CN111154757B - 3' -UTR sequence of Aggf1 gene and application thereof - Google Patents

Abstract

The invention relates to a nucleic acid element capable of regulating gene expression in cells, which is a 3' -UTR sequence of human or mouse Aggf 1; the application of the nucleic acid element in preparing heart reconstruction therapeutic drugs and the application of the nucleic acid element in regulating gene expression in isolated cells are also disclosed; the invention also provides application of the nucleic acid construct in preparing a gene therapy drug; also relates to a nucleic acid construct comprising the nucleic acid element and application thereof. For example, AAV delivery systems can be used to deliver such nucleic acid constructs into cells, or to foci, to modulate expression of related genes, such as Aggf1 and Mcl1, in related cells or tissues, for therapeutic purposes. The nucleic acid element of the invention can be introduced into damaged heart tissue in series with the open reading frame of the Aggf1 gene, which has better therapeutic effect than just introducing the open reading frame.

Description

3' -UTR sequence of Aggf1 gene and application thereof

Technical Field

The invention relates to the fields of neovascular diseases and nucleic acid medicines, and more particularly relates to a 3' -UTR sequence of an Aggf1 gene and application thereof.

Background

Hypertensive heart disease is a disease with a high current morbidity and mortality. In the course of the disease, hypertensive heart remodeling caused by cardiac inflammation and fibrosis can lead to left ventricular hypertrophy, systolic and diastolic dysfunction, and even heart failure. However, the molecular mechanism of hypertensive heart disease is not yet clear, resulting in no effective treatment regimen currently available.

MicroRNA (miRNA) -mediated modulation of gene expression is a primary method of inhibiting translation and/or promoting degradation of target protein mRNA. It has been reported that some 3 '-terminal non-coding sequences (3' -UTRs) have cis-element effects, can be recognized and bind to mirnas, and thus regulate the stability of their mRNA and protein expression.

Our earlier studies found that AGGF1 protein could improve survival after myocardial infarction in mice. In addition, in the mouse model of myocardial hypertrophy induced by isoproterenol and myocardial hypertrophy induced by abdominal aortic stenosis with pressure overload, exogenous AGGF1 can inhibit myocardial fibrosis and prevent heart dysfunction and myocardial remodeling. In addition, myeloid leukemia 1 (MCL 1) is an anti-apoptotic Bcl2 family protein, highly expressed in the myocardium, MCL1 plays an important role in myocardial homeostasis and autophagy, and the lack of MCL1 leads to impaired autophagy function, mitochondrial dysfunction and fatal heart failure. In patients with heart failure and hypertension, down-regulation of AGGF1 and MCL1 expression occurs. This phenomenon was also found in AngII-treated cardiomyocytes.

Disclosure of Invention

In this study, we found that the Aggf1 level and the Mcl1 level were higher in the body after the introduction of the full-length Aggf1 containing ORF and its 3'-UTR than the introduction of the Aggf1 OFR frame, and that the introduction of a fragment containing only the 3' -UTR of the Aggf1 gene also increased the Aggf1 level and the Mcl1 level at the time of the cell experiment.

Based on the above studies, the present invention provides a nucleic acid element capable of regulating intracellular gene expression, which is characterized by the sequence shown in SEQ ID NO. 1 or 3 or a derivative sequence thereof.

The derivative sequence refers to a sequence which is derived by taking an important site in the sequence shown in SEQ ID NO. 1 or 3 as a core and can be used as a 3' -UTR. For example, in SEQ ID NO. 1, positions 70-74, 129-133, 681-685, 189-192 and 670-173 modulate Aggf1 and Mcl1 levels by binding to the relevant miRNAs; in SEQ ID NO. 3, positions 1966-1972, 1176-1181 and 1747-1752 regulate Aggf1 and Mcl1 levels by binding to related miRNAs, one or more of these positions are kept unchanged, and other positions can be subjected to appropriate point mutation, deletion, insertion, inversion and the like within the scope of the known technology in the art, so long as the regulation function of the 3' -UTR is not affected.

