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MS529 FOLATE-PEG-SUPERPARAMAGNETIC IRON OXIDE NANOPARTICLES LABELED WITH CY5.5 FOR LUNG CANCER IMAGING
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Atherosclerosis Supplements 11, no. 2 (2010) 109–222
MS528 INCREASED PLASMA LEVELS OF NON-HDL-BOUND SPHINGOSINE-1-PHOSPHATE (S1P) IN CORONARY ARTERY DISEASE ARE CAUSED BY ALTERATIONS OF S1P UPTAKE BY HDL K. Sattler1 , S. Elbasan1 , P. Keul1 , M. Elter-Schulz2 , C. Bode3 , M. Graler ¨ 3, 4 M. Brocker-Preuss ¨ , T. Budde5 , R. Erbel6 , G. Heusch1 , B. Levkau1 . 1 Institute of Pathophysiology, University Hospital Essen, 2 Institute for Transfusion Medicine, University Duisburg-Essen, Essen, 3 Institute for Immunology, Hannover Medical School, Hannover, 4 Department of Endocrinology, Division of Clinical Chemistry and Laboratory, University Duisburg-Essen, 5 Clinic of Internal Medicine and Cardiology, Alfried Krupp Hospital, 6 Clinic of Cardiology, West German Heart Center, University Duisburg-Essen, Essen, Germany Objective: High-density lipoproteins (HDL) are the major plasma carriers for S1P in healthy individuals but their S1P content is unknown for patients with coronary artery disease (CAD). Methods: S1P was determined in plasma and in HDL isolated from patients with myocardial infarction (MI, n = 83), stable CAD (sCAD, n = 95) and controls (n = 85). The S1P-uptake capacity of HDL isolated from CAD patients and controls as well as of modified isolated HDL and of HDL subfractions was assessed in vitro. Results: HDL-bound plasma S1P levels were dependent on the plasma HDL-C levels. HDL-bound plasma S1P was 1.5-fold lower and non-HDL-bound S1P 8-fold higher in MI and sCAD compared to controls. Non-HDL-bound plasma S1P correlated with CAD symptom severity (CCS) and discriminated patients from controls in ROC analyses. The S1P content of isolated HDL was inversely associated with the non-HDL-bound plasma S1P only in controls but not in CAD patients. In vitro, HDL from controls but not from CAD patients acquired exogenous S1P proportionally to their initial S1P content. The uptake of S1P by isolated HDL was dramatically reduced by chlorination or oxidation, and was 3.8-fold less for HDL3 than HDL2. Conclusions: HDL-bound S1P is decreased and non-HDL-bound S1P increased in CAD. Lower plasma HDL-C levels, altered HDL subfractions and CAD-related modifications of HDL such as chlorination and oxidation may affect the S1P uptake by HDL in CAD. MS529 FOLATE-PEG-SUPERPARAMAGNETIC IRON OXIDE NANOPARTICLES LABELED WITH CY5.5 FOR LUNG CANCER IMAGING M. Islam1 , M.-K. Yoo1 , H.-T. Lim2 , H.-L. Jiang2 , S.-J. Lee3 , I.-K. Park3 , M.-H. Cho2 , C.-S. Cho1 . 1 Department of Agricultural Biotechnology, 2 College of Veterinary Medicine, Seoul National University, Seoul, 3 Department of Biomedical Sciences, Chonnam National University Medical School, Gwangju, Republic of Korea Aims: We have developed a functionalized iron oxide nanoprobe for lung cancer imaging. The nanoprobe was fabricated by immobilizing the folate-conjugated poly(ethylene glycol) (Fol-PEG) onto the surface of superparamagnetic iron oxide nanoparticles (SPIONs) and by labeling with near-infrared (NIR) fluorescence dye Cy5.5. Here, PEG was introduced to prevent the aggregation of SPIONs in physiological medium and capturing of these species by the cells of reticuloendothelial system (RES) and to increase circulation time in the body. Folate conjugation was aimed at tumor-specific nanoprobe targeting through the folate-receptor-mediated endocytosis because folate receptor is often elevated in several cancers including lung cancer. Also, Cy5.5 was used as a fluorophore for optical imaging. Methods: The particle size, morphology, and crystallinity of Fol-PEG-SPIONs were characterized by electrophoretic light scattering spectrophotometer, field emission scanning electron microscopy, and X-ray diffractometer, respectively. In vitro intracellular uptake of the Fol-PEG-SPIONs-Cy5.5 was assessed by FACS. Their toxicity was also assessed in vitro and in vivo. Further, we checked in vivo imaging after their intravenous administration in lung cancer model mice. After systemic injection of Fol-PEG-SPIONs-Cy5.5, bio-distribution profiles in various tissues were quantitatively determined and visualized. Results: Fol-PEG-SPIONs-Cy5.5 exhibited excellent stability in physiological medium, lower cytoxicity, and enhanced targeted delivery efficiency to cancer cells both in vitro and in vivo. NIR fluorescence imaging demonstrated that tumor sites were better clearly identified upon systemic injection of Fol-PEGSPIONs-Cy5.5 than PEG-SPIONs-Cy5.5 without folate. Conclusion: The above results suggest that Fol-PEG-SPIONs-Cy5.5 can be exploited as in vivo optical imaging agent for lung cancer diagnosis.
