Identification and characterization of potent, selective, and efficacious inhibitors of human arylamine N-acetyltransferase 1

Leggett, Carmine; Doll, Mark A.; Salazar‐González, Raúl A.; Habil, Mariam R.; Trent, John O.; Hein, David W.

doi:10.1007/s00204-021-03194-x

Cited by 16 publications

(15 citation statements)

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“…Small molecule inhibitors, gene-specific siRNAs/shRNAs, and CRISPR/Cas9 knockout (KO) approaches have been used to inhibit or deplete NAT1 in both breast cancer and other cancer cell types. 7,[11][12][13][14] One of the most reproducible findings from these studies is that inhibition or depletion of NAT1 result in a significant decrease in anchorageindependent cell growth in breast cancer cell lines. 7,11,12 This indicates that NAT1 positively contributes to the growth and survival of breast cancer cells and may be essential for the cells to maintain resistance to anoikis, thereby promoting metastasis.…”

Section: Introductionmentioning

confidence: 89%

“…In recent years, we and others have investigated the role of NAT1 in breast cancer using different in vitro model systems. Small molecule inhibitors, gene‐specific siRNAs/shRNAs, and CRISPR/Cas9 knockout (KO) approaches have been used to inhibit or deplete NAT1 in both breast cancer and other cancer cell types 7,11–14 . One of the most reproducible findings from these studies is that inhibition or depletion of NAT1 result in a significant decrease in anchorage‐independent cell growth in breast cancer cell lines 7,11,12 .…”

Section: Introductionmentioning

confidence: 99%

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Deletion of arylamine N‐acetyltransferase 1 in MDA‐MB‐231 human breast cancer cells reduces primary and secondary tumor growth in vivo with no significant effects on metastasis

Doll

Ray

Salazar‐González

et al. 2022

Molecular Carcinogenesis

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Arylamine N‐acetyltransferase 1 (NAT1) is frequently upregulated in breast cancer. Previous studies showed that inhibition or depletion of NAT1 in breast cancer cells diminishes anchorage‐independent growth in culture, suggesting that NAT1 contributes to breast cancer growth and metastasis. To further investigate the contribution of NAT1 to growth and cell invasive/migratory behavior, we subjected parental and NAT1 knockout (KO) breast cancer cell lines (MDA‐MB‐231, MCF‐7, and ZR‐75‐1) to multiple assays. The rate of cell growth in suspension was not consistently decreased in NAT1 KO cells across the cell lines tested. Similarly, cell migration and invasion assays failed to produce reproducible differences between the parental and NAT1 KO cells. To overcome the limitations of in vitro assays, we tested parental and NAT1 KO cells in vivo in a xenograft model by injecting cells into the flank of immunocompromised mice. NAT1 KO MDA‐MB‐231 cells produced primary tumors smaller than those formed by parental cells, which was contributed by an increased rate of apoptosis in KO cells. The frequency of lung metastasis, however, was not altered in NAT1 KO cells. When the primary tumors of the parental and NAT1 KO cells were allowed to grow to a pre‐determined size or delivered directly via tail vein, the number and size of metastatic foci in the lung did not differ between the parental and NAT1 KO cells. In conclusion, NAT1 contributes to primary and secondary tumor growth in vivo in MDA‐MB‐231 breast cancer cells but does not appear to affect its metastatic potential.

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Section: Introductionmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Deletion of arylamine N‐acetyltransferase 1 in MDA‐MB‐231 human breast cancer cells reduces primary and secondary tumor growth in vivo with no significant effects on metastasis

Doll

Ray

Salazar‐González

et al. 2022

Molecular Carcinogenesis

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“…DNA adducts not repaired can result in mutations leading to cancer ( Wang et al, 2019 ; NTP, 2021 ). NAT1 catalyzes ABP N-acetyltransferase and N-OH-ABP O-acetyltransferase activities ( Hein et al, 1993 ; Millner et al, 2012 ; Zhou et al, 2013 ; Leggett et al, 2021 ; Leggett et al, 2022 ; Hein et al, 2022 ).…”

Section: Introductionmentioning

confidence: 99%

“…Many arylamines and alkylanilines undergo N- and O-acetylation catalyzed by NAT1 ( Minchin et al, 1992 ; Hein et al, 1993 ; Zenser et al, 1996 ; Liu et al, 2007 ; Leggett et al, 2021 ). R127 and Y129 amino acids in NAT1 reduce the volume of the NAT1 binding pocket by ∼40% compared to NAT2 ( Wu et al, 2007 ; Zhou et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

560G>A (rs4986782) (R187Q) Single Nucleotide Polymorphism in Arylamine N-Acetyltransferase 1 Increases Affinity for the Aromatic Amine Carcinogens 4-Aminobiphenyl and N-Hydroxy-4-Aminobiphenyl: Implications for Cancer Risk Assessment

Doll

Hein

2022

Front. Pharmacol.