In a preferred embodiment, the nucleic acid element comprises one or more combinations of the first to eighth positions;

the first locus is the 70 th-74 th locus in SEQ ID NO. 1;

the second locus is the 129 th to 133 th positions in SEQ ID NO. 1;

the third locus is 681-685 in SEQ ID NO. 1;

the fourth site is 189-192 in SEQ ID NO. 1;

the fifth locus is 670 th-173 th site in SEQ ID NO. 1;

the sixth site is 1966-1972 in SEQ ID NO. 3;

the seventh locus is the 1176-1181 position in SEQ ID NO. 3;

the eighth site is 1747-1752 in SEQ ID NO. 3.

The first to fifth sites are interaction sites of 3' -UTR of mouse Aggf1 gene and miR105/101/93, and the sixth to eighth sites are interaction sites of 3' -UTR of human Aggf1 gene and miR105/101/93, and other position changes on 3' -UTR do not influence the gene regulation function, but the changes of the sites influence the regulation function.

The invention also provides application of the nucleic acid element in preparing heart reconstruction therapeutic drugs.

The invention also provides application of the nucleic acid element in regulating gene expression in vitro cells.

The invention also provides a nucleic acid construct comprising an ORF and a 3' -UTR, wherein the 3' -UTR is the nucleic acid element described above, the ORF is in tandem with the 3' -UTR, and the 3' -UTR is located at the 3' -end of the ORF.

In a specific embodiment, the ORF is a nucleic acid sequence encoding an Aggf1 protein, and the 3'-UTR is concatenated at the 3' end of the ORF.

Preferably, the ORF sequence is as shown in SEQ ID NO. 2 or 4.

In a specific embodiment, the nucleic acid construct is a double stranded DNA sequence.

The invention also provides application of the nucleic acid construct in preparing a gene therapy medicament. For example, AAV delivery systems can be used to deliver such nucleic acid constructs into cells, or to foci, to modulate expression of related genes, such as Aggf1 and Mcl1, in related cells or tissues, for therapeutic purposes. The nucleic acid element of the invention can be introduced into damaged heart tissue in series with the open reading frame of the Aggf1 gene, which has better therapeutic effect than just introducing the open reading frame.

Drawings

FIG. 1 is a schematic diagram of the construction of AAV systems using different parts of the Aggf1 gene;

FIG. 2 is a photograph of immunoblots of Mcl1 and Aggf1 in AngII perfused mice treated with AAV;

FIG. 3 is a statistical plot of LVFS levels in AngII perfused mice treated with AAV;

FIG. 4 is a statistical plot of HW/BW values and HW/TL values for AngII perfused mice treated with AAV;

FIG. 5 is a statistical plot of the expression levels of Anp, bnp, β -Mhc in heart tissue of AngII perfused mice treated with AAV;

FIG. 6 is a photograph of a masson stain of heart tissue of an AngII perfused mouse with AAV treatment;

FIG. 7 is a TUNEL staining photograph of heart tissue of an AngII perfused mouse with AAV treatment;

FIG. 8 is a statistical plot of the activity of Caspase3 in heart tissue of mice perfused with AAV-treated AngII;

FIG. 9 is a statistical plot of Bax/Bcl2 gene mRNA levels in new cardiomyocytes of mice treated with AngII after various AAV treatments;

FIG. 10 is a photograph of immunoblots of Mcl1 and Aggf1 after treatment of mouse new cardiomyocytes with AAV-Ag-U and a statistical plot of protein levels obtained therefrom;

FIG. 11 is a photograph of immunoblots of Mcl1 and Aggf1 after treatment of wild-type and Aggf1 gene-deleted mouse new cardiomyocytes with AAV-Ag-U;

FIG. 12 is a statistical plot of mRNA levels of Mcl1 and Aggf1 genes in new cardiomyocytes of mice transfected with different miRNAs;

FIG. 13 is a statistical plot of 7 miRNA levels in heart tissue of AngII perfused mice;

FIG. 14 is a statistical plot of 7 miRNA levels in AngII treated mouse new cardiomyocytes;

FIG. 15 is a photograph of immunoblots and mRNA statistics of Aggf1 and Mcl1 after transfection of mouse new cardiomyocytes with three miRNAs alone or in combination;

FIG. 16 is a schematic diagram showing the sites on the 3' -UTR of the Aggf1 gene affecting miRNA binding.

Detailed Description

The principles and features of the present invention are described below with reference to the drawings, the examples are illustrated for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention.

