Molecular Human Reproduction, Vol. 9, No. 10, 603-609,
October 2003
© 2003 European Society of Human Reproduction and Embryology
Article |
Highly efficient and minimally invasive in-vivo gene transfer to the mouse uterus using haemagglutinating virus of Japan (HVJ) envelope vector
Submitted on May 7, 2003; accepted on June 26, 2003
1 Division of Obstetrics and Gynecology, Department of Specific Organ Regulation and 2 Division of Gene Therapy Science, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565–0871, Japan
3 To whom correspondence should be addressed. e-mail: tadashi{at}gyne.med.osaka-u.ac.jp
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and methodsResultsDiscussionReferences |
|---|
The uterus is obviously critical in implantation, development of the fetus and parturition. Endometrial cancer derived from endometrial epithelium is one of the common malignancies in the female reproductive tract. In order to clarify the local mechanisms of reproductive physiology and establish a non-systemic therapeutic strategy for reproductive failure as well as for endometrial cancer, we applied haemaggulutinating virus of Japan envelope (HVJ-E) vector to in-vivo gene transfer into the uterine cavity of IVCS mice. Injection of HVJ-E vector into mouse uterine cavity on day 1.5 post coitum (p.c.) introduced a reporter gene
120-fold more efficiently than introduction using the cationic liposome method. The expression of the introduced gene continued for at least 3 days. The plasmid vector was localized in the endometrial epithelium, whereas oligo deoxynucleotides were distributed throughout the epithelium, stromal cells and myometrium. HVJ-E vector did not affect the pregnancy rate, course of pregnancy, litter size, fetal growth in utero or parturition, and did not transfect the exogenous gene to the fetus. These results indicate that gene transfer into the uterus using HVJ-E vector is highly efficient and safe during pregnancy, and results in a well controlled distribution of the exogenous DNA. We believe that this procedure should be widely applicable for investigations of reproductive physiology as well as for methods of local gene therapy in the uterus. Key words: in vivo/gene transfer/HVJ/pregnancy/uterus
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
|---|
The murine uterus is a tubular organ that consists of endometrium, endometrial stroma, myometrium and serosa. During reproduction, each component plays important roles that depend on the sex steroid hormone milieu. Estrogen and progesterone induce morphological and functional alterations in the endometrium and its stroma. After ovulation, these hormones induce the uterine cavity to become receptive to the fertilized embryo during the implantation window period. Disturbed sex steroid hormone secretion can lead to endometrial dysfunction and menstrual problems. Unopposed estrogen exposure for long periods stimulates abnormal cell growth in the endometrium and eventually causes endometrial hyperplasia and endometrial cancer (Grady et al., 1995). Some of these hormonal effects are direct; however, most of the local actions of the sex steroids are mediated by growth factors or cytokines in an autocrine and/or paracrine manner (Lindhart et al., 2002). To elucidate the function of growth factors and cytokines in implantation, gene-targeting appears to be ineffective. Knock-out mice for leukaemia inhibitory factor (Stewart et al., 1992), Hoxa-10 (Satokata et al., 1995), cyclooxygenase-2 (Lim et al., 1997) and interleukine 11
