Thanks, Tuck & WS,
Appreciate the feedback. Here are the latest abstracts that my "siRNA" Google alert has pulled up. The first includes mention of a private siRNA company called Intradigm (www.intradigm).
Cheers,
Thomas
ajp.amjpathol.org
Inhibition of Ocular Angiogenesis by siRNA Targeting Vascular Endothelial Growth Factor Pathway Genes
Therapeutic Strategy for Herpetic Stromal Keratitis
Bumseok Kim*, Qingquan Tang, Partha S. Biswas*, Jun Xu, Raymond M. Schiffelers, Frank Y. Xie, Aslam M. Ansari, Puthupparampil V. Scaria, Martin C. Woodle, Patrick Lu and Barry T. Rouse*
From Comparative and Experimental Medicine,* College of Veterinary Medicine, University of Tennessee, Knoxville, Tennessee; and the Department of Genomics and Therapeutics, Intradigm Corporation, Rockville, Maryland
Ocular neovascularization often results in vision impairment. Frequently vascular endothelial cell growth factors (VEGFs) are mainly responsible for the pathological neovascularization as in the case in neovascularization induced by CpG oligodeoxynucleotides and herpes simplex virus infection in this report. siRNAs targeting either VEGFA, VEGFR1, VEGFR2, or a mix of the three were shown to significantly inhibit neovascularization induced by CpG when given locally or systemically. The efficacy of systemic administration was facilitated by the use of a polymer delivery vehicle. Additional experiments showed a significant inhibitory effect of the siRNAs mix when given either locally or systemically in vehicle against herpes simplex virus-induced angiogenesis as well as against lesions of stromal keratitis. These results indicate that the use of VEGF pathway-specific siRNAs represents a useful therapy against neovascularization-related eye diseases.
ajp.amjpathol.org
Role of Cks1 Overexpression in Oral Squamous Cell Carcinomas
Cooperation with Skp2 in Promoting p27 Degradation
Shojiro Kitajima*, Yasusei Kudo*, Ikuko Ogawa, Tarig Bashir, Masae Kitagawa*, Mutsumi Miyauchi*, Michele Pagano and Takashi Takata*
From the Department of Oral Maxillofacial Pathobiology,* Division of Frontier Medical Science, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan; the Center of Oral Clinical Examination, Hiroshima University Hospital, Hiroshima, Japan; and the Department of Pathology and NYU Cancer Institute, New York University School of Medicine, New York, New York
Down-regulation of p27 is frequently observed in various cancers due to an enhancement of its degradation. Skp2 is required for the ubiquitination and consequent degradation of p27 protein. Another protein called Cks1 is also required for p27 ubiquitination in the SCFSkp2 ubiquitinating machinery. In the present study, we examined Cks1 expression and its correlation with p27 in oral squamous cell carcinoma (OSCC) derived from tongue and gingiva. By immunohistochemical analysis, high expression of Cks1 was present in 62% of OSCCs in comparison with 0% of normal mucosae. In addition, 65% of samples with low p27 expression displayed high Cks1 levels. Finally, Cks1 expression was well correlated with Skp2 expression and poor prognosis. To study the role of Cks1 overexpression in p27 down-regulation, we transfected Cks1 with or without Skp2 into OSCC cells. Cks1 transfection could not induce a p27 down-regulation by itself, but both Cks1 and Skp2 transfection strongly induced. Moreover, we inhibited Cks1 expression by small interference RNA (siRNA) in OSCC. Cks1 siRNA transfection induced p27 accumulation and inhibited the growth of OSCC cells. These findings suggest that Cks1 overexpression may play an important role for OSCC development through Skp2-mediated p27 degradation, and that Cks1 siRNA can be a novel modality of gene therapy.
ajp.amjpathol.org
E4F1, a Novel Estrogen-Responsive Gene in Possible Atheroprotection, Revealed by Microarray Analysis
Yasuhiro Nakamura*, Katsuhide Igarashi, Takashi Suzuki*, Jun Kanno, Tohru Inoue, Chika Tazawa*, Masayuki Saruta*, Tomoko Ando, Noriko Moriyama, Toru Furukawa¶, Masao Ono*, Takuya Moriya*, Kiyoshi Ito||, Haruo Saito, Tadashi Ishibashi, Shoki Takahashi, Shogo Yamada and Hironobu Sasano*
From the Departments of Pathology,* Radiology, Molecular Pathology,¶ and Gynecology,|| Tohoku University School of Medicine, Sendai; and the Division of Toxicology and the Biological Safety Research Center, National Institute of Health Sciences, Tokyo, Japan
Estrogen has been postulated to be involved in inhibition of vascular smooth muscle cell (VSMC) proliferation mainly via estrogen receptor (ER), but the detailed mechanism has remained primarily unknown. Therefore, in this study, microarray analysis was used in two types of cultured human VSMCs: one positive for ER, and the other for ERß, which were treated by estrogens to detect the estrogen-responsive genes. We also used quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) to evaluate mRNA levels of selective target gene (TG) in these cells. We further studied whether the TG product was involved in inhibition of proliferation using small interfering RNA (siRNA) of the TG transfection. We subsequently used quantitative RT-PCR and in situ hybridization analysis to evaluate the expression of these gene products in human aorta. E4F1, a possible inducer of cell growth arrest, was markedly increased only in ER-positive VSMCs by estrogens in both microarray and RT-PCR analyses. Blocking of E4F1 using siRNA suppressed estrogenic inhibition of ER-positive VSMC proliferation. E4F1 mRNA was abundant in premenopausal female aorta with mild atherosclerotic changes. E4F1 is therefore considered one of the estrogen-responsive genes involving ER-mediated inhibition of VSMC proliferation and may play an important role in estrogen-related atheroprotection of human aorta. |
