Simultaneous Detection and Quantification of Mitochondrial DNA Deletion(s), Depletion, and Over-Replication in Patients with Mitochondrial Disease

Bai, Renkui; Wong, Lee‐Jun C.

doi:10.1016/s1525-1578(10)60595-8

Cited by 145 publications

(120 citation statements)

References 30 publications

Supporting

Mentioning

115

Contrasting

Order By: Relevance

“…The associated increased mtDNA replication observed is known to occur in patients with mtDNA deletions (Bai and Wong 2005). Although increased oxidative stress or oxidative damage was not measured in the tissue, mtDNA overexpression may occur through the reactive oxygen species (ROS)-mediated induction of mtDNA transcription and replication factors (Miranda et al 1999;Lee and Wei 2005).…”

Section: Discussionmentioning

confidence: 99%

Aberrant synthesis of ATP synthase resulting from a novel deletion in mitochondrial DNA in an African patient with progressive external ophthalmoplegia

Westhuizen

Smet

Levanets

et al. 2010

J of Inher Metab Disea

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Aberrant synthesis of ATP synthase resulting from a novel deletion in mitochondrial DNA in an African patient with progressive external ophthalmoplegia

Westhuizen

Smet

Levanets

et al. 2010

J of Inher Metab Disea

View full text Add to dashboard Cite

“…These primer pairs were chosen for regions for which polymorphic variants have not been reported. The MT-TL1 gene region is deleted in Ͻ1.6% of described deletions, and our previous studies have demonstrated that these 2 loci produce the most consistent and reliable results (23 ). Further details of qPCR validation are available in the online Data Supplement.…”

Section: Real-time Qpcr Analysismentioning

confidence: 93%

“…The fluorescence signal intensities of PCR products were recorded and analyzed with a 7900HT Fast Real-Time PCR System (Applied Biosystems) and the manufacturer's Sequence Detection System software (version 2.2.2). mtDNA content was measured by real-time qPCR with primers specific for the regions of mitochondrial gene MT-TL1 [mitochondrially encoded tRNA leucine 1 (UUA/G)] and nuclear gene B2M (beta-2-microglobulin), as previously described (23 ). These primer pairs were chosen for regions for which polymorphic variants have not been reported.…”

Section: Real-time Qpcr Analysismentioning

confidence: 99%

Quantitative Evaluation of the Mitochondrial DNA Depletion Syndrome

et al. 2010

View full text Add to dashboard Cite

Background: The mitochondrial DNA (mtDNA) depletion syndromes (MDDSs) are autosomal recessive disorders characterized by a reduction in cellular mtDNA content. Mutations in at least 9 genes [POLG, polymerase (DNA directed), gamma; DGUOK, deoxyguanosine kinase; TK2, thymidine kinase, mitochondrial; TYMP, thymidine phosphorylase; MPV17, MpV17 mitochondrial inner membrane protein; SUCLA2, succinate-CoA ligase, ADP-forming, beta subunit; SUCLG1, succinate-CoA ligase, alpha subunit; RRM2B, RRM2B, ribonucleotide reductase M2 B (TP53 inducible); and C10orf2, chromosome 10 open reading frame 2 (also known as TWINKLE)] have been reported to cause mtDNA depletion. In the clinical setting, a simple method to quantify mtDNA depletion would be useful before undertaking gene sequence analysis. Methods: Real-time quantitative PCR (qPCR) was used to measure the mtDNA content in blood, muscle, and liver samples and in skin fibroblast cultures from individuals suspected of mitochondrial disorders, with or without deleterious mutations in genes responsible for MDDS. Results: The mtDNA content was quantified in 776 tissue samples (blood, n = 341; muscle, n = 325; liver, n = 63; skin fibroblasts, n = 47) from control individuals. mtDNA content increased with age in muscle tissue, decreased with age in blood samples, and appeared to be unaffected by age in liver samples. In 165 samples (blood, n = 122; muscle, n = 21; liver, n = 15; skin fibroblasts, n = 7) from patients with molecularly proven MDDSs, severe mtDNA depletion was detected in liver and muscle tissue with high specificity and sensitivity. Blood samples were specific but not sensitive for detecting mtDNA depletion, and skin fibroblasts were not valuable for evaluating mtDNA depletion. Mutations in the POLG, RRM2B, and MPV17 genes were prospectively identified in 1 blood, 1 liver, and 3 muscle samples. Conclusions: Muscle and liver tissues, but not blood or skin fibroblasts, are potentially useful for rapid screening for mtDNA depletion with real-time qPCR.

