PEGylation increases antitumoral activity of arginine deiminase of Streptococcus pyogenes

Abstract Arginine auxotrophy is a metabolic defect that renders tumor cells vulnerable towards arginine-depleting substances, such as arginine deiminase (ADI) from Streptococcus pyogenes (SpyADI). Previously, we confirmed SpyADI susceptibility on patient-derived glioblastoma multiforme (GBM) models in vitro and in vivo. For application in patients, serum half-life of the enzyme has to be increased and immunogenicity needs to be reduced. For this purpose, we conjugated the S. pyogenes-derived SpyADI with 20 kDa polyethylene glycol (PEG20) moieties, achieving a PEGylation of seven to eight of the 26 accessible primary amines of the SpyADI. The PEGylation reduced the overall activity of the enzyme by about 50% without affecting the Michaelis constant for arginine. PEGylation did not increase serum stability of SpyADI in vitro, but led to a longer-lasting reduction of plasma arginine levels in mice. Furthermore, SpyADI-PEG20 showed a higher antitumoral capacity towards GBM cells in vitro than the native enzyme. Key points • PEGylation has no effect on the affinity of SpyADI for arginine • PEGylation increases the antitumoral effects of SpyADI on GBM in vitro • PEGylation prolongs plasma arginine depletion by SpyADI in mice

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Arginine deprivation using pegylated arginine deiminase has activity against primary acute myeloid leukemia cells in vivo

Blood ◽

10.1182/blood-2014-10-608133 ◽

2015 ◽

Vol 125 (26) ◽

pp. 4060-4068 ◽

Cited By ~ 46

Author(s):

Farideh Miraki-Moud ◽

Essam Ghazaly ◽

Linda Ariza-McNaughton ◽

Katharine A. Hodby ◽

Andrew Clear ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Arginine Deiminase ◽

Leukemia Cells ◽

Arginine Deprivation ◽

Key Points ◽

Acute Myeloid Leukemia Cells ◽

Acute Myeloid

Key Points Most AMLs lack ASS1, which allows synthesis of arginine, and so depend on exogenous sources. Depletion of arginine via ADI-PEG 20 reduces the burden of primary AML in vivo and in vitro.

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Streptococcus pyogenes Arginine and Citrulline Catabolism Promotes Infection and Modulates Innate Immunity

Infection and Immunity ◽

10.1128/iai.00916-13 ◽

2013 ◽

Vol 82 (1) ◽

pp. 233-242 ◽

Cited By ~ 24

Author(s):

Zachary T. Cusumano ◽

Michael E. Watson ◽

Michael G. Caparon

Keyword(s):

Nitric Oxide ◽

Innate Immunity ◽

Streptococcus Pyogenes ◽

Murine Model ◽

Acid Stress ◽

Arginine Deiminase ◽

Content Type ◽

Cutaneous Tissue

ABSTRACTA bacterium's ability to acquire nutrients from its host during infection is an essential component of pathogenesis. For the Gram-positive pathogenStreptococcus pyogenes, catabolism of the amino acid arginine via the arginine deiminase (ADI) pathway supplements energy production and provides protection against acid stressin vitro. Its expression is enhanced in murine models of infection, suggesting an important rolein vivo. To gain insight into the function of the ADI pathway in pathogenesis, the virulence of mutants defective in each of its enzymes was examined. Mutants unable to use arginine (ΔArcA) or citrulline (ΔArcB) were attenuated for carriage in a murine model of asymptomatic mucosal colonization. However, in a murine model of inflammatory infection of cutaneous tissue, the ΔArcA mutant was attenuated but the ΔArcB mutant was hyperattenuated, revealing an unexpected tissue-specific role for citrulline metabolism in pathogenesis. When mice defective for the arginine-dependent production of nitric oxide (iNOS−/−) were infected with the ΔArcA mutant, cutaneous virulence was rescued, demonstrating that the ability ofS. pyogenesto utilize arginine was dispensable in the absence of nitric oxide-mediated innate immunity. This work demonstrates the importance of arginine and citrulline catabolism and suggests a novel mechanism of virulence by whichS. pyogenesuses its metabolism to modulate innate immunity through depletion of an essential host nutrient.

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Faculty Opinions recommendation of A fluoroacetamidine-based inactivator of protein arginine deiminase 4: design, synthesis, and in vitro and in vivo evaluation.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1031506.367705 ◽

2006 ◽

Author(s):

Karen Allen

Keyword(s):

Arginine Deiminase ◽

In Vivo Evaluation ◽

Design Synthesis ◽

Protein Arginine Deiminase

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The Design and Preclinical Evaluation of a Single-Label Bimodal Nanobody Tracer for Image-Guided Surgery

Biomolecules ◽

10.3390/biom11030360 ◽

2021 ◽

Vol 11 (3) ◽

pp. 360

Author(s):

Pieterjan Debie ◽

Noemi B. Declerck ◽

Danny van Willigen ◽

Celine M. Huygen ◽

Bieke De Sloovere ◽

...