receptor (Bilinski et al., 1998) genes show implantation failure. Many knock-out genes are lethal in early life, cause infertility because of underdevelopment of the reproduction system or show no alteration of phenotype with respect to reproduction. For elucidating the function of various factors involved in implantation, in-vivo gene transfer to the uterine cavity should be a powerful tool. This tool will be able to apply not only for overexpression of the gene by expression plasmid, but also for down-regulation of the gene by antisense oligonucleotides for the gene or by decoy for the transcription factor binding sites. Facilitation of implantation for infertility treatment, emergency contraception via blockage of implantation, treatment of dysfunctional endometrium and early-stage, non-invasive endometrial cancer should be attractive candidates for gene therapy localized in the uterine cavity in order to avoid systemic complications (Sharkey, 2000). In order to establish in-vivo gene transfer as a reliable method for investigation of reproductive physiology and gene therapy, the following conditions are necessary: (i) high efficiency of gene transfer into the target tissue; (ii) a transfer technique that does not interfere with the physiology of pregnancy; (iii) lack of transfer of the gene to the fetus. Previously, successful in-vivo gene transfer to the female reproductive tract (uterus and oviduct) using cationic liposomes (Lipofectamine®: Charnock-Jones et al., 1997; Relloso and Esponda, 1998; Bagot et al., 2000; Daftary and Taylor, 2001; DOTAP®: Zhu et al., 1998; GenePorterTM: Hsieh et al., 2002) has been reported. Cationic liposomes show excellent entrapment of negatively charged macromolecules such as DNA and high efficiency of transfection to in-vitro cultured cells, whose plasma membranes are negatively charged. However, for in-vivo gene transfer, the use of cationic lipids instead of anionic lipids as a liposome component dramatically reduced the transfection efficiency (Saeki et al., 1997). We report here a novel in-vivo gene transfer technique using a haemagglutinating virus of Japan (HVJ) viral envelope HVJ-E vector with an anionic lipid component (Kaneda et al., 2002) and introduction into the murine uterus. We demonstrate that this method transfers the gene to the uterus approximately 120 times more efficiently than a technique using cationic liposomes. We also show that gene transfer using injection of the HVJ-E vector into the uterine cavity in the early stages of pregnancy does not interfere with the course of pregnancy and does not cause transfer of the exogenous gene into the fetus in utero.
|
Materials and methods |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
|---|
Animals
We examined vaginal smears to determine the sexual phase (proestrus, estrus, metestrus and diestrus) of female IVCS mice (SLC, Shizuoka, Japan) aged 8–10 weeks. Virgin female mice in the estrus phase were bred with male IVCS mice. The morning when a vaginal plugging was first observed was designated as day 0.5 post coitum (p.c.). All animal experiments were performed according to the appropriate guidelines for animal use approved by the Institutional Animal Care and Use Committee of Osaka University Graduate School of Medicine.
DNA preparation
In our preliminary experiments, we tested pEB (Epstein–Barr virus replicon-based plasmid; Saeki et al., 1998), pAct-NII (chicken ß-actin promoter and enhancer driven; Saeki et al., 1997), pEBAct (a fusion plasmid of the above two plasmids; Saeki et al., 1998) and pcDNA3 (cytomegalovirus promoter driven; Invitrogen, San Diego, CA) plasmid vectors to test the luciferase cDNA expression and found that only pcDNA3 could effectively mediate expression of the luciferase protein in the mouse uterus in vivo (data not shown). Therefore, luciferase cDNA or ß-galactosidase cDNA was introduced into pcDNA3 at appropriate restriction sites (pcDNA-Luc and pcDNA-LacZ). Plasmid DNA was purified using a Qiagen column (Tokyo, Japan). FITC-labelled oligodeoxynucleotide (FITC-ODN; 20mer with random sequence) was purchased from Hokkaido System Science (Sapporo, Japan). Control transfections were performed using the plasmid without a reporter gene.
Preparation of HVJ-E vector
HVJ (also known as Sendai virus) was amplified as described previously (Saeki and Kaneda, 1998). Virus was inactivated by ß-propiolactone (0.0075–0.001%) treatment or by UV irradiation. In both cases, the virus preparation lost the ability to replicate. Aliquots of the virus (3x1010 particles/1.5 ml tube) were centrifuged (18 500 g, 15 min) at 4°C, and the viral pellet was stored at –20°C (Kaneda et al., 2002).