Memory Stick Presentations
MS530 NEW LPL AND APOA5 GEN MUTATIONS IDENTIFIED IN SUBJECTS WITH SEVERE HYPERTRIGLYCERIDEMIA 3 ´ I. De Castro Oros ´ 1 , A.L. Garc´ıa Ot´ın2 , M. Pueyo1 , P. Mozas1 , L. Gomez ´ , 5 ´ F. Fabiani4 , L. Alvarez-Sala , E. Ros6 , D. Zambon ´ 6 , J. Dalmau7 , F. Civeira2 , 8 1 M. Pocov´ı . RECAVA, Dpto. Bioqu´ımica y Biolog´ıa Molecular y Celular, ´ de Ciencias de la Salud Universidad de Zaragoza. Instituto Aragones ´ Molecular, Hospital (I+CS), 2 Unidad de L´ıpidos. Laboratorio de Investigacion ´ de Ciencias de la Salud (I+CS), Universitario Miguel Servet. Insituto Aragones ´ de Gastroenterolog´ıa, Hepatolog´ıa y Nutricion, ´ Hospital Zaragoza, 3 Seccion San Juan de dios, Barcelona, 4 UGC de Bioqu´ımica Cl´ınica, Hospital Virgen de ˜ la Macarena, Sevilla, 5 Unidad de L´ıpidos, Hospital Gregorio Maranon, Madrid, 6 ´ Institut d’Investigacions Unitat de Lipids, Servei d’Endocrinologia i Nutricio., ` Biomediques August Pi i Sunyer (IDIBAPS), Hospital Cl´ınic and Ciber ´ (CEBEROBN), Barcelona, 7 Unidad Fisiopatolog´ıa de la Obesidad y Nutricion ´ y Metabolopat´ıas, Hospital Infantil de la Fe, Valencia, 8 CIBERER, de Nutricion Dpto. Bioqu´ımica y Biolog´ıa Molecular y Celular, Universidad de Zaragoza. ´ de Ciencias de la Salud (I+CS), Zaragoza, Spain Instituto Aragones Introduction: Several genes have been associated with severe Hypertriglyceriemia (SHTG) Lipoprotein lipase (LpL) is a critical enzyme in triglyceride (TG) metabolism. Apolipoprotein C-II (Apo C-II) is required by LpL for its activation. Apo-AV is also needed to regulate TG metabolism. Nowadays, more than 100 mutations have been described in LPL, APOC2 and APOA5 associated with SHTG. Objective: The aim of our study was to identify the genetic causes of SHTG. Methods: We selected 33 unrelated with TG levels higher than 900 mg/dl and without secondary causes of SHTG. Complete sequencing of exons and intronexon boundaries of LPL, APOC2 and APOA5 was carried out. Results: We found 5 variants in LPL gene not previously described: H80R (c.907>G), Q262X (c.1452C>T), K287_A288del (c.1502_1507delAAGGCC), G300V (c.1542G>T) and R333H (c.1666G>A); and one in APOA5, L253P. No mutations in APOC2 were found. Moreover, 3 patients presented mutations in LPL previously described, two G215E homozygous and one I232S homozygous. The Q262X mutation was found in homozygosis, whereas G300V and R333H variants were found in heterozygosis. The individual who presented the K287_A288del mutation was also carrier of a synonymous variant: T388T. Furthermore, the L253P in APOA5 was found in homozygosis. Analyzing these mutations in a normolipidemic control group (100 alleles) revealed no carriers of these mutations. Conclusions: We have identified SHTG causes in 8 out of 33 (24%) subjects: 5 new and 2 recurrent mutations in LPL and one mutation in APOA5. MS531 POOR MENTAL HEALTH, HPA AXIS REACTIVITY AND SUBCLINICAL CORONARY CALCIFICATION A. Seldenrijk1,2 , M. Hamer2 , B. Penninx1,3,4 , A. Steptoe2 . 1 Psychiatry, VU University Medical Center, Amsterdam, The Netherlands, 2 Epidemiology and Public Health, University College London, London, UK, 3 Psychiatry, Leiden University Medical Center, Leiden, 4 Psychiatry, University Medical Center Groningen, Groningen, The Netherlands Objective: Poor mental health has been associated with coronary heart disease (CHD). One of the hypothesized mechanisms is HPA-axis dysfunction. We examined the associations between poor mental health, cortisol response to laboratory-induced mental stress and subclinical coronary artery calcification (CAC). Methods: Participants were 527 CHD-free volunteers (mean age = 