Abstract
Uterine fibroids are the most prevalent benign tumors in women of reproductive age. The current knowledge on the fibroid disease mechanism has derived from studies of the Eker rat model where a unique germ line defect in the tuberous sclerosis 2 (Tsc2) tumor suppressor gene leads to the development of leiomyosarcoma, leiomyoma, and renal cancer. To study fibroids of human origin, we sought to establish fibroid xenografts in immune-compromised mice. We determined that lentiviral-mediated transduction of a green fluorescence protein (GFP)-luciferase (LUC) fusion gene and bioluminescence-based whole animal imaging allowed for the monitoring of transplanted fibroid cells in mice. We used this in vivo imaging approach to test a series of transplantation protocols and found that only freshly dissociated fibroid cells, but not the fibroid-derived smooth muscle cells grown in ex vivo cultures, can generate stable xenografts in subcutaneous Matrigel implants. Formation of the fibroid-xenografts requires the implantation of 17β-estradiol-releasing pellets in the recipient mice. Furthermore, freshly dissociated myometrial cells do not form xenografts under the experimental conditions. The xenograft protocol developed from this study provides an avenue for investigating the pathogenesis and drug responses of human fibroids.
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Cramer SF, Patel A. The nonrandom regional distribution of uterine leiomyomas: a clue to histogenesis? Hum Pathol. 1992;23(6): 635–638.
Olive DL. Analysis of clinical fertility trials: a methodologic review. Fertil Steril. 1986;45(2):157–171.
Morales AJ, Kettel LM. Quality of life assessment. Semin Reprod Endocrinol. 1996;14(2):155–159.
Dembek CJ, Pelletier EM, Isaacson KB, Spies JB. Payer costs in patients undergoing uterine artery embolization, hysterectomy, or myomectomy for treatment of uterine fibroids. J Vasc Interv Radiol. 2007;18(10):1207–1213.
Hartmann KE, Birnbaum H, Ben-Hamadi R, et al. Annual costs associated with diagnosis of uterine leiomyomata. Obstet Gynecol. 2006;108(4):930–937.
Linder D, Gartler SM. Glucose-6-phosphate dehydrogenase mosaicism: utilization as a cell marker in the study of leiomyomas. Science. 1965;150(692):67–69.
Wilson EA, Yang F, Rees ED. Estradiol and progesterone binding in uterine leiomyomata and in normal uterine tissues. Obstet Gynecol. 1980;55(1):20–24.
Rein MS, Barbieri RL, Friedman AJ. Progesterone: a critical role in the pathogenesis of uterine myomas. Am J Obstet Gynecol. 1995;172(1 pt 1):14–18.
Andersen J, DyReyes VM, Barbieri RL, Coachman DM, Miksicek RJ. Leiomyoma primary cultures have elevated transcriptional response to estrogen compared with autologous myometrial cultures. J Soc Gynecol Investig. 1995;2(3): 542–551.
Ichimura T, Kawamura N, Ito F, et al. Correlation between the growth of uterine leiomyomata and estrogen and progesterone receptor content in needle biopsy specimens. Fertil Steril. 1998;70(5):967–971.
Vu K, Greenspan DL, Wu TC, Zacur HA, Kurman RJ. Cellular proliferation, estrogen receptor, progesterone receptor, and bcl-2 expression in GnRH agonist-treated uterine leiomyomas. Hum Pathol. 1998;29(4):359–363.
Mashal RD, Fejzo ML, Friedman AJ, et al. Analysis of androgen receptor DNA reveals the independent clonal origins of uterine leiomyomata and the secondary nature of cytogenetic aberrations in the development of leiomyomata. Genes Chromosomes Cancer. 1994;11(1):1–6.
Wang T, Zhang X, Obijuru L, et al. A micro-RNA signature associated with race, tumor size, and target gene activity in human uterine leiomyomas. Genes Chromosomes Cancer. 2007;46(4):336–347.
Vanharanta S, Pollard PJ, Lehtonen HJ, et al. Distinct expression profile in fumarate-hydratase-deficient uterine fibroids. Hum Mol Genet. 2006;15(1):97–103.
Ingraham SE, Lynch RA, Surti U, et al. Identification and characterization of novel human transcripts embedded within HMGA2 in t(12;14)(ql5;q24.1) uterine leiomyoma. Mutat Res. 2006;602(1–2):43–53.
