Asparagine synthetase as a causal, predictive biomarker for <scp>l</scp>-asparaginase activity in ovarian cancer cells

Lorenzi, Philip L.; Reinhold, William C.; Rudelius, Martina; Gunsior, Michele; Shankavaram, Uma; Bussey, Kimberly J.; Scherf, Uwe; Eichler, Gabriel S.; Martin, Scott E.; Chin, Koei; Gray, Joe W.; Kohn, Elise C.; Horak, Ivan D.; Hoff, Daniel D. Von; Raffeld, Mark; Goldsmith, Paul K.; Caplen, Natasha J.; Weinstein, John N.

doi:10.1158/1535-7163.mct-06-0447

Cited by 94 publications

(76 citation statements)

References 46 publications

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“…What is more, for NK/T cell lymphoma, we did not examine the genetic aberrations, and whether there is a potential relationship between gene fusion or other subtypes of aberrations and ASNS expression. Pancreatic and ovarian tumors and HCC with low ASNS expression levels have also been proposed for L-Asp therapy (17,20,29). In addition, one study revealed that this drug has an antiproliferative effect in β-catenin-mutated HepG2 cells (30).…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, the expression and the functional significance of ASNS have been investigated in solid tumors. ASNS was considered as a causal, predictive biomarker for L-Asp activity in ovarian cancer cells (16,17). Furthermore, one study also showed that the enhanced expression of ASNS protected pancreatic cancer cells from apoptosis induced by glucose deprivation and cisplatin (18).…”

Section: Introductionmentioning

confidence: 99%

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Asparagine synthetase expression and its potential prognostic value in patients with NK/T cell lymphoma

Zhang

et al. 2014

Oncology Reports

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Asparagine synthetase expression and its potential prognostic value in patients with NK/T cell lymphoma

Zhang

et al. 2014

Oncology Reports

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“…Twenty micrograms of total protein per lane was electrophoresed in sodium dodecyl sulfate (SDS)-polyacrylamide, transferred to Immun-Blot polyvinylidene difluoride membrane (Bio-Rad), and probed with antibodies against ASNS or b-actin (Sigma) as we reported previously. 25 Relative ASNS and b-actin expression levels were quantified using ImageJ. …”

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confidence: 99%

The glutaminase activity of l-asparaginase is not required for anticancer activity against ASNS-negative cells

Chan

Lorenzi

Anishkin

et al. 2014

Blood

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• We used molecular dynamics, saturation mutagenesis, and enzymologic screening to develop a glutaminase-free mutant (Q59L) L-ASP.• We then used Q59L to show that glutaminase activity is not required for L-ASP activity against ASNS-negative cancer cells.L-Asparaginase (L-ASP) is a key component of therapy for acute lymphoblastic leukemia. Its mechanism of action, however, is still poorly understood, in part because of its dual asparaginase and glutaminase activities. Here, we show that L-ASP's glutaminase activity is not always required for the enzyme's anticancer effect. We first used molecular dynamics simulations of the clinically standard Escherichia coli L-ASP to predict what mutated forms could be engineered to retain activity against asparagine but not glutamine. Dynamic mapping of enzyme substrate contacts identified Q59 as a promising mutagenesis target for that purpose. Saturation mutagenesis followed by enzymatic screening identified Q59L as a variant that retains asparaginase activity but shows undetectable glutaminase activity. Unlike wild-type L-ASP, Q59L is inactive against cancer cells that express measurable asparagine synthetase (ASNS). Q59L is potently active, however, against ASNS-negative cells. Those observations indicate that the glutaminase activity of L-ASP is necessary for anticancer activity against ASNS-positive cell types but not ASNS-negative cell types.Because the clinical toxicity of L-ASP is thought to stem from its glutaminase activity, these findings suggest the hypothesis that glutaminase-negative variants of L-ASP would provide larger therapeutic indices than wild-type L-ASP for ASNS-negative cancers. (Blood. 2014;123(23):3596-3606) Introduction L-Asparaginase (L-ASP) is an enzyme drug used in combination with vincristine and a glucocorticoid (eg, dexamethasone) to treat acute lymphoblastic leukemia (ALL).1,2 We 3-6 and others 7 have reported a rationale for testing L-ASP against low-asparagine synthetase (ASNS) solid tumors as well. L-ASP's primary known enzymatic activity is deamidation of asparagine to aspartic acid and ammonia, but it also deamidates glutamine to glutamic acid and ammonia, although with lower affinity and lower maximal rate. L-ASP therapy is often limited by toxic side effects that are generally attributed to the glutaminase activity. 8,9 Those side effects often preclude completion of the full treatment regimen, resulting in poor outcome. 10 The question that arises, however, is whether the therapeutic index of L-ASP could be increased by decreasing its glutaminase activity 8,9 or whether that would also decrease the anticancer effect commensurately.One side of the debate hypothesizes that L-ASP's therapeutic index can be improved by increasing glutaminase activity. In support of that hypothesis, data collected over the last decade have suggested that glutaminase activity generally increases the efficacy of L-ASP and is sometimes required to achieve an anticancer effect. Those studies have reported asparaginase activity to be expendable. [11][12][13][...