1. Expression of Mcl1 and Aggf1 in cardiac tissue of patients suffering from cardiac dysfunction and hypertension

We collected heart tissue samples from 6 heart transplant patients with heart remodeling symptoms and 3 healthy volunteers, and examined the expression of Mcl1 and Aggf1 genes by RT-PCR, and the results showed that the expression levels of Mcl1 and Aggf1 genes were significantly down-regulated in heart tissues of patients with heart remodeling symptoms, and similar results were obtained by immunoblotting experiments. It can be seen that the down regulation of the expression levels of Mcl1 and Aggf1 genes has a clear correlation with cardiovascular diseases such as heart failure, hypertension, etc.

2. Mouse model

Male C57BL/6 mice, placed in control environment, were conditioned (22.+ -. 1 ℃) and humidity (55%), and were given a 12:12 hour dark photoperiod without restriction of feeding and drinking. All animal studies were approved by the institutional animal care and use committee of the south-middle university and were conducted in accordance with the guidelines of the national institutes of health.

AngII (1 mg/kg/day, sigma) or saline vehicle (Veh) was infused through a transdermal osmotic micropump (Alzet, cupertino, calif.) for 6 weeks to establish a mouse heart reconstruction model.

3. Effect of over-expressed full Length Aggf1 on mouse heart reconstruction model

We used AAV9 drug delivery system heart reconstruction mice for the study. The full-length Aggf1 gene containing the protein coding sequence in Aggf1 and 3'-UTR, the Aggf1 gene containing only the Aggf1 protein coding sequence (Aggf 1, SEQ ID NO:2 or 4, wherein SEQ ID NO:2 is a mouse-derived sequence, SEQ ID NO:4 is a human-derived sequence), and the 3' -UTR sequence (SEQ ID NO:1 or 3, wherein SEQ ID NO:1 is a mouse-derived sequence, SEQ ID NO:3 is a human-derived sequence) were amplified from cDNA and inserted into AAV9 plasmids, respectively, to obtain AAV-Ag-FL, AAV-Ag, and AAV-Ag-U (FIG. 1).

AAV9 was packaged in HEK293 by three plasmid co-transfection and the resulting AAV9 particles were purified. After one week of micropump implantation, AAV-GFP, AAV-Ag or AAV-Ag-FL was injected intravenously once a week for five weeks.

The expression of Aggf1 and Mcl1 genes in the heart tissue of the mice was examined, and the immunoblotting results are shown in FIG. 2, in AngII-perfused mice, both AAV-Ag-treated mice and AAV-Ag-FL-treated mice showed significantly higher expression levels of Aggf1 than AAV-GFP-treated mice, and AAV-Ag-FL induced up-regulated expression of Mcl1 gene, whereas AAV-Ag did not. mRNA level detection also showed similar effects. Echocardiography showed that both AAV-Ag-FL and AAV-Ag treated mice showed higher LVEF values than AAV-GFP treated mice, and AAV-Ag-FL treated mice showed higher LVEF and LVFS values than AAV-Ag (fig. 3).

Furthermore, in AngII mice, AAV-Ag treated mice had significantly reduced heart to body weight ratio (HW/BW) and heart to shin length ratio (HW/TL), while AAV-Ag-FL treated mice had further reduced HW/BW and HW/TL (fig. 4). Similarly, AAV-Ag-FL was also more potent in reducing the expression of Anp, bnp and β -Mhc mRNA than AAV-Ag (FIG. 5). Whereas AAV-Ag and AAV-Ag-FL have little effect on AngII mice in heart rate and blood pressure.

To confirm the effects of AAV-Ag and AAV-Ag-FL, we performed Marsonian staining on mouse heart sections, and the results show in FIG. 6 that AAV-Ag significantly reduced myocardial fibrosis, while AAV-Ag-FL had a more pronounced effect, completely eliminated myocardial fibrosis. Similarly, AAV-Ag and AAV-Ag-FL also inhibit the expression of genes ColIa2, colIVa1, ctgf, mcp1, tnfα and IL6, which are all elevated in AngII perfused mice. Furthermore, the AAV-Ag-FL is also significantly more potent than AAV-Ag. These results show that AAV-Ag-FL has better cardioprotective effects than AAV-Ag.

