Effect of G-rich sequences on the synthesis, purification, hybridization, cell uptake, and hemolytic activity of oligonucleotides

Agrawal, Sudhir; Iadarola, Patricia L.; Temsamani, Jamal; Zhao, Qiuyan; Shaw, Denise R.

doi:10.1016/0960-894x(96)00403-9

Cited by 36 publications

(17 citation statements)

References 9 publications

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“…In contrast, the cellular uptake of non-quadruplex sequences was negligible. Several groups have noted that the inclusion of polyguanine runs at the ends of PS, PO and mixed PS/PO oligonucleotides can improve cellular uptake [34,75-81,83,88-90]. For example, addition of a G 10 sequence to the 3′-end of an antisense oligodeoxynucleotide increased its uptake in J774E macrophage cells by 10-fold compared to the unmodified or 3′-C 10 oligonucleotide [89].…”

Section: Unusual Biological Properties Of G-rich Oligonucleotidesmentioning

confidence: 99%

Discovery and development of the G-rich oligonucleotide AS1411 as a novel treatment for cancer

Bates

Laber

Miller

et al. 2009

Experimental and Molecular Pathology

723

602

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Certain guanine-rich (G-rich) DNA and RNA molecules can associate intermolecularly or intramolecularly to form four stranded or "quadruplex" structures, which have unusual biophysical and biological properties. Several synthetic G-rich quadruplex-forming oligodeoxynucleotides have recently been investigated as therapeutic agents for various human diseases. We refer to these biologically active G-rich oligonucleotides as aptamers because their activities arise from binding to protein targets via shape-specific recognition (analogous to antibody-antigen binding). As therapeutic agents, the G-rich aptamers may have some advantages over monoclonal antibodies and other oligonucleotide-based approaches. For example, quadruplex oligonucleotides are nonimmunogenic, heat stable and they have increased resistance to serum nucleases and enhanced cellular uptake compared to unstructured sequences. In this review, we describe the characteristics and activities of G-rich oligonucleotides. We also give a personal perspective on the discovery and development of AS1411, an antiproliferative G-rich phosphodiester oligonucleotide that is currently being tested as an anticancer agent in Phase II clinical trials. This molecule functions as an aptamer to nucleolin, a multifunctional protein that is highly expressed by cancer cells, both intracellularly and on the cell surface. Thus, the serendipitous discovery of the G-rich oligonucleotides also led to the identification of nucleolin as a new molecular target for cancer therapy. KeywordsAS1411; aptamer; G-rich oligonucleotides; quadruplex; G-quartets; nucleolin; NF-kappaB; PRMT5; T-oligos; Dz13 Oligonucleotides as Therapeutic AgentsSince automated DNA synthesizers became widely available in the 1980s, there has been substantial interest in developing synthetic oligonucleotides for use as therapeutic agents [1][2][3][4][5][6][7][8][9]. Initial strategies aimed to prevent translation or transcription of specific viral or cellular * To whom correspondence should be addressed: 580 S. Preston St., Louisville, Kentucky 40202; Phone: (502) 852 2432; Email: E-mail: paula.bates@louisville.edu. 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 during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Conflicts of Interest Statement:Some of the authors (PJB, JOT, DMM) are inventors of patented or patent-pending technologies related to AS1411, G-rich oligonucleotides and nucleolin. Some of the authors (PJB, JOT, DMM) are shareholders in Antisoma, the company that is now sponsoring the development of AS1411. In addition, PJB is presently a recipient of grant support from Antisoma on projects related to AS14...

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Section: Unusual Biological Properties Of G-rich Oligonucleotidesmentioning

confidence: 99%

Discovery and development of the G-rich oligonucleotide AS1411 as a novel treatment for cancer

Bates

Laber

Miller

et al. 2009

Experimental and Molecular Pathology

723

602

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“…PS-ODN stock solutions were prepared just before their use and added to mycobacterial cultures after sterilization through 0.45-m HT Tuffryn membrane filters (Gelman). Poly(G) and 2Ј-O-methyl modifications of PS-ODNs were synthesized as described (13)(14)(15)(16).…”

Section: Methodsmentioning

confidence: 99%

“…(ii) The PS-ODN complementary to the 5Ј terminal 24 aa-encoding bases of the 32A-kDa gene transcript was extended at the 3Ј end beyond its standard length of 24 bases by 5 additional nonhybridizing G nucleotides. The first modification promotes PS-ODN stability and diminishes animal toxicity (14,15). The second modification was postulated to enhance uptake of the PS-ODN based on evidence from eukaryotic systems, showing increased uptake of PS-ODNs modified by the terminal G residues (16).…”