Inactivated virus suspension [10 000 haemagglutinating activity units (HAU)] was mixed with plasmid DNA or oligonucleotide (800 µg of DNA), 8 µl of 3% Triton/TE and balanced salt solution [BSS;10 mmol/l Tris–HCl (pH 7.5), 137 mmol/l NaCl and 5.4 mmol/l KCl] to yield a final volume of 100 µl. The mixture was centrifuged at 18 500 g for 15 min at 4°C. After the pellet was washed with 1 ml of human tubal fluid (HTF) medium (Nippon Medical & Chemical Instrument Co. Ltd, Osaka, Japan) to remove the detergent and unincorporated DNA, the envelope vector was suspended in 250 µl of HTF medium. Approximately 15–20% of DNA was incorporated into the vector. The HVJ-E vector was stored at 4°C until use. HVJ-E vector is also commercially available from Ishihara Sangyo Co. Ltd (Osaka, Japan).
Cationic liposome–DNA preparation
Cationic liposome–DNA suspension was prepared using Lipofectamine [a 3:1 (w/w) formulation of 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N-N-dimethyl-1-propanaminium trifluoroacetate and dioleoylphosphatidyl ethanolamine; Invitrogen]. In brief, plasmid DNA was mixed with 10 µl of Lipofectamine and the mixture was incubated for 15 min at room temperature (22°C). A final amount of 16 or 80 µg of DNA and 40 µg of liposome was obtained by dilution with 1x Dulbecco’s phosphate-buffered saline (PBS) to a total volume of 100 µl.
In-vivo gene transfer
Mice on day 1.5 p.c. were anaesthetized and subjected to laparotomy to expose the uterus. Twenty five microlitres of HVJ-E vector containing plasmid DNA or FITC-ODN, or 25 µl of cationic liposome–DNA preparation were injected slowly into the uterine cavity using a 30-gauge needle, and the cervix was clamped for 10 min. Then, the incision was closed to allow recovery of the mice.
Luciferase assay
The whole uterus transfected with pcDNA-Luc was excised, rinsed in ice-cold PBS, minced and homogenized in lysis buffer provided by Promega (Madison, WI). Luciferase activity was measured with a luciferase assay kit (Promega) (Saeki et al., 1997), and the protein concentration was determined using a Bio-Rad protein assay kit (Bio-Rad, Hercules, CA) according to the manufacturer’s instructions. Bioluminescence was detected in triplicate using a Lumat LB9507 (Berthold, Wildbad, Germany) using the standard protocol provided by the manufacturer. In the standard curve, the data was linearized by plotting the log of luciferase concentrations versus the log of the relative light units (RLU) from 10–10 g/l (1.7x10–15 mol/l) to 10–5 g/l (1.7x10–10 mol/l) of purified firefly luciferase (Roche, Mannheim, Germany).
Detection of transfected DNA
The uterus was removed 24 h after transfection with pcDNA-Lac Z, prefixed with 0.25% glutaraldehyde in 2% paraformaldehyde (PFA)/PBS at 4°C overnight and dehydrated with 30% sucrose/PBS at 4°C overnight. After dehydration, the uterus was rinsed with ice-cold PBS and embedded in O.C.T. compound (Miles Scientific, Elkhart, IN). ß-Galactosidase staining was performed on cryostat sections (5 µm) as follows. The sections were dried and then incubated with X-Gal staining solution [PBS containing 2 mmol/l MgCl2, 5 mmol/l K3Fe(CN)6, 5 mmol/l K4Fe(CN)6 and 1 mg/ml X-Gal; pH 7.4] at 37°C overnight.
The uterus transfected with FITC-ODN was removed on day 3.5 p.c. The uterus was prefixed with 4% PFA/PBS at 4°C overnight and dehydrated with 30% sucrose/PBS at 4°C overnight. After dehydration, the uterus was rinsed with ice-cold PBS and cut into cryostat sections (5 µm). The fluorescent signal was visualized by fluorescence microscopy. The sections were counter-stained by haematoxylin–eosin staining.