63.0±5.7 years), drawn from the Whitehall II cohort. CAC was measured using electron beam computed tomography. Mental health was assessed using the Short Form-36 Mental Health subscale at time of the heart scan (current) and additionally five times over preceding 15 years (chronic). Salivary cortisol was measured in response to mental stressors (Stroop, mirror tracing). Results: Detectable CAC (some: Agatston score 1–399; severe: Agatston 400) was found in 56.4% of the sample. After adjustment for sociodemographic and conventional risk factors, chronic but not current poor mental health was associated with a higher risk of severe CAC (per SD decrease: ORsevere = 1.47, 95% CI = 1.02–2.12; log Agatston = 0.25±0.11, p = 0.02 within detectable CAC subpopulation). Poor mental health was associated with blunted cortisol response (per SD decrease: bcurrent = −0.06, p = 0.01; bchronic = −0.05, p = 0.03). We observed a trend for interaction (p = 0.097) indicating that persons with poor mental health and high cortisol reactivity showed the highest severe CAC prevalence. Conclusions: In healthy, older participants, chronic but not current poor mental health is associated with CAC. Although participants with poor mental health generally showed blunted cortisol responses, those with cortisol responsiveness were at higher risk for severe CAC.
Atherosclerosis Supplements 11, no. 2 (2010) 109–222
MS528 INCREASED PLASMA LEVELS OF NON-HDL-BOUND SPHINGOSINE-1-PHOSPHATE (S1P) IN CORONARY ARTERY DISEASE ARE CAUSED BY ALTERATIONS OF S1P UPTAKE BY HDL K. Sattler1 , S. Elbasan1 , P. Keul1 , M. Elter-Schulz2 , C. Bode3 , M. Graler ¨ 3, 4 M. Brocker-Preuss ¨ , T. Budde5 , R. Erbel6 , G. Heusch1 , B. Levkau1 . 1 Institute of Pathophysiology, University Hospital Essen, 2 Institute for Transfusion Medicine, University Duisburg-Essen, Essen, 3 Institute for Immunology, Hannover Medical School, Hannover, 4 Department of Endocrinology, Division of Clinical Chemistry and Laboratory, University Duisburg-Essen, 5 Clinic of Internal Medicine and Cardiology, Alfried Krupp Hospital, 6 Clinic of Cardiology, West German Heart Center, University Duisburg-Essen, Essen, Germany Objective: High-density lipoproteins (HDL) are the major plasma carriers for S1P in healthy individuals but their S1P content is unknown for patients with coronary artery disease (CAD). Methods: S1P was determined in plasma and in HDL isolated from patients with myocardial infarction (MI, n = 83), stable CAD (sCAD, n = 95) and controls (n = 85). The S1P-uptake capacity of HDL isolated from CAD patients and controls as well as of modified isolated HDL and of HDL subfractions was assessed in vitro. Results: HDL-bound plasma S1P levels were dependent on the plasma HDL-C levels. HDL-bound plasma S1P was 1.5-fold lower and non-HDL-bound S1P 8-fold higher in MI and sCAD compared to controls. Non-HDL-bound plasma S1P correlated with CAD symptom severity (CCS) and discriminated patients from controls in ROC analyses. The S1P content of isolated HDL was inversely associated with the non-HDL-bound plasma S1P only in controls but not in CAD patients. In vitro, HDL from controls but not from CAD patients acquired exogenous S1P proportionally to their initial S1P content. The uptake of S1P by isolated HDL was dramatically reduced by chlorination or oxidation, and was 3.8-fold less for HDL3 than HDL2. Conclusions: HDL-bound S1P is decreased and non-HDL-bound S1P increased in CAD. Lower plasma HDL-C levels, altered HDL subfractions and CAD-related modifications of HDL such as chlorination and oxidation may affect the S1P uptake by HDL in CAD. MS529 FOLATE-PEG-SUPERPARAMAGNETIC IRON OXIDE NANOPARTICLES LABELED WITH CY5.5 FOR LUNG CANCER IMAGING M. Islam1 , M.