Section: Effect On M Tuberculosis Proliferation Of Modified Ps-odns mentioning

confidence: 99%

Targeting the Mycobacterium tuberculosis 30/32-kDa mycolyl transferase complex as a therapeutic strategy against tuberculosis: Proof of principle by using antisense technology

Harth

Horwitz

Tabatadze

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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We have investigated the effect of sequence-specific antisense phosphorothioate-modified oligodeoxyribonucleotides (PS-ODNs) targeting different regions of each of the 30͞32-kDa protein complex (antigen 85 complex) encoding genes on the multiplication of Mycobacterium tuberculosis. Single PS-ODNs to one of the three mycolyl transferase transcripts, added either once or weekly over the 6-wk observation period, inhibited bacterial growth by up to 1 log unit. A combination of three PS-ODNs specifically targeting all three transcripts inhibited bacterial growth by Ϸ2 logs; the addition of these PS-ODNs weekly for 6 wk was somewhat more effective than a one-time addition. Targeting the 5 end of the transcripts was more inhibitory than targeting internal sites; the most effective PS-ODNs and target sites had minimal or no secondary structure. The effect of the PS-ODNs was specific, as mismatched PS-ODNs had little or no inhibitory activity. The antisense PS-ODNs, which were highly stable in M. tuberculosis cultures, specifically blocked protein expression by their gene target. PS-ODNs targeting the transcript of a related 24-kDa protein (mpt51) had little inhibitory effect by themselves and did not increase the effect of PS-ODNs against the three members of the 30͞32-kDa protein complex. The addition of PS-ODNs against the transcripts of glutamine synthetase I (glnA1) and alanine racemase (alr) modestly increased the inhibitory efficacy of the 30͞32-kDa protein complex-specific PS-ODNs to Ϸ2.5 logs. This study shows that the three mycolyl transferases are highly promising targets for antituberculous therapy by using antisense or other antimicrobial technologies.phosphorothioate oligodeoxyribonucleotides ͉ mRNA secondary structure ͉ inhibition of translation initiation ͉ drug design T uberculosis, caused primarily by the facultative intracellular bacterium Mycobacterium tuberculosis, is the leading cause of death from a single infectious agent. The emergence of multidrug-resistant strains of M. tuberculosis has given urgency to the need for novel antimicrobial agents to combat this pathogen (1).M. tuberculosis expresses and secretes three closely related mycolyl transferases of 30-32 kDa mass, also known as the antigen 85 protein complex (antigen 85A, 85B, and 85C are referred to herein as the 32A, 30, and 32B proteins, respectively). All three enzymes catalyze the transfer of mycolic acid from one trehalose 6-monomycolate to another, resulting in trehalose 6,6Ј-dimycolate and free trehalose (2). The 3D structure of two of the enzymes, the 30 and 32A proteins, their catalytic sites, and their reaction mechanisms have been described in detail (3, 4). All three proteins are abundantly secreted. The 30-kDa protein is the most abundant extracellular protein of M. tuberculosis, responsible for nearly one quarter of the total extracellular protein in broth culture, and the 32A-kDa protein is the second most abundant extracellular protein (5). The three mycolyl transferases are also abundantly expressed in human macrophages i...

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“…Presumably, G-free SAT05f might have advantageous attributes over other inhibitory ODNs. ODNs with contiguous Gs show pronounced polyanionic related side-effects exemplified by prolongation of partial thromboplastin time [18,19]. ODNs with G-quadruplexes form unpredictable hyperstructure that facilitates the recognition by cellular proteins, thus leading to cytotoxicity in some cases [20].…”

Section: Contents Lists Available At Sciverse Sciencedirectmentioning

confidence: 99%

Structure–activity relationship of a guanine-free oligodeoxynucleotide as immunopotent inhibitor

Zhang

Wang

et al. 2012

International Immunopharmacology

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Effect of G-rich sequences on the synthesis, purification, hybridization, cell uptake, and hemolytic activity of oligonucleotides

Cited by 36 publications

References 9 publications

Discovery and development of the G-rich oligonucleotide AS1411 as a novel treatment for cancer

Discovery and development of the G-rich oligonucleotide AS1411 as a novel treatment for cancer

Targeting the Mycobacterium tuberculosis 30/32-kDa mycolyl transferase complex as a therapeutic strategy against tuberculosis: Proof of principle by using antisense technology

Structure–activity relationship of a guanine-free oligodeoxynucleotide as immunopotent inhibitor

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