Keyword(s):

Single Molecule ◽

Gamma Ray ◽

Nuclear Imaging ◽

Primary Amines ◽

Resection Margins ◽

Fluorescent Light ◽

Single Structure ◽

Fluorescence Images

Intraoperative guidance using targeted fluorescent tracers can potentially provide surgeons with real-time feedback on the presence of tumor tissue in resection margins. To overcome the limited depth penetration of fluorescent light, combining fluorescence with SPECT/CT imaging and/or gamma-ray tracing has been proposed. Here, we describe the design and preclinical validation of a novel bimodal nanobody-tracer, labeled using a “multifunctional single attachment point” (MSAP) label, integrating a Cy5 fluorophore and a diethylenetriaminepentaacetic acid (DTPA) chelator into a single structure. After conjugation of the bimodal MSAP to primary amines of the anti-HER2 nanobody 2Rs15d and 111In-labeling of DTPA, the tracer’s characteristics were evaluated in vitro. Subsequently, its biodistribution and tumor targeting were assessed by SPECT/CT and fluorescence imaging over 24 h. Finally, the tracer’s ability to identify small, disseminated tumor lesions was investigated in mice bearing HER2-overexpressing SKOV3.IP1 peritoneal lesions. [111In]In-MSAP.2Rs15d retained its affinity following conjugation and remained stable for 24 h. In vivo SPECT/CT and fluorescence images showed specific uptake in HER2-overexpressing tumors with low background. High tumor-to-muscle ratios were obtained at 1h p.i. and remained 19-fold on SPECT/CT and 3-fold on fluorescence images over 24 h. In the intraperitoneally disseminated model, the tracer allowed detection of larger lesions via nuclear imaging, while fluorescence enabled accurate removal of submillimeter lesions. Bimodal nuclear/fluorescent nanobody-tracers can thus be conveniently designed by conjugation of a single-molecule MSAP-reagent carrying a fluorophore and chelator for radioactive labeling. Such tracers hold promise for clinical applications.

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The Transcriptional Regulator CpsY Is Important for Innate Immune Evasion in Streptococcus pyogenes

Infection and Immunity ◽

10.1128/iai.00925-16 ◽

2016 ◽

Vol 85 (3) ◽

Cited By ~ 2

Author(s):

Luis A. Vega ◽

Kayla M. Valdes ◽

Ganesh S. Sundar ◽

Ashton T. Belew ◽

Emrul Islam ◽

...

Keyword(s):

Streptococcus Pyogenes ◽

Immune Evasion ◽

Immune Cell ◽

Group A Streptococcus ◽

Transcriptional Regulator ◽

Innate Immune ◽

Content Type ◽

Human Host

ABSTRACTAs an exclusively human pathogen,Streptococcus pyogenes(the group A streptococcus [GAS]) has specifically adapted to evade host innate immunity and survive in multiple tissue niches, including blood. GAS can overcome the metabolic constraints of the blood environment and expresses various immunomodulatory factors necessary for survival and immune cell resistance. Here we present our investigation of one such factor, the predicted LysR family transcriptional regulator CpsY. The encoding gene,cpsY, was initially identified as being required for GAS survival in a transposon-site hybridization (TraSH) screen in whole human blood. CpsY is homologous with transcriptional regulators ofStreptococcus mutans(MetR),Streptococcus iniae(CpsY), andStreptococcus agalactiae(MtaR) that regulate methionine transport, amino acid metabolism, resistance to neutrophil-mediated killing, and survivalin vivo. Our investigation indicated that CpsY is involved in GAS resistance to innate immune cells of its human host. However, GAS CpsY does not manifest thein vitrophenotypes of its homologs in other streptococcal species. GAS CpsY appears to regulate a small set of genes that is markedly different from the regulons of its homologs. The differential expression of these genes depends on the growth medium, and CpsY modestly influences their expression. The GAS CpsY regulon includes known virulence factors (mntE,speB,spd,nga[spn],prtS[SpyCEP], andsse) and cell surface-associated factors of GAS (emm1,mur1.2,sibA[cdhA], andM5005_Spy0500). Intriguingly, the loss of CpsY in GAS does not result in virulence defects in murine models of infection, suggesting that CpsY function in immune evasion is specific to the human host.