-K. Yoo1 , H.-T. Lim2 , H.-L. Jiang2 , S.-J. Lee3 , I.-K. Park3 , M.-H. Cho2 , C.-S. Cho1 . 1 Department of Agricultural Biotechnology, 2 College of Veterinary Medicine, Seoul National University, Seoul, 3 Department of Biomedical Sciences, Chonnam National University Medical School, Gwangju, Republic of Korea Aims: We have developed a functionalized iron oxide nanoprobe for lung cancer imaging. The nanoprobe was fabricated by immobilizing the folate-conjugated poly(ethylene glycol) (Fol-PEG) onto the surface of superparamagnetic iron oxide nanoparticles (SPIONs) and by labeling with near-infrared (NIR) fluorescence dye Cy5.5. Here, PEG was introduced to prevent the aggregation of SPIONs in physiological medium and capturing of these species by the cells of reticuloendothelial system (RES) and to increase circulation time in the body. Folate conjugation was aimed at tumor-specific nanoprobe targeting through the folate-receptor-mediated endocytosis because folate receptor is often elevated in several cancers including lung cancer. Also, Cy5.5 was used as a fluorophore for optical imaging. Methods: The particle size, morphology, and crystallinity of Fol-PEG-SPIONs were characterized by electrophoretic light scattering spectrophotometer, field emission scanning electron microscopy, and X-ray diffractometer, respectively. In vitro intracellular uptake of the Fol-PEG-SPIONs-Cy5.5 was assessed by FACS. Their toxicity was also assessed in vitro and in vivo. Further, we checked in vivo imaging after their intravenous administration in lung cancer model mice. After systemic injection of Fol-PEG-SPIONs-Cy5.5, bio-distribution profiles in various tissues were quantitatively determined and visualized. Results: Fol-PEG-SPIONs-Cy5.5 exhibited excellent stability in physiological medium, lower cytoxicity, and enhanced targeted delivery efficiency to cancer cells both in vitro and in vivo. NIR fluorescence imaging demonstrated that tumor sites were better clearly identified upon systemic injection of Fol-PEGSPIONs-Cy5.5 than PEG-SPIONs-Cy5.5 without folate. Conclusion: The above results suggest that Fol-PEG-SPIONs-Cy5.5 can be exploited as in vivo optical imaging agent for lung cancer diagnosis.
Memory Stick Presentations
MS530 NEW LPL AND APOA5 GEN MUTATIONS IDENTIFIED IN SUBJECTS WITH SEVERE HYPERTRIGLYCERIDEMIA 3 ´ I. De Castro Oros ´ 1 , A.L. Garc´ıa Ot´ın2 , M. Pueyo1 , P. Mozas1 , L. Gomez ´ , 5 ´ F. Fabiani4 , L. Alvarez-Sala , E. Ros6 , D. Zambon ´ 6 , J. Dalmau7 , F. Civeira2 , 8 1 M. Pocov´ı . RECAVA, Dpto. Bioqu´ımica y Biolog´ıa Molecular y Celular, ´ de Ciencias de la Salud Universidad de Zaragoza. Instituto Aragones ´ Molecular, Hospital (I+CS), 2 Unidad de L´ıpidos. Laboratorio de Investigacion ´ de Ciencias de la Salud (I+CS), Universitario Miguel Servet. Insituto Aragones ´ de Gastroenterolog´ıa, Hepatolog´ıa y Nutricion, ´ Hospital Zaragoza, 3 Seccion San Juan de dios, Barcelona, 4 UGC de Bioqu´ımica Cl´ınica, Hospital Virgen de ˜ la Macarena, Sevilla, 5 Unidad de L´ıpidos, Hospital Gregorio Maranon, Madrid, 6 ´ Institut d’Investigacions Unitat de Lipids, Servei d’Endocrinologia i Nutricio., ` Biomediques August Pi i Sunyer (IDIBAPS), Hospital Cl´ınic and Ciber ´ (CEBEROBN), Barcelona, 7 Unidad Fisiopatolog´ıa de la Obesidad y Nutricion ´ y Metabolopat´ıas, Hospital Infantil de la Fe, Valencia, 8 CIBERER, de Nutricion Dpto. Bioqu´ımica y Biolog´ıa Molecular y Celular, Universidad de Zaragoza. ´ de Ciencias de la Salud (I+CS), Zaragoza, Spain Instituto Aragones Introduction: Several genes have been