The effects of AAV-Ag-FL and AAV-Ag on apoptosis in myocardial tissue were examined by TUNEL staining, and as shown in FIG. 7, AAV-Ag significantly reduced apoptosis due to AngII perfusion, while AAV-Ag-FL completely abrogated apoptosis. Furthermore, in AngII perfused mice administered AAV-Ag-FL, the enzyme Caspase3 activity associated with apoptosis was also reduced to levels of non-AngII perfused mice, significantly lower than AngII perfused mice administered AAV-GFP and AAV-Ag (fig. 8).

These results all show that administration of both AAV-Ag-FL and AAV-Ag can inhibit myocardial necrosis in AngII perfused mice and prevent cardiac dysfunction and cardiac remodeling processes. Furthermore, AAV-Ag-FL exhibits more effective cardioprotective function. It can be seen that the 3' -UTR of the Aggf1 gene may play an important role.

4. Cell experiments to observe the influence of AAV-Ag-FL on myocardial cells

Hearts were harvested from 1-3 day old neonatal mice and immediately digested, centrifuged to discard the supernatant, resuspended in growth medium containing 10% serum, cultured for 24h, fresh growth medium was added and cultured for 48h. Then incubated with AAV for 12h. The new cardiomyocytes after AAV incubation were washed with PBS and starved overnight, then treated with AngII for 24h.

TUNEL staining is carried out on the treated new myocardial cells, the apoptosis rate of the new myocardial cells is improved due to the AngII treatment, the apoptosis induced by AngII is inhibited by the AAV-Ag treatment, the effect of AAV-Ag-FL delta is not obviously different from that of AAV-Ag, and the apoptosis rate of the new myocardial cells is recovered to be normal due to the AAV-Ag-FL. Detection of Bax/Bcl2 gene mRNA expression in new cardiomyocytes, consistent with apoptosis staining, resulted in a significant increase in Bax/Bcl2 gene mRNA levels in AngII-treated cells, with AAV-Ag-FL delta and AAV-Ag down-regulating the AngII-induced increase in Bax/Bcl2 gene mRNA levels, while AAV-Ag-FL restored Bax/Bcl2 gene mRNA to normal levels (fig. 9).

These results demonstrate that AAV-Ag-FL has a greater protective effect than AAV-Ag in the response of new cardiomyocytes to AngII-induced stress.

5. Regulatory function of 3' -UTR of Aggf1 Gene and Mcl1 Gene

We have further studied to show that knocking down the Aggf1 gene results in a decrease in Mcl1 gene expression and knocking down the Mcl1 gene results in a decrease in Aggf1 gene expression, but that the respective overexpression of these two genes does not affect the expression of each other, and we hypothesize that the cause of the effect is probably the 3' -UTR of mRNA and involves the ceRNA regulatory mechanism.

To determine whether the 3' -UTR affected the expression of the relevant gene, we constructed AAV system containing only 3' -UTR of Mcl1 (AAV-Mc-U) and AAV system containing only 3' -UTR of Aggf1 (AAV-Ag-U). Treatment of mouse new cardiomyocytes with AAV-Mc-U resulted in ectopic expression of the 3' -UTR of Mcl1, which showed an increase in both the mRNA and protein levels of both the Mcl1 and Aggf1 genes. Similar effects were also produced in new cardiomyocytes in mice treated with AAV-Ag-U (FIG. 10).

The 3' -UTR of human Mcl1 was inserted into the pLuc plasmid to form the pLuc-Mc-U plasmid, and luciferase activity experiments were performed in HEK293 cells, which revealed that the decrease in luciferase activity of pLuc-Mc-U was caused by the siRNA down-regulation of the Aggf1 gene. Similarly, down-regulation of the Mcl1 gene may result in a decrease in luciferase activity of pLuc-Ag-U.

We prepared Aggf1 gene-deleted HEK293 cells (AG-Cas 9) by CRISPR/Cas9 system and then treated the cells with AAV-AG-U, the results showed that Mcl1 gene in AG-Cas9 cells was still up-expressed (fig. 11).

The above experiments demonstrate that the 3' UTR of Aggf1 independently regulates the Mcl1 gene independently of the Aggf1 gene itself. And in human cells, the 3'-UTR of the human Mcl1 and Aggf1 genes has a regulatory function similar to that of the 3' -UTR of the Mcl1 and Aggf1 genes of mice.

