Arginine deprivation by arginine deiminase ofStreptococcus pyogenescontrols primary glioblastoma growthin vitroandin vivo

Abstract

Arginine auxotrophy constitutes a weak point of several tumors, among them glioblastoma multiforme (GBM). Hence, those tumors are supposed to be sensitive for arginine-depleting substances, such as arginine deiminase (ADI). Here we elucidated the sensitivity of patient-individual GBM cell lines toward Streptococcus pyogenes-derived ADI. To improve therapy, ADI was combined with currently established and pre-clinical cytostatic drugs. Additionally, effectiveness of local ADI therapy was determined in xenopatients. Half of the GBM cell lines tested responded well toward ADI monotherapy. In those cell lines, viability decreased significantly (up to 50 %). Responding cell lines were subjected to combination therapy experiments to test if any additive or even synergistic effects may be achieved. Such promising results were obtained in 2/3 cases. In cell lines HROG02, HROG05 and HROG10, ADI and Palomid 529 combinations were most effective yielding more than 70 % killing after 2 rounds of treatment. Comparable boosted antitumoral effects were observed after adding chloroquine to ADI (>60% killing). Apoptosis, as well as cell cycle dysregulation were found to play a minor role. In some, but clearly not all cases, (epi-) genetic silencing of arginine synthesis pathway genes (argininosuccinate synthetase 1 and argininosuccinate lyase) explained obtained results. In vivo, ADI as well as the combination of ADI and SAHA efficiently controlled HROG05 xenograft growth, whereas adding Palomid 529 to ADI did not further increase the strong antitumoral effect of ADI. The cumulative in vitro and in vivo results proved ADI as a very promising candidate therapeutic, especially for development of adjuvant GBM combination treatments.

Arginine-Depleting Enzymes – An Increasingly Recognized Treatment Strategy for Therapy-Refractory Malignancies

“…In a previous publication by Syed et al, 22 % of primary GBM cultures were found to harbor a methylated CpG island in the ASL core promoter, leading to significant downregulation of ASL mRNA [27]. In our own studies on patient-derived GBM cell lines, comparable results were obtained, with reduced ASL mRNA levels in half of the examined cases (12 in total) [30]. Additional genes OTC and arginase I were almost entirely downregulated in this cohort, whereas carbamoyl phosphate synthase I was expressed by all cell lines [31].…”

“…3). This is accompanied by cell cycle alterations (G 1 or S phase arrest), as proven for pancreatic and breast cancer as well as GBM cells [30,31,71].…”

“…Hence, arginine-degrading enzymes are currently the most promising approach to arginine deprivation therapy. Therapeutically relevant argininedegrading enzymes include (1) arginase 1 catabolizing arginine to ornithine, (2) arginine decarboxylase (ADC) decarboxylating arginine to agmatine and CO 2 , and (3) arginine deiminase (ADI) degrading arginine to citrulline and ammonia [19,30,46]. The potential of each of these enzymes for arginine degradation not only depends on their efficiency, but also on their immunogenicity, catabolic products as well as stability in patients.…”

Efficacy of Asparaginase Erwinia chrysanthemi With and Without Temozolomide Against Glioma Cells and Intracranial Mouse Medulloblastoma

“…The concept of amino acid depletion contributing to antibrain tumor activity may not be unique to Asn and Gln, because others have demonstrated similar anti-GBM effects with manipulation of arginine metabolism (38,39). Our study adds evidence justifying further research on amino acid manipulation for development of brain tumor therapeutics.…”

A Role of Arginine Deiminase From Streptococcus Pyogenes M49-16 in Promoting Infection and Inhibition of Endothelial Cell Proliferation

“…Была доказана способ-ность фермента, подавлять пролиферацию моно-нуклеарных лейкоцитов периферической крови, клеток глиобластомы, клеток карциномы кожи А431 и др. [9,13,21]. В то же время действие АД из S. pyogenes в отношении эндотелиальных кле-ток остается недостаточно изученным.…”