Immunohistochemistry
Uteri transfected with 5000 HAU of empty HVJ-E vector without plasmid were removed at 24, 48 or 72 h after transfection. The uterus was embedded in O.T.C. compound and frozen sections (5 µm) were fixed with 4% PFA/PBS at 4°C for 5 min. After fixation, sections were incubated with blocking buffer containing 10% goat serum. The primary antiserum, against anti-F protein, was made by immunizing a rabbit with purified HVJ-F protein (Miura et al., 1993). The antiserum was diluted 1:1000 with 1% BSA/PBS buffer and incubated with the sections at 4°C overnight, and then the sections were rinsed with PBS three times. The immunoreactivity was visualized by incubation with goat anti-rabbit antibody labelled with FITC at room temperature for 30 min.
Polymerase chain reaction
On day 10.5 p.c., the fetuses and uteri were removed from pcDNA-Luc-transfected mice. DNA was extracted using lysis buffer [150 mmol/l NaCl, 10 mmol/l Tris–HCl (pH 8.0), 10 mmol/l EDTA, 0.1% SDS and 100 µg/ml proteinase K (Nacalai Tesque, Kyoto, Japan)], and purified by phenol–chloroform extraction and ethanol precipitation. PCR was performed using rTaq polymerase (Toyobo, Osaka, Japan). The reaction was carried out using purified DNA (1 µg) or luciferase gene plasmid (1 pg) in the presence of rTaq polymerase (1 IU), 1x rTaq buffer, dNTP mixture (0.125 mmol/l) and 10 pmol each of T7 and SP6 primers (Invitrogen). Amplification was performed in a total volume of 20 µl for 35 cycles (denaturation at 96°C for 60 s, annealing at 55°C for 30 s and extension at 72°C for 60 s) using a thermal cycler (Perkin-Elmer 2400, Norwalk, CT, USA). Ten microlitres of the reaction mixture was electrophoresed on a 1.5% agarose gel, stained with ethidium bromide, and visualized by UV illumination.
Statistical evaluation of results
Statistical analysis was performed using the Mann–Whitney U-test, and differences with a P-value <0.05 were considered significant.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
|---|
Efficiency of in-vivo gene transfer to the uterus
HVJ-E vector entraps
15–20% of applied DNA under our conditions (Kaneda et al., 2002). This implies that we injected 16 µg of plasmid/horn using HVJ-E vector. Therefore, we decided to use less (exp-1, 4 µg/horn) or more (exp-2, 20 µg/horn) than this amount of plasmid for the Lipofectamine vector in order to compare the gene transfer efficiencies using the same volume of fluid (25 µl) for injection/uterine horn. After 24 h of transfection, the luciferase activity using HVJ-E vector was approximately 120 times higher than that obtained using Lipofectamine (P < 0.005) (Figure 1a). A 5-fold increase of the amount of plasmid had no effect on the level of luciferase activity using Lipofectamine (Figure 1a). Luciferase activity was detected in the uterus for at least 3 days after transfection with HVJ-E (Figure 1b). All of the assays were performed within the linear range of the standard curve. In order to examine the tissue distribution of the transfected cells within the uterus, we transfected pcDNA-LacZ or a 20mer of FITC-labelled ODN using HVJ-E. After 24 h of transfection, ß-galactosidase activity was detected mainly in the luminal and glandular endometrial epithelium (Figure 2a–d). Few stromal cells were stained with this procedure. Interestingly, when we transfected FITC-ODN (16 µg/horn) into the uterus using HVJ-E, the ODN was distributed not only in the endometrial epithelium, but also in the stromal cell layer and in some myometrial cells (Figure 2e and f).
|
|
Safety of HVJ-E transfection during pregnancy
At first, a total of 18 mice were injected with HVJ-E vector in the uterus on day 1.5 p.c. The pregnancy rate was 100%, and all of the litters were delivered between day 18.5 and 19.5 p.c. (data not shown). We then compared in detail the course of pregnancy and outcome of the fetuses in mice injected in the uterus with empty HVJ-E vector (n = 5) or HTF (vehicle) (n = 5). In both groups, 100% of mice became pregnant, and the increase of maternal body weight on day 18.5 p.c. showed no significant difference. In both groups, parturition occurred between days 18.5 and 19.5 p.c. and no dystocia was observed. Litter size per mouse, birth weight and cranio-rump length of the pups also showed no significant difference (Table I). These data suggest that the injection of HVJ-E vector into the uterine cavity does not affect the uterine physiology during the course of pregnancy.