6. Identification of miRNAs that interact with the 3' UTR of the Aggf1 Gene and the Mcl1 Gene

We screened 7 miRNAs that were likely to interact with Aggf1 gene and 3' UTR of Mcl1 gene, including miR-495, miR-101, miR-29, miR-105, miR-217, miR-93 and miR-448, by research analysis. These mirnas were transferred into mouse new cardiomyocytes, respectively, and the results showed that miR-101, miR-105 and miR-93 reduced mRNA levels (fig. 12) and protein levels of Aggf1 gene and Mcl1 gene.

In heart tissue of AngII perfused mice and in new cardiomyocytes of mice treated with AngII, we detected up-regulated expression of miR-101, miR-105 and miR-93 (FIGS. 13 and 14). The independent or combined transfection of miR-101, miR-105 and miR-93 into mouse novel heart cells has the advantage that the Aggf1 gene and Mcl1 gene expression levels are significantly reduced in the independent or combined transfected cells. Also, the Aggf1 gene and Mcl1 gene expression levels were greatly reduced in the co-transfected cells compared to the separate transfection (FIG. 15), and it was seen that miR-101, miR-105 and miR-93 had additive effects on the effects of Aggf1 gene and Mcl1 gene expression levels.

The experiment shows that the 3' -UTR of the Aggf1 gene can not only directly act as cis element of mRNA of the Aggf1 gene to improve the expression of the Aggf1 gene and the Mcl1 gene, but also independently improve the expression of the Aggf1 gene and the Mcl1 gene to protect heart tissues and heart cells.

Further experiments and researches show that the 70 th to 74 th and 129 th to 133 th positions of the 3' -UTR of the mouse Aggf1 gene influence the gene regulation function related to miR-105 as shown in FIG. 16; positions 681-685 influence the gene regulation function related to miR-101; the 189 th to 192 th and 670 th to 173 th positions affect the gene regulation function related to miR-93. The 1966-1972 of the 3' -UTR of the human Aggf1 gene influences the gene regulation function related to miR-105; position 1176-1181 affects the gene regulation function related to miR-101; positions 1747-1752 influence the gene regulation function related to miR-93.

The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.

Sequence listing

<110> university of Nanjing medical science

<120> 3' -UTR sequence of Aggf1 gene and use thereof

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gggcttaagc taggacaatc aggccctaaa ctccaaattt ggataaaata tctctttgca 1380

ttcttcttgg ccacctgcat agtctgacac atacgtatgt acagttagac ttgcaggctg 1440

caggagtgcc ctgcattgtt ttcttttaat tagaaataaa aagtattagt ctaaatgtgg 1500

ttctggtgct ggtgccctgt atatatgtaa caatatagga cccctccaaa taggttttgc 1560

ttctggtgaa tcttggtcat tgggttagta tatgactgtc tataatattt tccatttgta 1620

ttattatatt ggggaaaaca ttttttatca ttttctatta agaaaatgga aaacttaaat 1680

atggtgttaa aaatggaaat atatgtgata tttgcattat taaacccctt tgcagatcta 1740

aagtctcagg gttccactat tagtggttaa agtactgaat aatgtctggg tctatttcca 1800

tctgtaaaat aagaatattt gtctcacgtg tctgtcatga atagcaaatg agaaaatgta 1860

tataaaagca ctttgtaaat tgtaaaagta gtacagatgt gagcttctat ttgtttagaa 1920

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cggcaggtgc gggagatcga gaagctgctg catcacacag aacggctgta ccagaacgca 180

