Teodoro Bottiglieri Ph.D.

Posted September 16th 2021

Metabolomic Profiling of Adults with Congenital Heart Disease.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Cedars, A., C. Manlhiot, J. M. Ko, T. Bottiglieri, E. Arning, A. Weingarten, A. Opotowsky and S. Kutty (2021). “Metabolomic Profiling of Adults with Congenital Heart Disease.” Metabolites 11(8).

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Metabolomic analysis may provide an integrated assessment in genetically and pathologically heterogeneous populations. We used metabolomic analysis to gain mechanistic insight into the small and diverse population of adults with congenital heart disease (ACHD). Consecutive ACHD patients seen at a single institution were enrolled. Clinical variables and whole blood were collected at regular clinical visits. Stored plasma samples were analyzed for the concentrations of 674 metabolites and metabolic markers using mass spectrometry with internal standards. These samples were compared to 28 simultaneously assessed healthy non-ACHD controls. Principal component analysis and multivariable regression modeling were used to identify metabolites associated with clinical outcomes in ACHD. Plasma from ACHD and healthy control patients differed in the concentrations of multiple metabolites. Differences between control and ACHD were greater in number and in degree than those between ACHD anatomic groups. A metabolite cluster containing amino acids and metabolites of amino acids correlated with negative clinical outcomes across all anatomic groups. Metabolites in the arginine metabolic pathway, betaine, dehydroepiandrosterone, cystine, 1-methylhistidine, serotonin and bile acids were associated with specific clinical outcomes. Metabolic markers of disease may both be useful as biomarkers for disease activity and suggest etiologically related pathways as possible targets for disease-modifying intervention.


Posted July 15th 2021

Pharmacodynamics of cerebrospinal fluid asparagine after asparaginase.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Panetta, J.C., Liu, Y., Bottiglieri, T., Arning, E., Cheng, C., Karol, S.E., Yang, J.J., Zhou, Y., Inaba, H., Pui, C.H., Jeha, S. and Relling, M.V. (2021). “Pharmacodynamics of cerebrospinal fluid asparagine after asparaginase.” Cancer Chemother Pharmacol Jun 25. [Epub ahead of print].

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PURPOSE: We evaluated effects of asparaginase dosage, schedule, and formulation on CSF asparagine in children with acute lymphoblastic leukemia (ALL). METHODS: We evaluated CSF asparagine (2114 samples) and serum asparaginase (5007 samples) in 482 children with ALL treated on the Total XVI study (NCT00549848). Patients received one or two 3000 IU/m(2) IV pegaspargase doses during induction and were then randomized in continuation to receive 2500 IU/m(2) or 3500 IU/m(2) IV intermittently (four doses) on the low-risk (LR) or continuously (15 doses) on the standard/high risk (SHR) arms. A pharmacokinetic-pharmacodynamic model was used to estimate the duration of CSF asparagine depletion below 1 uM. RESULTS: During induction, CSF asparagine depletion after two doses of pegaspargase was twice as long as one dose (median 30.7 vs 15.3 days, p < 0.001). During continuation, the higher dose increased the CSF asparagine depletion duration by only 9% on the LR and 1% in the SHR arm, consistent with the nonlinear pharmacokinetics of serum asparaginase. Pegaspargase caused a longer CSF asparagine depletion duration (1.3-5.3-fold) compared to those who were switched to erwinase (p < 0.001). The median (quartile range) serum asparaginase activity needed to maintain CSF asparagine below 1 µM was 0.44 (0.20, 0.99) IU/mL. Although rare, CNS relapse was higher with decreased CSF asparagine depletion (p = 0.0486); there was no association with relapse at any site (p = 0.3). CONCLUSIONS: The number of pegaspargase doses has a stronger influence on CSF asparagine depletion than did dosage, pegaspargase depleted CSF asparagine longer than erwinase, and CSF asparagine depletion may prevent CNS relapses.


Posted June 17th 2021

Moderate Folic Acid Supplementation in Pregnant Mice Results in Altered Methyl Metabolism and in Sex-specific Placental Transcription Changes.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Luan, Y., Leclerc, D., Cosín-Tomás, M., Malysheva, O.V., Wasek, B., Bottiglieri, T., Caudill, M.A. and Rozen, R. (2021). “Moderate Folic Acid Supplementation in Pregnant Mice Results in Altered Methyl Metabolism and in Sex-specific Placental Transcription Changes.” Mol Nutr Food Res May 19;e2100197. [Epub ahead of print]. e2100197.

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SCOPE: Many pregnant women have higher folic acid (FA) intake due to food fortification and increased vitamin use. We reported that diets containing 5-fold higher FA than recommended for mice (5xFASD) during pregnancy, resulted in methylenetetrahydrofolate reductase (MTHFR) deficiency and altered choline/methyl metabolism, with neurobehavioral abnormalities in newborns. Our goal was to determine whether these changes had their origins in the placenta during embryonic development. METHODS AND RESULTS: Female mice were fed control diet (CD) or 5xFASD for a month before mating and maintained on these diets until embryonic day 17.5. 5xFASD led to pseudo-MTHFR deficiency in maternal liver and altered choline/methyl metabolites in maternal plasma (increased methyltetrahydrofolate and decreased betaine). Methylation potential (S-adenosylmethionine: S-adenosylhomocysteine ratio) and glycerophosphocholine were decreased in placenta and embryonic liver. FASD resulted in sex-specific transcriptome profiles in placenta, with validation of dietary expression changes of 29 genes involved in angiogenesis, receptor biology or neurodevelopment, and altered methylation of the serotonin receptor 2A gene. CONCLUSION: Moderate increases in folate intake during pregnancy result in placental metabolic and gene expression changes, particularly in angiogenesis, which may contribute to abnormal behavior in pups. These results are relevant for determining a safe upper limit for folate intake during pregnancy.


