Teodoro Bottiglieri Ph.D.

Posted September 15th 2019

Maternal glutamine supplementation in murine succinic semialdehyde dehydrogenase deficiency, a disorder of gamma-aminobutyric acid metabolism.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Brown, M. N., D. C. Walters, M. A. Schmidt, J. Hill, A. McConnell, E. E. W. Jansen, G. S. Salomons, E. Arning, T. Bottiglieri, K. M. Gibson and J. B. Roullet (2019). “Maternal glutamine supplementation in murine succinic semialdehyde dehydrogenase deficiency, a disorder of gamma-aminobutyric acid metabolism.” J Inherit Metab Dis 42(5): 1030-1039.

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Murine succinic semialdehyde dehydrogenase deficiency (SSADHD) manifests with high concentrations of gamma-aminobutyric acid (GABA) and gamma-hydroxybutyrate (GHB) and low glutamine in the brain. To understand the pathogenic contribution of central glutamine deficiency, we exposed aldh5a1(-/-) (SSADHD) mice and their genetic controls (aldh5a1(+/+) ) to either a 4% (w/w) glutamine-containing diet or a glutamine-free diet from conception until postnatal day 30. Endpoints included brain, liver and blood amino acids, brain GHB, ataxia scores, and open field testing. Glutamine supplementation did not improve aldh5a1(-/-) brain glutamine deficiency nor brain GABA and GHB. It decreased brain glutamate but did not change the ratio of excitatory (glutamate) to inhibitory (GABA) neurotransmitters. In contrast, glutamine supplementation significantly increased brain arginine (30% for aldh5a1(+/+) and 18% for aldh5a1(-/-) mice), and leucine (12% and 18%). Glutamine deficiency was confirmed in the liver. The test diet increased hepatic glutamate in both genotypes, decreased glutamine in aldh5a1(+/+) but not in aldh5a1(-/-) , but had no effect on GABA. Dried bloodspot analyses showed significantly elevated GABA in mutants (approximately 800% above controls) and decreased glutamate (approximately 25%), but no glutamine difference with controls. Glutamine supplementation did not impact blood GABA but significantly increased glutamine and glutamate in both genotypes indicating systemic exposure to dietary glutamine. Ataxia and pronounced hyperactivity were observed in aldh5a1(-/-) mice but remained unchanged by the diet intervention. The study suggests that glutamine supplementation improves peripheral but not central glutamine deficiency in experimental SSADHD. Future studies are needed to fully understand the pathogenic role of brain glutamine deficiency in SSADHD.


Posted August 15th 2019

Gamma-Hydroxybutyrate content in dried bloodspots facilitates newborn detection of succinic semialdehyde dehydrogenase deficiency.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Brown, M., P. Ashcraft, E. Arning, T. Bottiglieri, J. B. Roullet and K. M. Gibson (2019). “Gamma-Hydroxybutyrate content in dried bloodspots facilitates newborn detection of succinic semialdehyde dehydrogenase deficiency.” Mol Genet Metab Jul 18. [Epub ahead of print].

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Increased gamma-hydroxybutyric acid in urine and blood are metabolic hallmarks of succinic semialdehyde dehydrogenase deficiency, a defect of 4-aminobutyric acid metabolism. Here, we examined the hypothesis that succinic semialdehyde dehydrogenase deficiency could be identified via measurement of gamma-hydroxybutyric acid in newborn and post-newborn dried bloodspots. Quantitation of gamma-hydroxybutyric acid using liquid chromatography-tandem mass spectrometry in twelve archival newborn patient dried bloodspots was 360+/-57muM (mean, standard error; range 111-767), all values exceeding the previously established cutoff for newborn detection of 78 muMu established from 2831 dried bloodspots derived from newborns, neonates and children. Gamma-hydroxybutyric acid in post-newborn dried bloodspots (n=19; ages 0.8-38years) was 191+/-65muM (mean, standard error; range 20-1218), exceeding the aforementioned GHB cutoff for patients approximately 10years of age or younger. Further, gamma-hydroxybutyric acid in post-newborn dried bloodspots displayed a significant (p<.0001) inverse correlation with age. This preliminary study suggests that succinic semialdehyde dehydrogenase deficiency may be identified in newborn and post-newborn dried bloodspots via quantitation of gamma-hydroxybutyric acid, while forming the platform for more extensive studies in affected and unaffected dried bloodspots.


