genetics/*therapy
MH  - Humans
MH  - Mice
MH  - Myocardium/*metabolism/pathology
MH  - Myocytes, Cardiac/metabolism/virology
MH  - Promoter Regions, Genetic/genetics
MH  - *Transduction, Genetic
MH  - Troponin T/genetics
PMC - PMC6098410
OTO - NOTNLM
OT  - AAV9
OT  - CMV
OT  - calsequestrin enhancer
OT  - cardiac tropism
OT  - cardiac-specific cis-regulatory motif
OT  - troponin T
COIS- The authors declare no conflicts of interest.
EDAT- 2018/04/12 06:00
MHDA- 2019/05/21 06:00
PMCR- 2019/08/01
CRDT- 2018/04/12 06:00
PHST- 2018/04/12 06:00 [pubmed]
PHST- 2019/05/21 06:00 [medline]
PHST- 2018/04/12 06:00 [entrez]
PHST- 2019/08/01 00:00 [pmc-release]
AID - 10.1089/hum.2017.188 [pii]
AID - 10.1089/hum.2017.188 [doi]
PST - ppublish
SO  - Hum Gene Ther. 2018 Aug;29(8):927-937. doi: 10.1089/hum.2017.188. Epub 2018 May 
      9.