Everitt JI, Wolf DC, Howe SR, Goldsworthy TL, Walker C. Rodent model of reproductive tract leiomyomata. Clinical and pathological features. Am J Pathol. 1995;146(6): 1556–1567.
Yeung RS, Xiao GH, Jin F, Lee WC, Testa JR, Knudson AG. Predisposition to renal carcinoma in the Eker rat is determined by germ-line mutation of the tuberous sclerosis 2 (TSC2) gene. Proc Natl Acad Sci U S A. 1994;91(24):11413–11416.
Howe SR, Gottardis MM, Everitt JI, Goldsworthy TL, Wolf DC, Walker C. Rodent model of reproductive tract leiomyomata. Establishment and characterization of tumor-derived cell lines. Am J Pathol. 1995;146(6):1568–1579.
Wei J, Chiriboga L, Mizuguchi M, Yee H, Mittal K. Expression profile of tuberin and some potential tumorigenic factors in 60 patients with uterine leiomyomata. Mod Pathol. 2005;18(2):179–188.
Hassan MH, Eyzaguirre E, Arafa HM, Hamada FM, Salama SA, Al-Hendy A. Memy I: a novel murine model for uterine leiomyoma using adenovirus-enhanced human fibroid explants in severe combined immune deficiency mice. Am J Obstet Gynecol. 2008;199(2):156 el-e8.
Legrand N, Weijer K, Spits H. Experimental model for the study of the human immune system: production and monitoring of “human immune system” Rag2-/-gamma c-/- mice. Methods Mol Biol. 2008;415:65–82.
Rice BW, Cable MD, Nelson MB. In vivo imaging of light-emitting probes. J Biomed Opt. 2001;6(4):432–440.
Kuo C, Coquoz O, Troy TL, Xu H, Rice BW. Three-dimensional reconstruction of in vivo bioluminescent sources based on multispectral imaging. J Biomed Opt. 2007;12(2):024007.
Shinkai Y, Rathbun G, Lam KP, et al. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell. 1992;68(5):855–867.
Soudais C, Shiho T, Sharara LI, et al. Stable and functional lymphoid reconstitution of common cytokine receptor gamma chain deficient mice by retroviral-mediated gene transfer. Blood. 2000;95(10):3071–3077.
Baenziger S, Tussiwand R, Schlaepfer E, et al. Disseminated and sustained HIV infection in CD34+ cord blood cell-transplanted Rag2-/-gamma c-/- mice. Proc Natl Acad Sci U S A. 2006;103(43):15951–15956.
Traggiai E, Chicha L, Mazzucchelli L, et al. Development of a human adaptive immune system in cord blood cell-transplanted mice. Science. 2004;304(5667):104–107.
D’Hallewin MA, El Khatib S, Leroux A, Bezdetnaya L, Guillemin F. Endoscopic confocal fluorescence microscopy of normal and tumor bearing rat bladder. J Urol. 2005; 174(2):736–740.
Tanaka M, Gee JR, De La Cerda J, et al. Noninvasive detection of bladder cancer in an orthotopic murine model with green fluorescence protein cytology. J Urol. 2003;170(3): 975–978.
Bukowski EJ, Bright FV. Minimizing urine autofluorescence under multi-photon excitation conditions. Appl Spectrosc. 2004;58(9):1101–1105.
Guamaccia MM, Rein MS. Traditional surgical approaches to uterine fibroids: abdominal myomectomy and hysterectomy. Clin Obstet Gynecol. 2001;44(2):385–400.
Yeh J, Rein M, Nowak R. Presence of messenger ribonucleic acid for epidermal growth factor (EGF) and EGF receptor demonstrable in monolayer cell cultures of myometria and leiomyomata. Fertil Steril. 1991;56(5):997–1000.
Giudice LC, Irwin JC, Dsupin BA, et al. Insulin-like growth factor (IGF), IGF binding protein (IGFBP), and IGF receptor gene expression and IGFBP synthesis in human uterine leiomyomata. Hum Reprod. 1993;8(11):1796–1806.
Lethaby AE, Vollenhoven BJ. An evidence-based approach to hormonal therapies for premenopausal women with fibroids. Best Pract Res Clin Obstet Gynaecol. 2008;22(2):307–331.
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Suo, G., Sadarangani, A., LaMarca, B. et al. Murine Xenograft Model for Human Uterine Fibroids: An In Vivo Imaging Approach. Reprod. Sci. 16, 827–842 (2009). https://doi.org/10.1177/1933719109336615
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DOI: https://doi.org/10.1177/1933719109336615