Posted August 15th 2019

Betaine attenuates pathology by stimulating lipid oxidation in liver and regulating phospholipid metabolism in brain of methionine-choline-deficient rats.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Abu Ahmad, N., M. Raizman, N. Weizmann, B. Wasek, E. Arning, T. Bottiglieri, O. Tirosh and A. M. Troen (2019). “Betaine attenuates pathology by stimulating lipid oxidation in liver and regulating phospholipid metabolism in brain of methionine-choline-deficient rats.” FASEB 33(8): 9334-9349.

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Methyl-donor deficiency is a risk factor for neurodegenerative diseases. Dietary deficiency of the methyl-donors methionine and choline [methionine-choline-deficient (MCD) diet] is a well-established model of nonalcoholic steatohepatitis (NASH), yet brain metabolism has not been studied in this model. We hypothesized that supplemental betaine would protect both the liver and brain in this model and that any benefit to the brain would be due to improved liver metabolism because betaine is a methyl-donor in liver methylation but is not metabolically active in the brain. We fed male Sprague-Dawley rats a control diet, MCD diet, or betaine-supplemented MCD (MCD+B) diet for 8 wk and collected blood and tissue. As expected, betaine prevented MCD diet-induced NASH. However, contrary to our prediction, it did not appear to do so by stimulating methylation; the MCD+B diet worsened hyperhomocysteinemia and depressed liver methylation potential 8-fold compared with the MCD diet. Instead, it significantly increased the expression of genes involved in beta-oxidation: fibroblast growth factor 21 and peroxisome proliferator-activated receptor alpha. In contrast to that of the liver, brain methylation potential was unaffected by diet. Nevertheless, several phospholipid (PL) subclasses involved in stabilizing brain membranes were decreased by the MCD diet, and these improved modestly with betaine. The protective effect of betaine is likely due to the stimulation of beta-oxidation in liver and the effects on PL metabolism in brain.-Abu Ahmad, N., Raizman, M., Weizmann, N., Wasek, B., Arning, E., Bottiglieri, T., Tirosh, O., Troen, A. M. Betaine attenuates pathology by stimulating lipid oxidation in liver and regulating phospholipid metabolism in brain of methionine-choline-deficient rats.


Posted July 15th 2019

5-Methyltetrahydrofolate in Maternal Diets Alters DNA Methylation Potential and Increases Later Life Weight Gain and Food Intake in Wistar Rat Dams and Female Offspring (P11-022-19).

Erland Arning Ph.D.

Erland Arning Ph.D.

Pannia, E., R. Hammoud, R. Kubant, J. Sa, N. Yang, M. Ho, D. Chatterjee, Z. Pausova, E. Arning, T. Bottiglieri and G. H. Anderson (2019). “5-Methyltetrahydrofolate in Maternal Diets Alters DNA Methylation Potential and Increases Later Life Weight Gain and Food Intake in Wistar Rat Dams and Female Offspring (P11-022-19).” Curr Dev Nutr Volume 3, Issue Supplement 1, June 2019. [Epub ahead of print].

