Agonists of Toll-like Receptor 9 Containing Synthetic Dinucleotide Motifs

Yu, Dong; Putta, Mallikarjuna Reddy; Bhagat, Lakshmi; Li, Yukui; Zhu, Fu‐Gang; Wang, Daqing; Tang, Jimmy X.; Kandimalla, Ekambar R.; Agrawal, Sudhir

doi:10.1021/jm070881l

Cited by 24 publications

(24 citation statements)

References 32 publications

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“…11 Our data do not differentiate between sensing of ss or double-stranded DNA by TLR9 in endosomes. It is known that TLR9 signaling is stimulated by both double-stranded and ss unmethylated DNA molecules, [43][44][45][46][47] although sc (containing double 5Ј-end, sense-antisense sequences) TLR9 agonists induce more robust immune stimulation compared with single 5Ј-end cytosine guanine dinucleotide oligonucleotides. 48 It is feasible that the capsids of scAAV vectors are less stable in the endosomes and that thus more genomes are released and made available for TLR9 sensing as remains to be studied.…”

Section: Self-complementary Genomes Heighten Innate Immunity To Aav Vmentioning

confidence: 99%

The genome of self-complementary adeno-associated viral vectors increases Toll-like receptor 9–dependent innate immune responses in the liver

Martino

Suzuki

Markusic

et al. 2011

Blood

193

240

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IntroductionAdeno-associated viral (AAV) vectors are widely used for stable in vivo gene transfer to terminally differentiated or quiescent cells such as muscle fibers, hepatocytes, neurons, retinal cells, and others. These vectors, derived from a nonpathogenic, replicationdefective parvovirus with a small single-stranded (ss) DNA genome, have recently been successfully used in clinical gene transfer for inherited blindness and also show promise for other diseases. 1,2 Eight years ago, Zaiss et al 3 found that ssAAV serotype-2 vectors caused only a weak and highly transient innate immune response in the liver, suggesting that inflammatory reactions to AAV are negligible. Numerous animal studies have shown stable correction of genetic diseases by hepatic AAV gene transfer that may, in part, be because of the low innate immune profile of the vector, avoiding inflammatory signals. 4 In humans, hepatic gene transfer with AAV2 has been hampered by pre-existing adaptive immunity after natural infection in the form of neutralizing antibodies and capsid-specific CD8 ϩ T cells. 5 Numerous changes to capsid and vector genomes have been developed in recent years in attempts to improve gene transfer efficacy and possibly evade immunity. For example, AAV8 shows substantially higher transduction efficiency in mouse liver and reduced activation of capsid-specific T cells, and it facilitates tolerance induction to transgenes. 6,7 Furthermore, prevalence for neutralizing antibodies in humans is markedly lower to AAV8 than to AAV2. 8 In another set of investigations, replacing surfaceexposed tyrosine residues to phenylalanine has been shown to improve gene transfer for several serotypes. The resulting reduction in capsid phosphorylation in turn reduces accumulation in the cytoplasm (in favor of trafficking to the nucleus) and ubiquitination of capsid. 9 AAV2 gene transfer to hepatocytes was most improved by a combination of 3 Tyr-Phe changes in amino acid residues 444, 500, and 730. 10 Modifications of the recombinant AAV genome also can improve transduction rates. Being ss, the ssAAV genome has to be converted to a double-stranded form in the nucleus of an infected cell for transgene expression to occur. To overcome this ratelimiting step, self-complementary (sc)AAV vectors were developed by elimination of the terminal resolution site in one of the inverted terminal repeats (ITRs). 11 For such a genome to be packaged into capsid, the size of the expression cassette has to be further reduced to not exceed the packaging limit. Two groups reported optimized scAAV vectors for treatment of the X-linked bleeding disorder hemophilia B (coagulation factor IX deficiency) by liver gene transfer. 12,13 The hepatic microenvironment is more tolerogenic than that of many other tissues. 14 For example, we were able to tolerize hemophilia B mice to human factor IX (hF.IX) by hepatic ssAAV2 gene transfer. This protocol was successful in several strains, with the exception of C3H. 15 Nonetheless, AAV8 and AAV2(Y444/500/ 730F) vectors were able to...