|
Rapid elimination of HVJ-E protein from the uterus
We examined the presence of HVJ-derived protein after injection by immunohistochemistry using FITC-labelled secondary antibody against anti-HVJ F-protein antibody. Twenty four hours and 48 h after injection, significant immunoreactivity of F-protein was observed in the endometrium, stroma and myometrium (Figure 3a and b). Even when we introduced a 5-fold larger amount of empty HVJ-E vector into the uterine horn, the immunoreactivity of F-protein disappeared after 72 h of transfection (Figure 3c). From this time course, we speculate that at the time of implantation (day 4.5–5.5 p.c.), the viral protein that is necessary to introduce the exogenous DNA into tissues has already been eliminated from the uterus. Haematoxylin–eosin staining of sections (Figure 3d) obtained after 72 h of transfection indicated the localization of HVJ-E vector.
|
Transfer of the exogenous DNA to fetuses
On day 10.5 p.c., we tested for the presence of transfected DNA in the uterus as well as in the fetus. After transfection of pcDNA-Luc with the HVJ-E vector, we did not detect the amplicon of luciferase cDNA in either the fetus or the uterus using 35 cycles of PCR (Figure 4).
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
|---|
Our in-vivo gene transfer system using HVJ-E vector showed
120-fold higher gene transfer efficiency than the method using cationic liposomes in our hands. We were at first concerned that a local immune reaction against the viral envelope protein might cause early abortion, growth restriction of the fetus or preterm birth (Entrican, 2002). However, injection of HVJ-E vector in the uterine cavity on day 1.5 p.c. (Table I) had no effect on the uterine physiology of implantation, the course of pregnancy or parturition. The vector antigen had already disappeared after 3 days, when the transfected gene was still active, whilst the transfected gene was totally eliminated from the uterus on day 10.5 p.c. These observations suggest that if we use this system to introduce an expression vector that modulates the process of implantation, the gene product is unlikely to persist to interfere with parturition. Moreover, our method does not introduce the exogenous gene into the fetus. These conditions should be ideal for the investigation of implantation physiology. Interestingly, comparison of the tissue distribution between the plasmid DNA and ODN transfected with HVJ-E vector revealed that the patterns appeared to be very different. ß-Galactosidase activity derived from plasmid cDNA was localized in the luminar and glandular epithelium, a condition that would be suitable for the study of the apposition, attachment and early invasion phases of implantation. On the other hand, ODN can be delivered not only to the surface epithelial layer, but also to deeper stromal and myometrial layers, making it suitable for possible application to gene therapy of early endometrial cancer using an antisense ODN or decoy strategy, as the cancer cell layer is relatively thick even in the pre-invasive stage (Gordon and Ireland, 1994). The mechanism leading to this difference is unclear; however, HVJ-E vector appears to become distributed in the myometrial layer (Figure 3b). The amount of gene products from the plasmid might be smaller than the level detectable by ß-galactosidase staining, or cytomegalovirus-promoter activity might be lower in the endometrial stroma and myometrial cells. We introduced HVJ-E vector into the uterine cavity on day 1.5 p.c., when the luminal and glandular epithelium secrete large amounts of anti-adhesive molecules such as MUC-1 (Aplin, 1999). Attachment of HVJ-E appears to occur successfully even in the luminal epithelium (Figure 2b and d), where the embryos will attach. This suggests that this vector could overcome the anti-adhesive activity in the uterine cavity before the opening of the implantation window.