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Objectives: Diet during pregnancy programs the mother and offspring post-weaning (PW). Folic acid (FA, synthetic folate) mediates DNA methylation (DNAm) reactions and high intakes, simulating those consumed by American women, lead to epigenetic dysregulation of energy metabolic pathways. 5-methyltetrahydrofolate (5MTHF), the bioactive folate form, has gained popularity as a supplement due to direct cellular uptake and utilization and does not increase unmetabolized FA (UMFA). However, a comparison of folate forms on in utero programming of offspring or maternal health has not been reported. Our objectives were to compare the effects of folate dose (low vs high) and form (FA vs 5MTHF) during pregnancy on DNAm potential, and the early and later PW phenotype of Wistar rat mothers and female offspring (mothers-to-be). Methods: Pregnant Wistar rats (n = 22/group) were fed an AIN93G diet with recommended FA (1X, 2mg/kg diet), 5X-FA or equimolar 5MTHF. Dams were fed 1X-FA during lactation and then dams and female pups were fed a high fat diet for 19 weeks. Weight gain (WG), food intake (FI), energy expenditure (EE), insulin resistance (IR), plasma 5MTHF, UMFA, homocysteine (tHcy), and hepatic S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH) and DNA methyltransferases (DNMT) activity at birth and PW were measured. Results: Dams fed 5MTHF diets had lower DNMT activity at birth and female pups had higher SAM: SAH ratios (P < 0.05) indicative of altered DNAm potential compared to FA diets. Plasma 5MTHF at birth was dose dependent with 5X diets leading to higher levels than 1X diets (P < 0.001) in dams and pups. In contrast, UMFA was only higher in 5X-FA dams. 5MTHF dams had higher tHcy at birth and were more IR at 1 week PW than FA fed dams (P < 0.05). In both dams and offspring, high 5MTHF also led to higher WG (> 15%, P < 0.001) and FI (> 5%, P < 0.001) compared to high FA diets up to 19 weeks. EE (P < 0.05) was higher suggesting a compensatory response to WG. 5X-MTHF dams, but not offspring, also had greater hepatic lipids (P < 0.05) than other groups. Conclusions: Folate dose and form during pregnancy affects DNAm potential at birth and early and later phenotype of dams and female offspring. High 5MTHF increases WG, FI and hepatic lipids PW suggesting it may not be the preferred form for prenatal supplements or additions to the food supply. Funding Sources: Supported by CIHR-INMD; EP supported by NSERC-CGS D.


Posted July 15th 2019

Choline and Folic Acid Balance in Diets During Pregnancy Programs Food Intake Regulation in Wistar Rat Offspring (FS08-05-19).

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Hammoud, R., C. S. Liao, E. Pannia, M. Ho, N. Yang, R. Kubant, D. Chatterjee, E. Arning, T. Bottiglieri, Z. Pausova and G. H. Anderson (2019). “Choline and Folic Acid Balance in Diets During Pregnancy Programs Food Intake Regulation in Wistar Rat Offspring (FS08-05-19).” Curr Dev Nutr Jun 13: 3(Suppl 1). [Epub ahead of print].

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Objectives: High gestational folic acid (FA) induces an obesogenic phenotype in male Wistar rat offspring. Imbalances between FA and other methyl-nutrients (i.e., choline) leading to perturbations in the 1-carbon cycle may account for the effects of high FA diets. Canadian women consume high (2-7-fold) intakes of FA, but most are not meeting recommended adequate intakes for choline. Choline is also absent from Canadian prenatal supplements. The objective of this study is to evaluate the effects of the interaction between choline and FA in maternal diets of rats on the 1-carbon cycle, and the programming of food intake, body weight gain and biomarkers of obesity in the offspring later in life. Methods: Pregnant Wistar rat dams were fed the AIN-93 G diet with recommended (1X) choline and FA (RCRF, control), or a 5X FA diet with either 0.5X choline (LCHF), 1X choline (RCHF), or 2.5X choline (HCHF). Brain and blood were collected at birth. At weaning one male pup/dam from all groups was maintained on the control diet for 20 weeks then terminated. Dependent measures include weekly body weight-gain and food intake, plasma glucoregulatory hormones and 1-carbon metabolites at birth and post-weaning. Results: Increasing choline content to 2.5-fold in a high (5-fold) gestational FA diet (HCHF) led to lower plasma insulin and leptin levels at birth compared to the LCHF and RCHF diets, respectively (P < 0.05). It also led to lower (25%, P = 0.03) plasma 5-methyltetrahydrofolate concentrations at birth compared to the RCHF diet, suggesting more efficient utilization of FA. Offspring born to dams maintained on a high folic acid diet with either low or recommended choline had higher weekly food intake (6%, P < 0.05) and body weight-gain (9%, P < 0.01). In contrast, offspring from dams fed the HCHF gestational diet were not different from those born to dams fed the RCRF (control) diet, highlighting the mitigating effects of a balanced choline and FA gestational diet. Conclusions: Increased intakes of choline mitigate the effects of high FA diets. Maternal dietary choline interacts with FA on the long-term programming of food intake regulation in the offspring; emphasizing a need for more attention to improving choline intakes by women of child-bearing age. Funding Sources: This research was funded by the Canadian Institute of Health Research, Institute of Nutrition, Metabolism and Diabetes (CIHR-INMD).