Proliferation kinetics of subpopulations of human marrow cells determined by quantifying in vivo incorporation of [2H2]-glucose into DNA of S-phase cells

Schwartz, Gretchen N.; Vance, Barbara A.; Levine, Benjamin; Fukazawa, Motoharu; Telford, William G.; Cesar, Denise; Hellerstein, Marc K.; Gress, Ronald E.

doi:10.1182/blood-2003-01-0139

Cited by 21 publications

(17 citation statements)

References 25 publications

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“…Our results show that CD133+ cells were located predominantly in layer III of the human SVZ, and within the central region of layer I-III in the RMS. Although the function of CD133 is uncertain, it is often considered to be a multipotent 'stem cell' marker, and is expressed in carcinogenic stem cells (Pilkington, 2005;Song et al, 2008), human embryonic stem cells (Barraud et al, 2007;Uchida et al, 2000) and haematopoetic progenitor cells (Schwartz et al, 2003;Wagner et al, 2004). The expression of CD133+ cells throughout the RMS, together with its co-expression with PCNA, supports the notion that these cells may be stem or progenitor cells distributed throughout the extent of the human RMS.…”

Section: Discussionmentioning

confidence: 78%

The cellular composition and morphological organization of the rostral migratory stream in the adult human brain

Kam

Curtis

McGlashan

et al. 2009

Journal of Chemical Neuroanatomy

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Section: Discussionmentioning

confidence: 78%

The cellular composition and morphological organization of the rostral migratory stream in the adult human brain

Kam

Curtis

McGlashan

et al. 2009

Journal of Chemical Neuroanatomy

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“…Sim- plete loss of function in the PU.1/CEBPA/RUNX1 circuit, since experimentally, complete inactivation of PU.1, CEBPA, or RUNX1 kills AML cells, even as partial loss of function of any one of these is leukemogenic (17,43,(68)(69)(70)(71)(72); and accordingly, NPM1, RUNX1, and biallelic CEBPA mutations, though highly recurrent in AML, are mutually exclusive (37)(38)(39). PU.1, CEBPA, and RUNX1 have been shown to promote exponential replication kinetics by binding to MYC enhancers to produce high-grade activation of MYC (the master transcription factor coordinator of cell proliferation) and by co-binding with MYC at its target genes (73)(74)(75)(76)(77)(78)(79)(80)(81)(82). This contrasts with the quiescence imposed by stem cell master transcription factors such as HLF in HSCs (73)(74)(75)(76)(77)(78)(79)(80)(81)(82).…”

Section: Resistance In Vivo and In Vitro Was By Avoidance Of Pharmacomentioning

confidence: 99%

“…PU.1, CEBPA, and RUNX1 have been shown to promote exponential replication kinetics by binding to MYC enhancers to produce high-grade activation of MYC (the master transcription factor coordinator of cell proliferation) and by co-binding with MYC at its target genes (73)(74)(75)(76)(77)(78)(79)(80)(81)(82). This contrasts with the quiescence imposed by stem cell master transcription factors such as HLF in HSCs (73)(74)(75)(76)(77)(78)(79)(80)(81)(82). In computational analyses, such skewing of intrinsic replication rates logarithmically favors decoupling replication from forward differentiation in lineage progenitors as the most efficient strategy for malignant transformation (83).…”

Section: Resistance In Vivo and In Vitro Was By Avoidance Of Pharmacomentioning

confidence: 99%

Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates

Gu¹,

Ebrahem²,

Mahfouz³

et al. 2018

Journal of Clinical Investigation

115

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“…To measure the turnover of DNA [1,2] or cells [3][4][5] in various tissues, DNA can be labeled with isotope-labeled precursor. In such studies, the DNA is extracted, hydrolyzed, and analyzed to determine label incorporation by mass spectrometry.…”

Section: Introductionmentioning

confidence: 99%

DNA digestion to deoxyribonucleoside: A simplified one-step procedure

Quinlivan

Gregory

2008

Analytical Biochemistry

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We present a simple and inexpensive 'one-step' protocol for the hydrolysis of DNA to deoxyribonucleosides. Unlike the older DNA hydrolysis protocol which is cumbersome and labor intensive, thes new protocol is ideal for high-throughput assays and is suitable automation. Using this protocol we were able to hydrolyze several hundred samples within an 8-hour period. The new protocol is fully compatible with LC-MS/MS and gives similar recoveries for all five major deoxyribonucleosides when compared to the older protocol.The requirement to hydrolyze DNA to deoxyribonucleosides is a common component of several assays. To measure the turnover of DNA [1,2] or cells [3][4][5] in various tissues, DNA can be labeled with isotope-labeled precursor. In such studies, the DNA is extracted, hydrolyzed, and analyzed to determine label incorporation by mass spectrometry. Likewise, in vivo experiments in humans, such as examination of the effect of gene polymorphisms [6] and nutritional status [6,7] on DNA metabolism, have used labeled one-carbon donors to label monocyte DNA. Nucleoside hydrolysates of DNA also have been used in determining the base composition of DNA, including investigation of epigenetic modification such as deoxycytosine methylation [8,9], oxidative damage [10,11], or unique DNA composition [12].One of the most commonly used protocols for digesting DNA is the tri-enzyme protocol devised by Crain [13]. However, a major weakness of the Crain protocol is that the first enzyme in the reaction, nuclease P1 (E.C. 3.1.30.1), only can digest single stranded DNA and has a pH optimum (pH ~5) significantly lower and a reaction temperature significantly higher (~50°C) than the other two enzymes (pH >8 and ~37°C) in the reaction. This leads to an overly complex protocol [Fig 1], whereby the DNA must be boiled to denature (and rapidly cooled to prevent reversion), the pH of the sample is adjusted twice, and the sample undergoes three separate enzyme incubations. These complexities limit the number of samples that can be prepared. For logistical reasons (repetitive manipulations of tubes), we were typically [6] restricted to preparing ~60 samples per day when we used the Crain protocol. Furthermore, the Crain protocol uses high buffer concentrations (e.g., 50-100 mmol/L ammonium bicarbonate, pH 7.9) which can interfere with downstream reactions and assays. Therefore, we devised a simple *Corresponding Author: Email: epq@ufl.edu (E.P. Quinlivan). 1 Abbreviations Used: LC-MS/MS: liquid chromatography -tandem mass spectrometry; H-ESI: heated electrospray ionization; dCyt: deoxycytidine; MdCyt: 5-methyldeoxycytidine; dThy: thymidine; dAdn: deoxyadenosine; dGua: deoxyguanosine Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that dur...

show abstract

Proliferation kinetics of subpopulations of human marrow cells determined by quantifying in vivo incorporation of [2H2]-glucose into DNA of S-phase cells

Cited by 21 publications

References 25 publications

The cellular composition and morphological organization of the rostral migratory stream in the adult human brain

The cellular composition and morphological organization of the rostral migratory stream in the adult human brain

Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates

DNA digestion to deoxyribonucleoside: A simplified one-step procedure

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