associated with severe Hypertriglyceriemia (SHTG) Lipoprotein lipase (LpL) is a critical enzyme in triglyceride (TG) metabolism. Apolipoprotein C-II (Apo C-II) is required by LpL for its activation. Apo-AV is also needed to regulate TG metabolism. Nowadays, more than 100 mutations have been described in LPL, APOC2 and APOA5 associated with SHTG. Objective: The aim of our study was to identify the genetic causes of SHTG. Methods: We selected 33 unrelated with TG levels higher than 900 mg/dl and without secondary causes of SHTG. Complete sequencing of exons and intronexon boundaries of LPL, APOC2 and APOA5 was carried out. Results: We found 5 variants in LPL gene not previously described: H80R (c.907>G), Q262X (c.1452C>T), K287_A288del (c.1502_1507delAAGGCC), G300V (c.1542G>T) and R333H (c.1666G>A); and one in APOA5, L253P. No mutations in APOC2 were found. Moreover, 3 patients presented mutations in LPL previously described, two G215E homozygous and one I232S homozygous. The Q262X mutation was found in homozygosis, whereas G300V and R333H variants were found in heterozygosis. The individual who presented the K287_A288del mutation was also carrier of a synonymous variant: T388T. Furthermore, the L253P in APOA5 was found in homozygosis. Analyzing these mutations in a normolipidemic control group (100 alleles) revealed no carriers of these mutations. Conclusions: We have identified SHTG causes in 8 out of 33 (24%) subjects: 5 new and 2 recurrent mutations in LPL and one mutation in APOA5. MS531 POOR MENTAL HEALTH, HPA AXIS REACTIVITY AND SUBCLINICAL CORONARY CALCIFICATION A. Seldenrijk1,2 , M. Hamer2 , B. Penninx1,3,4 , A. Steptoe2 . 1 Psychiatry, VU University Medical Center, Amsterdam, The Netherlands, 2 Epidemiology and Public Health, University College London, London, UK, 3 Psychiatry, Leiden University Medical Center, Leiden, 4 Psychiatry, University Medical Center Groningen, Groningen, The Netherlands Objective: Poor mental health has been associated with coronary heart disease (CHD). One of the hypothesized mechanisms is HPA-axis dysfunction. We examined the associations between poor mental health, cortisol response to laboratory-induced mental stress and subclinical coronary artery calcification (CAC). Methods: Participants were 527 CHD-free volunteers (mean age = 63.0±5.7 years), drawn from the Whitehall II cohort. CAC was measured using electron beam computed tomography. Mental health was assessed using the Short Form-36 Mental Health subscale at time of the heart scan (current) and additionally five times over preceding 15 years (chronic). Salivary cortisol was measured in response to mental stressors (Stroop, mirror tracing). Results: Detectable CAC (some: Agatston score 1–399; severe: Agatston 400) was found in 56.4% of the sample. After adjustment for sociodemographic and conventional risk factors, chronic but not current poor mental health was associated with a higher risk of severe CAC (per SD decrease: ORsevere = 1.47, 95% CI = 1.02–2.12; log Agatston = 0.25±0.11, p = 0.02 within detectable CAC subpopulation). Poor mental health was associated with blunted cortisol response (per SD decrease: bcurrent = −0.06, p = 0.01; bchronic = −0.05, p = 0.03). We observed a trend for interaction (p = 0.097) indicating that persons with poor mental health and high cortisol reactivity showed the highest severe CAC prevalence. Conclusions: In healthy, older participants, chronic but not current poor mental health is associated with CAC. Although participants with poor mental health generally showed blunted cortisol responses, those with cortisol responsiveness were at higher risk for severe CAC.