- Email: [email protected]
Cardiomyocytes Derived from Human Embryonic Stem Cells have a Distinctive Micro-RNA Signature
Abstracts CSANZ 2012 Abstracts
BMI between the three cohorts: STEMI 27.45 (95% CI: 26.98–27.93) vs NSTEMI 28.73 (95% CI: 28.52–28.95) vs UA 29.26 kg/m2 (95% CI: 28.84–29.684) p < 0.001. For STEMI patients, 38.07% and 25.87% were overweight and obese respectively, NSTEMI 37.21% and 34.68%, and UA 36.79% and 39.03%, with the lowest proportion of obesity in STEMI patients and highest in UA patients (p < 0.001). Conclusion: Over 70% of ACS patients were either overweight or obese with the highest proportion of obesity amongst those with UA. STEMI patients had the lowest proportion of obesity and the lowest mean BMI. http://dx.doi.org/10.1016/j.hlc.2012.05.648 Cardiovascular Genetics 638 Biomarkers of Thoracic Aneurysm in Heritable Aortopathy Y. Lu ∗ , B. Hambly, R. Jeremy University of Sydney, Australia Tissue studies show that matrix metalloproteinases (MMPs) are activated in thoracic aortic aneurysm, but utility of peripheral blood MMP levels as markers of aneurysm development is not established. Peripheral venous blood levels of MMP2, MMP3 and MMP9 were measured in duplicate (Luminex) in 16 unrelated controls, 38 Marfan (Ghent criteria), 11 nonaneurysmal fibrillinopathy (MASS phenotype) patients and 12 patients with isolated familial aneurysm (FTAD). Variation in repeat MMP measurements was less than 10%. Aortic geometry was measured by echocardiography. Plasma MMP2 levels were increased in Marfan and familial aneurysm patients but not in MASS phenotype. In contrast, plasma MMP3 levels were reduced in Marfan and familial aneurysm patients but were normal in the MASS phenotype group. Controls Age (years) MMP2 (ng/ml) MMP3 (ng/ml) MMP9 (ng/ml) MMP2/MMP3
35 307 13 62 32
± ± ± ± ±
13 51 2 62 27
Marfan 38 449 6 66 92
± ± ± ± ±
14 25* 1** 36 69**
MASS Phenotype 30 322 10 113 47
± ± ± ± ±
19 42 2 65 36
44 473 7 66 66
± ± ± ± ±
19 24* 1* 33 27**
p < 0.025 vs Controls (ANOVA SPSS v 19).
∗∗
p < 0.01 vs Controls (ANOVA SPSS v 19).
In patients with severe aortic dilatation (maximum diameter > 45 mm) plasma MMP2 (518 ± 159 ng/ml) was greater than in those with milder aortic dilatation (405 ± 112 ng/ml, p = 0.02). Plasma MMP2 and MMP3 levels are significantly altered in heritable aortopathy and appear to correlate with severity of aortic disease. http://dx.doi.org/10.1016/j.hlc.2012.05.649
639 Cardiomyocytes Derived from Human Embryonic Stem Cells have a Distinctive Micro-RNA Signature D. Arasaratnam 1,∗ , D. Elliott 1 , K. Koutsis 1 , A. White 1,2 1 Monash 2 Monash
Immunology & Stem Cell Laboratories, Australia Heart Southern Health, Australia
Background and Aim: MicroRNAs (miRs) are small non-coding RNA molecules, which modulate gene expression. A given miR can cause large scale of switching of gene expression programmes, hence may have a role in directing cell fate decisions. Our aim was to identify differentially expressed miRs in human embryonic stem cell (hESC) derived cardiomyocytes. Methods: We had previously generated a reporter cardiac cell line NKX2-5GFP/w in which green fluorescent protein (GFP) expression is under control of the cardiac-specific transcription factor NKX2.5 promoter. This facilitates the separation and comparison of hESC derived cardiomyocytes. NKX2-5GFP/w hESCs were differentiated for seven days, then separated into GFP+ and GFP− fractions by flow cytometry. The differential microRNA expression profile of GFP+ and GFP− fraction was examined by micro-array (Affymetrix) and changes confirmed using RT-PCR Taqman microRNA assays (Applied Biosystems). Results: A number of miRs, previously associated with pluripotency exhibited lower levels in the GFP positive fraction including miRs -200c, -302a, -302c*, -302d, 371-5p, -372, and -373. Similarly, there were a number of miRs with higher expression in the GFP+ fraction, most notably miR-133a, a known muscle marker. The miR-133a result was confirmed by RT-PCR which demonstrated a 70-fold upregulation. Conclusions: Cardiomyocytes derived from human embryonic stem cells have a distinctive miRNA profile. Further functional experiments are required to determine if the differentially expressed miRs are involved in the mechanism of cardiac differentiation.
Isolated FTAD
Mean ± SD. ∗
S265
http://dx.doi.org/10.1016/j.hlc.2012.05.650 640 Characterisation of Pro-Angiogenic Mediators in AgeRelated Impairment Following Ischaemic Injury Y. Lam 1,∗ , L. Lecce 1 , M. Ng 1,2 1 The
Heart Research Institute, Australia Prince Alfred Hospital, Australia
2 Royal
Background: Aging is associated with increased incidence of ischaemic vascular disease and ischaemic mediated angiogenesis is known to impair as a function of age. However, the molecular mechanism(s) of agerelated impairment of angiogenesis are remained to be determined. In this study we aim to characterise factors that contributed to the age-related impaired response by examining the levels of several pro-angiogenic mediators following ischaemic injury in young and aged mice.