Several strategies are available for in-vivo gene transfer. Retroviruses can deliver transgenes only to proliferating cells, and can lead to long-lasting gene transfer. Adenovirus and adeno-associated virus show higher efficiency, and produce gene expression that is relatively long but transient. Both of these viral systems require relatively complex recombination procedures to introduce the desired cDNA into the vector. Immune reactions against the virus sometimes cause serious problems for the host. On the other hand, naked DNA injection or cationic liposome transfection requires only very simple transgene preparation. Either expression plasmids available for use in mammalian cells or oligodeoxynucleotides (antisense ODN or decoy ODN) can be introduced into the host. These systems induce minimal immune reaction; however, their transfection efficiency is very low (Yla-Herttuala and Martin, 2000). Cationic liposomes have been successfully applied to in-vivo gene transfer: some examples include transfer into the bronchial epithelium and tumour cells (Alton et al., 1993; Hyde et al., 1993; Sugaya et al., 1996; Kikuchi et al., 1999). Hsieh et al. (2002) used GenePorterTM transfection reagent in the uterine cavity and showed that luciferase activity after transfection was 3000 RLU/g protein (3 RLU/mg protein). This level appears to be lower than the level we observed (70 000 RLU/mg protein), although it is impossible to compare the different systems directly. The HVJ-E vector is a novel tool that can easily be used to transfer any kind of expression plasmid, ODN, decoy or protein (Kaneda et al., 2002). One problem with virus-derived vectors may be immunogenicity of the vector in the host, because there are viral proteins on the surface of the vector. Our results show that no local immune response disturbed the course of pregnancy. In the rat, antibodies against HVJ were reported to be detectable 7 days after injection in vivo; however, the titre of the antibody was very low (1000-fold lower than that of a polyclonal antibody produced by rabbits immunized with adjuvant-conjugated HVJ), and cytotoxic T cells against HVJ were not induced (Hirano et al., 1998). These observations might explain the safety of inactivated HVJ in pregnancy, although the effect of HVJ systemic infection on pregnant animals has not been reported. The high fusigenic activity of HVJ-derived vectors has already been shown. Using these vectors [HVJ liposome (Saeki and Kaneda, 1998) and HVJ-E (Kaneda et al., 2002)], exogenous genes were successfully transferred into various organs and animals, as well as human-derived cell lines (Morishita et al., 1997; Mason et al., 1999; Kaneda et al., 2002). Our data reveal that the in-vivo gene transfer system using the HVJ-E vector should become a powerful tool for the study of uterine reproductive physiology and that it has strong promise for the establishment of local gene therapy for infertility and early-stage endometrial cancer.
|
Acknowledgements |
|---|
We thank Satomi Okamoto for assistance with immunohistochemistry. This work was supported in part by Grants-in-Aid for Scientific Research (Nos 14571557 and 15390505) from the Ministry of Education, Science and Culture of Japan (Tokyo, Japan).
|
References |
|---|
|
TopAbstractIntroductionMaterials and methodsResultsDiscussionReferences |
|---|
Alton, E.W., Middleton, P.G., Caplen, N.J., Smith, S.N., Steel, D.M., Munkonge, F.M., Jeffery, P.K., Geddes, D.M., Hart, S.L., Williamson, R. et al. (1993) Non-invasive liposome-mediated gene delivery can correct the ion transport defect in cystic fibrosis. Nature Genet., 5, 135–142.[CrossRef][Web of Science][Medline]
Aplin, J.D. (1999) MUC-1 glycosylation in endometrium: possible roles of the apical glycocalyx at implantation. Hum. Reprod., 14 (Suppl. 2), 17–25.
Bagot, C.N., Troy, P.J. and Taylor, H.S. (2000) Alteration of maternal Hoxa 10 expression by in vivo gene transfection affects implantation. Gene Ther., 7, 1378–1384.[CrossRef][Web of Science][Medline]
Bilinski, P., Roopenian, D. and Gossler, A. (1998) Maternal IL-11R function is required for normal decidua and fetoplacental development in mice. Gene Dev., 12, 2234–2243.