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Human arylamine N-acetyltransferase 1 (NAT1) catalyzes the N-acetylation of arylamine carcinogens such as 4-aminobiphenyl (ABP), and following N-hydroxylation, the O-acetylation of N-hydroxy-arylamine carcinogens such as N-hydroxy-ABP (N-OH-ABP). Genetic polymorphisms in NAT1 are linked to cancer susceptibility following exposures. The effects of individual single nucleotide polymorphisms (SNPs) in the NAT1 coding exon on Michaelis-Menten kinetic constants was assessed for ABP N-acetyltransferase and N-OH-ABP O-acetyltransferase activity following transfection of human NAT1 into COS-1 cells (SV40-transformed African green monkey kidney cells). NAT1 coding region SNPs 97C > T (rs56318881) (R33stop), 190C > T (rs56379106) (R64W), 559C > T (rs5030839) (R187stop) and 752A > T (rs56172717) (D251V) reduced ABP N- acetyltransferase and N-OH-ABP O-acetyltransferase activity below detection. 21T > G (rs4986992) (synonymous), 402T > C (rs146727732) (synonymous), 445G > A (rs4987076) (V149I), 613A > G (rs72554609) (M205V) and 640T > G (rs4986783) (S241A) did not significantly affect Vmax for ABP N-acetyltransferase or N-OH-ABP O-acetyltransferase. 781G > A (rs72554610) (E261K), and 787A > G (rs72554611) (I263V) slightly reduced ABP N-acetyltransferase and N-OH-ABP O-acetyltransferase activities whereas 560G > A (rs4986782) (R187Q) substantially and significantly reduced them. 560G > A (rs4986782) (R187Q) significantly reduced the apparent Km for ABP and N-OH-ABP a finding that was not observed with any of the other NAT1 SNPs tested. These findings suggest that the role of the 560G > A (rs4986782) (R187Q) SNP cancer risk assessment may be modified by exposure level to aromatic amine carcinogens such as ABP.

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“…Aromatic amines undergo hepatic hydroxylation catalyzed by CYP1A2 ( Bartsch et al, 2000 ). O -acetylation of these hydroxylated aromatic amines by NAT1 leads to formation of reactive electrophiles that covalently bind to nucleophilic sites on proteins, DNA and RNA, forming adducts potentially leading to mutagenesis and carcinogenesis ( Barnes et al, 2018 ; Leggett et al, 2022 ).…”

Section: Introductionmentioning

confidence: 99%

Acetyl coenzyme A kinetic studies on N-acetylation of environmental carcinogens by human N-acetyltransferase 1 and its NAT1*14B variant

Habil

Doll²,

Hein³

2022

Front. Pharmacol.

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N-acetyltransferase 1 (NAT1) is a xenobiotic metabolizing enzyme that uses acetyl coenzyme A (AcCoA) as a cofactor for N-acetylation of many carcinogens including aromatic amines and alkylanilines. NAT1 is characterized by single nucleotide polymorphisms (SNPs) that may modulate affinity towards AcCoA. In the current study, we used Chinese hamster ovary (CHO) cells stably transfected with human NAT1*4 (reference allele) or NAT1*14B (variant allele) to measure AcCoA kinetic parameters for N-acetyltransferase activity measurements towards p-aminobenzoic acid (PABA), 4-aminobiphenyl (4-ABP), β-naphthylamine (BNA), benzidine and 3,4-dimethylaniline (3,4-DMA). Our results showed higher N-acetylation rates for each substrate catalyzed by NAT1*4 compared to NAT1*14B. NAT1*4 exhibited higher affinity to AcCoA when catalyzing the N-acetylation of BNA and benzidine compared to NAT1*14B. The results of the current study provide further insights into differences in carcinogen metabolism among individuals possessing the NAT1*14B haplotype.

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Identification and characterization of potent, selective, and efficacious inhibitors of human arylamine N-acetyltransferase 1

Cited by 16 publications

References 50 publications

Deletion of arylamine N‐acetyltransferase 1 in MDA‐MB‐231 human breast cancer cells reduces primary and secondary tumor growth in vivo with no significant effects on metastasis

Deletion of arylamine N‐acetyltransferase 1 in MDA‐MB‐231 human breast cancer cells reduces primary and secondary tumor growth in vivo with no significant effects on metastasis

560G>A (rs4986782) (R187Q) Single Nucleotide Polymorphism in Arylamine N-Acetyltransferase 1 Increases Affinity for the Aromatic Amine Carcinogens 4-Aminobiphenyl and N-Hydroxy-4-Aminobiphenyl: Implications for Cancer Risk Assessment

Acetyl coenzyme A kinetic studies on N-acetylation of environmental carcinogens by human N-acetyltransferase 1 and its NAT1*14B variant

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