gaaagcaaca accaggagct ccgcacgcag gtggaagaac tcagtaaaat actccaacgt 240

gggagaaatg aagataataa aaagtctgat gtagaagtac aaacagagaa ccatgctcct 300

tggtcaatct cagattattt ttatcagacg tactacaatg acgttagtct tccaaataaa 360

gtgactgaac tgtcagatca acaagatcaa gctatcgaaa cttctatttt gaattctaaa 420

gaccatttac aagtagaaaa tgatgcttac cctggtaccg atagaacaga aaatgttaaa 480

tatagacaag tggaccattt tgcctcaaat tcacaggagc cagcatctgc attagcaaca 540

gaagatacct ccttagaagg ctcatcatta gctgaaagtt tgagagctgc agcagaagcg 600

gctgtatcac agactggatt tagttatgat gaaaatactg gactgtattt tgaccacagc 660

actggtttct attatgattc tgaaaatcaa ctctattatg atccttccac tggaatttat 720

tactattgtg atgtggaaag tggtcgttat cagtttcatt ctcgagtaga tttgcaacct 780

tatccgactt ctagcacaaa acaaagtaaa gataaaaaat tgaagaagaa aagaaaagat 840

ccagattctt ctgcaacaaa tgaggaaaag gatttgaact cagaggatca aaaagccttc 900

agtgttgaac atacaagctg caatgaggaa gaaaatttcg caaatatgaa aaagaaggcc 960

aaaataggca ttcatcacaa aaatagtccc cccaaagtca ctgttccaac tagtggaaat 1020

actatagagt ctcctcttca tgaaaacatc tctaattcaa catcatttaa agatgagaaa 1080

atcatggaga ctgatagtga accagaggaa ggtgaaatta cagactctca gactgaggat 1140

agttatgacg aagccattac cagtgaaggc aatgtaactg cagaagatag tgaggatgaa 1200

gatgaagaca aaatttggcc cccatgtatt agagtaattg tcattagatc acctgtgttg 1260

cagataggat cactctttat cattactgct gtaaaccctg ctacaattgg aagagaaaag 1320

gatatggaac atactctccg aatccctgaa gttggtgtca gtaagtttca tgcagaaatt 1380

tattttgacc atgacttaca aagttatgtc cttgtggatc aaggcagtca aaatggcaca 1440

attgttaatg gaaaacagat tcttcagccg aaaactaaat gtgaccctta cgtacttgag 1500

catggagatg aagtcaaaat tggagaaact gtcttatcct ttcacattca tcctggcagt 1560

gatacctgtg atggctgtga accagggcag gttagagccc accttcgcct tgataagaaa 1620

gatgaatctt ttgttggtcc aacactaagt aaggaggaaa aagagttgga aagaagaaaa 1680

gaattaaaga aaatacgagt aaaatatggt ttacagaata cagaatacga agatgaaaag 1740

acattgaaga atccaaaata taaagataga gctggaaaac gtagggagca ggttggaagt 1800

gaaggaactt tccaaagaga tgatgctcct gcatctgttc attctgaaat tactgatagc 1860

aacaaaggtc ggaagatgtt ggagaagatg ggttggaaga aaggagaggg cctggggaag 1920

gatggtggag gaatgaaaac gccgatccag cttcagcttc ggcgaacaca tgcaggcttg 1980

gggacaggca aaccatcctc atttgaagat gttcaccttc tccaaaacaa gaacaaaaaa 2040

aactgggaca aagcacgaga gcggtttact gaaaacttcc cagaaactaa gcctcaaaaa 2100

gatgacccag ggaccatgcc ttgggtaaaa gggactttag agtga 2145

Claims (6)

1. Non-therapeutic use of a nucleic acid element having a sequence as set forth in SEQ ID No. 1 or 3 for regulating the expression of an Aggf1 gene and/or Mcl1 gene in an ex vivo cell.

2. The application of a nucleic acid element in preparing a heart reconstruction therapeutic drug is provided, wherein the sequence of the nucleic acid element is shown as SEQ ID NO. 1 or 3.

3. Non-therapeutic use of a nucleic acid construct comprising an ORF and a 3' -UTR, said 3' -UTR having a sequence as shown in SEQ ID No. 1 or 3, said ORF being concatenated with said 3' -UTR and said 3' -UTR being located at the 3' end of said ORF, said ORF being the nucleic acid sequence encoding an Aggf1 protein, for modulating expression of an Aggf1 gene and/or Mcl1 gene in an ex vivo cell.

4. The use according to claim 3, wherein the ORF sequence is as shown in SEQ ID NO. 2 or 4.

5. The use according to claim 3 or 4, wherein the nucleic acid construct is a double stranded DNA sequence.

6. Use of a nucleic acid construct comprising an ORF and a 3' -UTR, said 3' -UTR having a sequence as shown in SEQ ID No. 1 or 3, said ORF being concatenated with said 3' -UTR and said 3' -UTR being located at the 3' end of said ORF, said ORF being the nucleic acid sequence encoding an Aggf1 protein, in the manufacture of a gene therapeutic for cardiac remodeling.

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