Charnock-Jones, D.S., Sharkey, A.M., Jaggers, D.C., Yoo, H.J., Heap, R.B. and Smith, S.K. (1997) In-vivo gene transfer to the uterine endometrium. Hum. Reprod., 12, 17–20.
Daftary, G.S. and Taylor, H.S. (2001) Efficient liposome-mediated gene transfection and expression in the intact human uterus. Hum. Gene Ther., 12, 2121–2127.[CrossRef][Web of Science][Medline]
Entrican, G. (2002) Immune response during pregnancy and host–pathogen interactions in infectious abortion. J. Comp. Pathol., 126, 79–94.[CrossRef][Web of Science][Medline]
Gordon, M.D. and Ireland, K. (1994) Pathology of hyperplasia and carcinoma of the endometrium. Semin. Oncol., 21, 64–70.[Web of Science][Medline]
Grady, D., Gebretsadik, T., Kerlikowske, K., Ernster, V. and Petitti, D. (1995) Hormone replacement therapy and cancer risk: a meta analysis. Obstet. Gynecol., 85, 304–313.[CrossRef][Web of Science][Medline]
Hirano, T., Fujimoto, J., Ueki, T., Yamamoto, H., Iwasaki, T., Morishita, R., Sawa, Y., Kaneda, Y., Takahashi, H. and Okamoto, E. (1998) Persistent gene expression in rat liver in vivo by repetitive transfections using HVJ-liposome. Gene Ther., 5, 459–464.[CrossRef][Web of Science][Medline]
Hsieh, Y.-Y., Lin, C.-S., Sun, Y.-L., Chang, C.-C., Tsai, H.-D. and Wu, J.C.-H. (2002) In vivo gene transfer of leukemia inhibitory factor (LIF) into mouse endometrium. J. Assist. Reprod. Genet., 19, 79–83.[CrossRef][Web of Science][Medline]
Hyde, S.C., Gill, D.R., Higgins, C.F., Trezise, A.E., MacVinish, L.J., Cuthbert, A.W., Ratcliff, R., Evans, M.J. and Colledge, W.H. (1993) Correction of the ion transport defect in cystic fibrosis transonic mice by gene therapy. Nature, 362, 250–255.[CrossRef][Medline]
Kaneda, Y., Nakajima, T., Nishikawa, T., Yamamoto, S., Ikegami, H., Suzuki, N., Nakamura, H., Morishita, R. and Kotani, H. (2002) Hemagglutinating virus of Japan (HVJ) envelope vector as a versatile gene delivery system. Mol. Ther., 6, 219–226.[CrossRef][Web of Science][Medline]
Kikuchi, A., Aoki, Y., Sugaya, S., Serikawa, T., Takakuwa, K., Tanaka, K., Suzuki, N. and Kikuchi, H. (1999) Development of novel cationic liposomes for efficient gene transfer into peritoneal disseminated tumor. Hum. Gene Ther., 10, 947–955.[CrossRef][Web of Science][Medline]
Lim, H., Paria, B.C., Das, S.K., Dinchuk, J.E., Langenbach, R., Trzaskos, J.M. and Dey, S.K. (1997) Multiple female reproductive failures in cyclooxygenase 2-deficient mice. Cell, 91, 197–208.[CrossRef][Web of Science][Medline]
Lindhart, A., Bentin-Ley, U., Ravn, V., Islin, H., Hviid, T., Rex, S., Bangsboll, S. and Sorensen, S. (2002) Biochemical evaluation of endometrial function at the time of implantation. Fertil. Steril., 78, 221–233.[CrossRef][Web of Science][Medline]
Mason, C.A., Bigras, J.L., O’Blenes, S.B., Zhou, B., Mclntyre, B., Nakamura, N., Kaneda, Y. and Rabinovitch, M. (1999) Gene transfer in utero biologically engineers a patient ductus arteriosus in lambs by arresting fibronectin-dependent neointimal formation. Nature Med., 5, 176–182.[CrossRef][Web of Science][Medline]