ABSTRACTS
Heart, Lung and Circulation 2012;21:S143–S316
BMI between the three cohorts: STEMI 27.45 (95% CI: 26.98–27.93) vs NSTEMI 28.73 (95% CI: 28.52–28.95) vs UA 29.26 kg/m2 (95% CI: 28.84–29.684) p < 0.001. For STEMI patients, 38.07% and 25.87% were overweight and obese respectively, NSTEMI 37.21% and 34.68%, and UA 36.79% and 39.03%, with the lowest proportion of obesity in STEMI patients and highest in UA patients (p < 0.001). Conclusion: Over 70% of ACS patients were either overweight or obese with the highest proportion of obesity amongst those with UA. STEMI patients had the lowest proportion of obesity and the lowest mean BMI. http://dx.doi.org/10.1016/j.hlc.2012.05.648 Cardiovascular Genetics 638 Biomarkers of Thoracic Aneurysm in Heritable Aortopathy Y. Lu ∗ , B. Hambly, R. Jeremy University of Sydney, Australia Tissue studies show that matrix metalloproteinases (MMPs) are activated in thoracic aortic aneurysm, but utility of peripheral blood MMP levels as markers of aneurysm development is not established. Peripheral venous blood levels of MMP2, MMP3 and MMP9 were measured in duplicate (Luminex) in 16 unrelated controls, 38 Marfan (Ghent criteria), 11 nonaneurysmal fibrillinopathy (MASS phenotype) patients and 12 patients with isolated familial aneurysm (FTAD). Variation in repeat MMP measurements was less than 10%. Aortic geometry was measured by echocardiography. Plasma MMP2 levels were increased in Marfan and familial aneurysm patients but not in MASS phenotype. In contrast, plasma MMP3 levels were reduced in Marfan and familial aneurysm patients but were normal in the MASS phenotype group. Controls Age (years) MMP2 (ng/ml) MMP3 (ng/ml) MMP9 (ng/ml) MMP2/MMP3
35 307 13 62 32
± ± ± ± ±
13 51 2 62 27
Marfan 38 449 6 66 92
± ± ± ± ±
14 25* 1** 36 69**
MASS Phenotype 30 322 10 113 47
± ± ± ± ±
19 42 2 65 36
44 473 7 66 66
± ± ± ± ±
19 24* 1* 33 27**
p < 0.025 vs Controls (ANOVA SPSS v 19).
∗∗
p < 0.01 vs Controls (ANOVA SPSS v 19).
In patients with severe aortic dilatation (maximum diameter > 45 mm) plasma MMP2 (518 ± 159 ng/ml) was greater than in those with milder aortic dilatation (405 ± 112 ng/ml, p = 0.02). Plasma MMP2 and MMP3 levels are significantly altered in heritable aortopathy and appear to correlate with severity of aortic disease. http://dx.doi.org/10.1016/j.hlc.2012.05.649
639 Cardiomyocytes Derived from Human Embryonic Stem Cells have a Distinctive Micro-RNA Signature D. Arasaratnam 1,∗ , D. Elliott 1 , K. Koutsis 1 , A. White 1,2 1 Monash 2 Monash
Immunology & Stem Cell Laboratories, Australia Heart Southern Health, Australia
Background and Aim: MicroRNAs (miRs) are small non-coding RNA molecules, which modulate gene expression. A given miR can cause large scale of switching of gene expression programmes, hence may have a role in directing cell fate decisions. Our aim was to identify differentially expressed miRs in human embryonic stem cell (hESC) derived cardiomyocytes. Methods: We had previously generated a reporter cardiac cell line NKX2-5GFP/w in which green fluorescent protein (GFP) expression is under control of the cardiac-specific transcription factor NKX2.5 promoter. This facilitates the separation and comparison of hESC derived cardiomyocytes. NKX2-5GFP/w hESCs were differentiated for seven days, then separated into GFP+ and GFP− fractions by flow cytometry. The differential microRNA expression profile of GFP+ and GFP− fraction was examined by micro-array (Affymetrix) and changes confirmed using RT-PCR Taqman microRNA assays (Applied Biosystems). Results: A number of miRs, previously associated with pluripotency exhibited lower levels in the GFP positive fraction including miRs -200c, -302a, -302c*, -302d, 371-5p, -372, and -373. Similarly, there were a number of miRs with higher expression in the GFP+ fraction, most notably miR-133a, a known muscle marker. The miR-133a result was confirmed by RT-PCR which demonstrated a 70-fold upregulation. Conclusions: Cardiomyocytes derived from human embryonic stem cells have a distinctive miRNA profile. Further functional experiments are required to determine if the differentially expressed miRs are involved in the mechanism of cardiac differentiation.
Isolated FTAD
Mean ± SD. ∗
S265
http://dx.doi.org/10.1016/j.hlc.2012.05.650 640 Characterisation of Pro-Angiogenic Mediators in AgeRelated Impairment Following Ischaemic Injury Y. Lam 1,∗ , L. Lecce 1 , M. Ng 1,2 1 The
Heart Research Institute, Australia Prince Alfred Hospital, Australia
2 Royal
Background: Aging is associated with increased incidence of ischaemic vascular disease and ischaemic mediated angiogenesis is known to impair as a function of age. However, the molecular mechanism(s) of agerelated impairment of angiogenesis are remained to be determined. In this study we aim to characterise factors that contributed to the age-related impaired response by examining the levels of several pro-angiogenic mediators following ischaemic injury in young and aged mice.
ABSTRACTS
Heart, Lung and Circulation 2012;21:S143–S316