show abstract

“…A fragment of 85 bp from 3' untranslated region of β 2 -microglobulin gene (β 2 -M) was amplified with the forward primer; 5'-TGCT-GTCTCCATGTTTGATGTATCT-3' and the reverse primer; 5'-TCTCTGCTCCCCACCTCTAAGT-3' [32][33][34]. These PCR products were cloned into the pCR 2.1-TOPO vector (Invitrogen).…”

Section: Preparation Of Standard Dna For Quantitative Pcrmentioning

confidence: 99%

“…Each measurement was performed in triplicate and the threshold cycle numbers (C T ) were measured. The copy number was generated from the C T value and standard curve according to previously described procedures [32][33][34].…”

Section: Real-time Quantitative Pcr (Rt Q-pcr)mentioning

confidence: 99%

Endocrine Therapy of Breast Cancer

Clarke¹

2007

View full text Add to dashboard Cite

Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Department of Defense, Washington Headquarters Services, Directorate for Information Operations and Reports (0704-0188), 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302. Respondents should be aware that notwithstanding any other provision of law, no person shall be subject to any penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. REPORT DATE 01-06-20072. REPORT TYPE Annual DATES COVERED PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION REPORT NUMBERGeorgetown University Washington, DC 20007 SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSOR/MONITOR'S ACRONYM(S) U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland 21702-5012 SPONSOR/MONITOR'S REPORT NUMBER(S) DISTRIBUTION / AVAILABILITY STATEMENTApproved for Public Release; Distribution Unlimited SUPPLEMENTARY NOTESOriginal contains colored plates: ALL DTIC reproductions will be in black and white. ABSTRACTA recent controversy in the treatment of estrogen receptor positive (ER+) breast cancers is whether an aromatase inhibitor, e.g., letrozole (LET) or TAM should be given as first line endocrine therapy. Unfortunately, response rates are lower, and response durations are shorter, on crossover than when these agents are given as first line therapies, e.g., ~40% of tumors show crossresistance to TAM or an aromatase inhibitor on crossover. Only 50% of ER+ tumors respond to endocrine therapy. Currently, we fail to predict endocrine responsiveness in about 66% of ER+/PgR-(progesterone receptor), 55% of ER-/PgR+, and 25% of ER+/PgR+ tumors. In this new Clinical Translational Research Award, we hypothesize that our analytical methods can extract expression profiles of breast tumors that define their responsiveness (sensitive vs. resistant) to endocrine therapy. These profiles, when combined with known predictive/prognostic factors, will support neural network and biostatistical classifiers or committee machines that predict each tumor's endocrine responsiveness. Our objectives are to array breast cancer cases, build classifiers of endocrine responsiveness (using microarray data), and validate these classifiers in independent data sets. In the long term, we will design custom arrays for use in clinical practice. Genes will be further studied using cellular and molecular methods, and their role as therapeutic targets explored. SUBJECT TERMSAntiestrogen, aromatase inhibitor, anastrazole, bioinformatics, biomarkers, biostatistics, breast c...

show abstract

Simultaneous Detection and Quantification of Mitochondrial DNA Deletion(s), Depletion, and Over-Replication in Patients with Mitochondrial Disease

Cited by 145 publications

References 30 publications

Aberrant synthesis of ATP synthase resulting from a novel deletion in mitochondrial DNA in an African patient with progressive external ophthalmoplegia

Aberrant synthesis of ATP synthase resulting from a novel deletion in mitochondrial DNA in an African patient with progressive external ophthalmoplegia

Quantitative Evaluation of the Mitochondrial DNA Depletion Syndrome

Endocrine Therapy of Breast Cancer

Contact Info

Product

Resources

About