Posted June 17th 2021

Analysis of differential neonatal lethality in cystathionine β-synthase deficient mouse models using metabolic profiling.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Gupta, S., Wang, L., Slifker, M.J., Cai, K.Q., Maclean, K.N., Wasek, B., Bottiglieri, T. and Kruger, W.D. (2021). “Analysis of differential neonatal lethality in cystathionine β-synthase deficient mouse models using metabolic profiling.” Faseb j 35(6): e21629.

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Cystathionine beta-synthase (CBS) is a key enzyme of the trans-sulfuration pathway that converts homocysteine to cystathionine. Loss of CBS activity due to mutation results in CBS deficiency, an inborn error of metabolism characterized by extreme elevation of plasma total homocysteine (tHcy). C57BL6 mice containing either a homozygous null mutation in the cystathionine β-synthase (Cbs(-/-) ) gene or an inactive human CBS protein (Tg-G307S Cbs(-/-) ) are born in mendelian numbers, but the vast majority die between 18 and 21 days of age due to liver failure. However, adult Cbs null mice that express a hypomorphic allele of human CBS as a transgene (Tg-I278T Cbs(-/-) ) show almost no neonatal lethality despite having serum tHcy levels similar to mice with no CBS activity. Here, we characterize liver and serum metabolites in neonatal Cbs(+/-) , Tg-G307S Cbs(-/-) , and Tg-I278T Cbs(-/-) mice at 6, 10, and 17 days of age to understand this difference. In serum, we observe similar elevations in tHcy in both Tg-G307S Cbs(-/-) and Tg-I278T Cbs(-/-) compared to control animals, but methionine is much more severely elevated in Tg-G307S Cbs(-/-) mice. Large scale metabolomic analysis of liver tissue confirms that both methionine and methionine-sulfoxide are significantly more elevated in Tg-G307S Cbs(-/-) animals, along with significant differences in several other metabolites including hexoses, amino acids, other amines, lipids, and carboxylic acids. Our data are consistent with a model that the neonatal lethality observed in CBS-null mice is driven by excess methionine resulting in increased stress on a variety of related pathways including the urea cycle, TCA cycle, gluconeogenesis, and phosphatidylcholine biosynthesis.


Posted May 21st 2021

High Intakes of [6S]-5-Methyltetrahydrofolic Acid Compared with Folic Acid during Pregnancy Programs Central and Peripheral Mechanisms Favouring Increased Food Intake and Body Weight of Mature Female Offspring.

Teodoro Bottiglieri, Ph.D.

Teodoro Bottiglieri, Ph.D.

Pannia, E., Hammoud, R., Kubant, R., Sa, J.Y., Simonian, R., Wasek, B., Ashcraft, P., Bottiglieri, T., Pausova, Z. and Anderson, G.H. (2021). “High Intakes of [6S]-5-Methyltetrahydrofolic Acid Compared with Folic Acid during Pregnancy Programs Central and Peripheral Mechanisms Favouring Increased Food Intake and Body Weight of Mature Female Offspring.” Nutrients 13(5).

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Supplementation with [6S]-5-methyltetrahydrofolic acid (MTHF) is recommended as an alternative to folic acid (FA) in prenatal supplements. This study compared equimolar gestational FA and MTHF diets on energy regulation of female offspring. Wistar rats were fed an AIN-93G diet with recommended (2 mg/kg diet) or 5-fold (5X) intakes of MTHF or FA. At weaning, female offspring were fed a 45% fat diet until 19 weeks. The 5X-MTHF offspring had higher body weight (>15%), food intake (8%), light-cycle energy expenditure, and lower activity compared to 5X-FA offspring (p < 0.05). Both the 5X offspring had higher plasma levels of the anorectic hormone leptin at birth (60%) and at 19 weeks (40%), and lower liver weight and total liver lipids compared to the 1X offspring (p < 0.05). Hypothalamic mRNA expression of leptin receptor (ObRb) was lower, and of suppressor of cytokine signaling-3 (Socs3) was higher in the 5X-MTHF offspring (p < 0.05), suggesting central leptin dysregulation. In contrast, the 5X-FA offspring had higher expression of genes encoding for dopamine and GABA- neurotransmitter receptors (p < 0.01), consistent with their phenotype and reduced food intake. When fed folate diets at the requirement level, no differences were found due to form in the offspring. We conclude that MTHF compared to FA consumed at high levels in the gestational diets program central and peripheral mechanisms to favour increased weight gain in the offspring. These pre-clinical findings caution against high gestational intakes of folates of either form and encourage clinical trials examining their long-term health effects when consumed during pregnancy.