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Bmk9 and Uricase Nanoparticle Complex for the Treatment of Gouty Arthritis and Uric Acid Nephropathy

Journal of Biomedical Nanotechnology ◽

10.1166/jbn.2021.3168 ◽

2021 ◽

Vol 17 (10) ◽

pp. 2071-2084

Author(s):

Tianjiao Han ◽

Meiying Wang ◽

Wenchao Li ◽

Mingxing An ◽

Hongzheng Fu

Keyword(s):

Uric Acid ◽

Drug Loading ◽

Gouty Arthritis ◽

The Body ◽

Key Points ◽

Uric Acid Nephropathy ◽

Charged Carrier ◽

Dual Drug

Uric acid is the final product of purine metabolism, and excessive serum uric acid can cause gouty arthritis and uric acid nephropathy. Therefore, lowering the uric acid level and alleviating inflammation in the body are the key points to treating these diseases. A stable nanosuspension of peptide BmK9 was prepared by the precipitation-ultrasonication method. By combining uricase on the surface of a positively charged carrier, a complex consisting of neutral rod-shaped BmK9 and uricase nanoparticles (Nplex) was formed to achieve the delivery of BmK9 and uricase, respectively. The formulation of Nplex has a diameter of 180 nm and drug loading up to 200%, which releases BmK9 and uricase slowly and steadily in drug release tests in vitro. There was significantly improved pharmacokinetic behavior of the two drugs because Nplex prolonged the half-life and increased tissue accumulation. Histological assessments showed that the dual drug Nplex can reduce the inflammation response in acute gouty arthritis and chronic uric acid nephropathy in vivo. In the macrophage system, there was lower toxicity and increased beneficial effect on inflammation with Nplex than free BmK9 or uricase. Collectively, this novel formulation provides a dual drug delivery system that can treat gouty arthritis and uric acid nephropathy.

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Chitin conduits modified with DNA-peptide coating promote the peripheral nerve regeneration

Biofabrication ◽

10.1088/1758-5090/ac3bdc ◽

2021 ◽

Author(s):

Songyang Liu ◽

Liping Zhou ◽

Ci Li ◽

Tiantian Min ◽

Changfeng Lu ◽

...

Keyword(s):

Peripheral Nerve ◽

Nerve Injury ◽

Peripheral Nerve Injury ◽

Amide Bond ◽

Primary Amines ◽

Direct Reaction ◽

Ct Dna ◽

Peptide Coating

Abstract Peripheral nerve injury (PNI) is one of the common clinical injuries which needs to be addressed. Previous studies demonstrated the effectiveness of using biodegradable chitin (CT) conduits small gap tubulization technology as a substitute for traditional epineurial neurorrhaphy. Aiming to improve the effectiveness of CT conduits in repairing PNI, we modified their surface with a DNA-peptide coating. The coating consisted of single strand DNA (ssDNA) and its complementary DNA’-peptide mimics. First, we immobilize ssDNA (DNA1+2) on CT conduits by EDC/NHS method to construct CT/DNA conduits. EDC/NHS was used to activate carboxyl groups of modified ssDNA for direct reaction with primary amines on the chitin via amide bond formation. Then, DNA1’-BDNF+DNA2’-VEGF mimic peptide （RGI+KLT）were bonded to CT/DNA conduits by complementary base pairing principle at room temperature to form CT/RGI+KLT conduits. When the surrounding environment rose to a certain point (37℃), the CT/RGI+KLT conduits achieved sustainable release of DNA’-peptide. In vitro, the CT conduits modified with the DNA-peptide coating promoted the proliferation and secretion of Schwann cells by maintaining their repair state. It also promoted the proliferation of HUVECs and axon outgrowth of DRG explants. In vivo, CT/RGI+KLT conduits promoted regeneration of injured nerves and functional recovery of target muscles, which was facilitated by the synergistic contribution of angiogenesis and neurogenesis. Our research brings DNA and DNA-peptide hybrids into the realm of tissue engineering to repair peripheral nerve injury.

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Therapeutic targeting of HES1 transcriptional programs in T-ALL

Blood ◽

10.1182/blood-2014-10-608448 ◽

2015 ◽

Vol 125 (18) ◽

pp. 2806-2814 ◽

Cited By ~ 31

Author(s):

Stephanie A. Schnell ◽

Alberto Ambesi-Impiombato ◽

Marta Sanchez-Martin ◽

Laura Belver ◽

Luyao Xu ◽

...

Keyword(s):

Therapeutic Targeting ◽

Key Points

Key Points NOTCH1 inhibits apoptosis via HES1-mediated repression of BBC3 in T-ALL. Perhexiline, a HES1 signature modulator drug, has strong antileukemic effects in vitro and in vivo.

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Dicer1-mediated miRNA processing shapes the mRNA profile and function of murine platelets

Blood ◽

10.1182/blood-2015-07-661371 ◽

2016 ◽

Vol 127 (14) ◽

pp. 1743-1751 ◽

Cited By ~ 48

Author(s):

Jesse W. Rowley ◽

Stéphane Chappaz ◽

Aurélie Corduan ◽

Mark M. W. Chong ◽

Robert Campbell ◽

...

Keyword(s):

Functional Responses ◽

Mirna Processing ◽

Key Points ◽

Mrna Profile ◽

And Function

Key Points Dicer1 deletion in MKs alters platelet miRNA and mRNA profiles. Dicer1-deficient platelets display increased integrins αIIb and β3 levels and enhanced in vitro and in vivo functional responses.

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