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“…Lymphoid origin and solid tumors were prone to Lasparginase activity and showed resistant to the drugs among them soft tissue sarcoma [41], ovarian cancer [45], β-catenin mutated hepatocellular carcinoma [42], hepatocellular carcinoma with low expression of asparagine synthetase [43], and gastric adenocarcinoma [44] were studied. From the literature surveyed and reviewed L-asparaginase exhibited potent anti neoplastic and anti lymphomatic activity against tumors [1], [46], [29], [32], [40] This enzyme causes selective death of asparagines dependent tumor cells and also induces apoptosis in tumor cells [26].…”

Section: Assessment Of Anti-proliferative Activitymentioning

confidence: 99%

Purification and Cytotoxicity of L-asparaginase from Streptomyces griseoluteus GDJ1

2017

IJSR

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L-asparaginase (L-asparagine amidohydrolase E.C.3.5.1.1) is an effective antineoplastic enzyme, used in acute lymphoblastic leukemia (ALL) chemotherapy. L-asparaginase activity of marine actinomycetes in soil samples of Agnitheertham (Rameswaram) and Periyar lake, Kumily were investigated. Totally fifty one actinomycetes were isolated from both the soil samples in different selective media. Out of fifty one isolates, four were found to produce L-asparaginase. Among them, the strain AT-R-S1 produced high amount of L-asparaginase. Hence it was selected for the further identification and based on molecular identification the strain AT-R-S1 was identified as Streptomyces griseoluteus GDJ1. Purification of the enzyme from the strain S.griseoluteus GDJ1 (AT-R-S1) was carried out. The final purification of the enzyme in CM-Sephadex C-50 column chromatography yielded 1.27 U/ml of enzyme in 0.09 mg of protein with a specific activity of 14.11 U/mg with approximately 119 folds purity. Cytotoxicity of different concentration of purified L-asparaginase was determined on VERO cells. The anti proliferative activity of L-asparaginase in MCF-7 and HeLa cells was determined and the EC 50 value was found to be 31.37 and 38.99 µg respectively. The above results were encouraging and worth pursuing for further development of Streptomyces griseoluteus GDJ1 (AT-R-S1) as an alternative resource for therapeutic L-asparaginase.

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Asparagine synthetase as a causal, predictive biomarker for l-asparaginase activity in ovarian cancer cells

Cited by 94 publications

References 46 publications

Asparagine synthetase expression and its potential prognostic value in patients with NK/T cell lymphoma

Asparagine synthetase expression and its potential prognostic value in patients with NK/T cell lymphoma

The glutaminase activity of l-asparaginase is not required for anticancer activity against ASNS-negative cells

Purification and Cytotoxicity of L-asparaginase from Streptomyces griseoluteus GDJ1

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