Miura, N., Soe, G., Uchida, T. and Okada, Y. (1993) Assessment of membrane fusion efficiency and its use for distinguishing epitopes on the fusion (F) protein of Sendai virus (HVJ). Biochem. Biophys. Res. Commun., 194, 1051–1057.[CrossRef][Web of Science][Medline]
Morishita, R., Sugimoto, T., Aoki, M., Kida, I., Tomita, N., Moriguchi, A., Maeda, K., Sawa, Y., Kaneda, Y., Higaki, J. et al. (1997) In vivo transfection of cis element ‘decoy’ against nuclear factor-
B binding site prevents myocardial infarction. Nature Med., 3, 894–899.[CrossRef][Web of Science][Medline]
Relloso, M. and Esponda, P. (1998) In vivo gene transfer to the mouse oviduct epithelium. Fertil. Steril., 70, 366–368.[CrossRef][Web of Science][Medline]
Saeki, Y. and Kaneda, Y. (1998) Protein-modified liposomes (HVJ-liposomes) for delivery of genes, oligonucleotides and proteins. In Celis, J.E. (ed.), Cell Biology: A Laboratory Handbook. Academic Press, San Diego, Vol. 4, pp. 123–130.
Saeki, Y., Matsumoto, N., Nakano, Y., Mori, M., Awai, K. and Kaneda, Y. (1997) Development and characterization of cationic liposomes conjugated with HVJ (Sendai virus): reciprocal effect of cationic lipid for in vitro and in vivo gene transfer. Hum. Gene Ther., 8, 1965–1972.
Saeki, Y., Wataya-Kaneda, M., Tanaka, K. and Kaneda, Y. (1998) Sustained transgene expression in vitro and in vivo using an Epstein–Barr virus replicon vector system combined with HVJ-liposomes. Gene Ther., 5, 1031–1037.[CrossRef][Web of Science][Medline]
Satokata, I., Benson, G. and Maas, R. (1995) Sexually dimorphic sterility phenotypes in Hoxa-10 deficient mice. Nature, 374, 460–463.[CrossRef][Medline]
Sharkey, A.M. (2000) Uterine gene therapy and implantation. Reprod. Med. Rev., 8, 57–71.[CrossRef]
Stewart, C.L., Kaspar, P., Brunet, L.J., Bhatt, H., Gadi, I., Kontgen, F. and Abbondanzo, S.J. (1992) Blastocyst implantation depends on maternal expression of leukemia inhibitory factor. Nature, 359, 76–79.[CrossRef][Medline]
Sugaya, S., Fujita, K., Kikuchi, A., Ueda, H., Takakuwa, K., Kodama, S. and Tanaka, K. (1996) Inhibition of tumor growth by direct intratumoral gene transfer of herpes simplex virus thymidine kinase gene with DNA–liposome complexes. Hum. Gene Ther., 7, 223–230.[CrossRef][Web of Science][Medline]
Yla-Herttuala, S. and Martin, J.F. (2000) Cardiovascular gene therapy. Lancet, 355, 213–222.[CrossRef][Web of Science][Medline]
Zhu, L.-J., Bagchi, M.K. and Bagchi, I.C. (1998) Attenuation of calcitonin gene expression in pregnant rat uterus leads to a block in embryonic implantation. Endocrinology, 139, 330–339.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  Y. Mitani, J. Maruyama, B. H. Jiang, H. Sawada, H. Shimpo, K. Imanaka-Yoshida, Y. Kaneda, Y. Komada, and K. Maruyama Atrial natriuretic peptide gene transfection with a novel envelope vector system ameliorates pulmonary hypertension in rats J. Thorac. Cardiovasc. Surg., July 1, 2008; 136(1): 142 - 149. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||