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Section: Self-complementary Genomes Heighten Innate Immunity To Aav Vmentioning

confidence: 99%

The genome of self-complementary adeno-associated viral vectors increases Toll-like receptor 9–dependent innate immune responses in the liver

Martino

Suzuki

Markusic

et al. 2011

Blood

193

240

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“…We, therefore, hypothesized that systemic administration of TLR9 ligands could activate TLR signaling in the intestine and confer radioprotection and/or mitigation from RIGS. In this study we have used a novel TLR9 agonist containing synthetic immunomodulatory CpR (R = 2′-deoxy-7-dezaguanosine) dinucleotide and 3′-3′-attached novel structures that has been shown to induce potent TLR9-mediated immune responses [18], [19], [20], [21]. The presence of 3′-3′-attached structure provides higher metabolic stability and also optimal 5′-end presentation required for TLR9 recognition [18], [22], [23], [24].…”

Section: Introductionmentioning

confidence: 99%

TLR9 Agonist Protects Mice from Radiation-Induced Gastrointestinal Syndrome

et al. 2012

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PurposeRadiation-induced gastrointestinal syndrome (RIGS) is due to the clonogenic loss of crypt cells and villi depopulation, resulting in disruption of mucosal barrier, bacterial invasion, inflammation and sepsis. Intestinal macrophages could recognize invading bacterial DNA via TLR9 receptors and transmit regenerative signals to the neighboring crypt. We therefore investigated whether systemic administration of designer TLR9 agonist could ameliorate RIGS by activating TLR9.Methods and MaterialsMale C57Bl6 mice were distributed in four experimental cohorts, whole body irradiation (WBI) (8.4–10.4 Gy), TLR9 agonist (1 mg/kg s.c.), 1 h pre- or post-WBI and TLR9 agonist+WBI+iMyd88 (pretreatment with inhibitory peptide against Myd88). Animals were observed for survival and intestine was harvested for histological analysis. BALB/c mice with CT26 colon tumors in abdominal wall were irradiated with 14 Gy single dose of whole abdominal irradiation (AIR) for tumor growth study.ResultsMice receiving pre-WBI TLR9 agonist demonstrated improvement of survival after 10.4 Gy (p<0.03), 9.4 Gy (p<0.008) and 8.4 Gy (p<0.002) of WBI, compared to untreated or iMyd88-treated controls. Post-WBI TLR9 agonist mitigates up to 8.4 Gy WBI (p<0.01). Histological analysis and xylose absorption test demonstrated significant structural and functional restitution of the intestine in WBI+TLR9 agonist cohorts. Although, AIR reduced tumor growth, all animals died within 12 days from RIGS. TLR9 agonist improved the survival of mice beyond 28 days post-AIR (p<0.008) with significant reduction of tumor growth (p<0.0001).ConclusionsTLR9 agonist treatment could serve both as a prophylactic or mitigating agent against acute radiation syndrome and also as an adjuvant therapy to increase the therapeutic ratio of abdominal Radiation Therapy for Gastro Intestinal malignancies.

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“…5,6,8,9,11 In the present study, we designed 21-mer ODNs containing CpG* (ODNs 1−4) immune-stimulatory dinucleotides (Table 1). Each ODN contains two immune-stimulatory dinucleotides, one at the 5′-end and the second near the 3′-end, referred to as 5′-and 3′-immune-stimulatory dinucleotides, respectively (Table 1).…”

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confidence: 99%

Immune-Stimulatory Dinucleotide at the 5′-End of Oligodeoxynucleotides Is Critical for TLR9-Mediated Immune Responses

Putta

Bhagat

Wang

et al. 2013

ACS Med. Chem. Lett.

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Oligodeoxynucleotides (ODNs) containing a CpG or certain synthetic dinucleotides, referred to as immune-stimulatory dinucleotides, induce Toll-like receptor 9 (TLR9)-mediated immune responses. Chemical modifications such as 2'-O-methylribonucleotides incorporated adjacent to the immune-stimulatory dinucleotide on the 5'-side abrogate TLR9-mediated immune responses. In this study, we evaluated the effect of the location of immune-stimulatory dinucleotides in ODNs on TLR9-mediated immune responses. We designed and synthesized ODNs with two immune-stimulatory dinucleotides, one placed toward the 5'-end region and the other toward the 3'-end region, incorporated 2'-O-methylribonucleotides selectively preceding the 5'- or 3'-immune-stimulatory dinucleotide or both, and studied TLR9-mediated immune responses of these compounds in cell-based assays and in vivo in mice. These studies showed that an immune-stimulatory dinucleotide located closer to the 5'-end is critical for and dictates TLR9-mediated immune responses. These studies provide insights for the use of ODNs when employed as TLR9 agonists and antagonists or antisense agents.

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Agonists of Toll-like Receptor 9 Containing Synthetic Dinucleotide Motifs

Cited by 24 publications

References 32 publications

The genome of self-complementary adeno-associated viral vectors increases Toll-like receptor 9–dependent innate immune responses in the liver

The genome of self-complementary adeno-associated viral vectors increases Toll-like receptor 9–dependent innate immune responses in the liver

TLR9 Agonist Protects Mice from Radiation-Induced Gastrointestinal Syndrome

Immune-Stimulatory Dinucleotide at the 5′-End of Oligodeoxynucleotides Is Critical for TLR9-Mediated Immune Responses

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