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      A patient derived xenograft model of cervical cancer and cervical dysplasia

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          Abstract

          Aim

          To develop a patient derived xenograft (PDX) model of cervical cancer and cervical dysplasia using the subrenal capsule.

          Methods

          Cervical cancer (12 Squamous Cell Carcinoma, 1 Adenocarcinoma, 1 Adenosquamous Carcinoma), 7 cervical dysplasia biopsy and normal cervical tissues were transplanted beneath the renal capsule of immunocompromised NOD/SCID/gamma mice. Resulting tumours were harvested and portions serially transplanted into new recipient mice for up to three in vivo passages. Parent and xenograft tumours were examined by immunohistochemistry for p16 INK41, HPV, and CD-45. Single cell suspensions of mixed mouse and human, or human only cell populations were also transplanted.

          Results

          The overall engraftment rate for the primary cervical cancer PDX model was 71.4 ±12.5% (n = 14). Tumours maintained morphological, histoarchitecture and immunohistochemical features of the parent tumour, and demonstrated invasiveness into local tissues. Single cell suspensions did not produce tumour growth in this model. Mean length of time (32.4 +/- 3.5 weeks) for the transplanted tissue to generate a tumour in the animal was similar between successive transplantations. Three of four xenografted cervical dysplasia tissues generated microscopic cystic structures resembling dysplastic cervical tissue. Normal cervical tissue (4 of 5 xenografted) also developed microscopic cervical tissue grafts.

          Conclusion

          The subrenal capsule can be used for a PDX model of human cervical cancer with a good engraftment rate and the ability to model in vivo characteristics of cervical cancer. For the first time we have demonstrated that cervical dysplasia and normal cervical tissue generated microscopic tissues in a PDX model.

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          Most cited references25

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          Randomised study of radical surgery versus radiotherapy for stage Ib-IIa cervical cancer.

          Stage Ib and IIa cervical carcinoma can be cured by radical surgery or radiotherapy. These two procedures are equally effective, but differ in associated morbidity and type of complications. In this prospective randomised trial of radiotherapy versus surgery, our aim was to assess the 5-year survival and the rate and pattern of complications and recurrences associated with each treatment. Between September, 1986, and December, 1991, 469 women with newly diagnosed stage Ib and IIa cervical carcinoma were referred to our institute. 343 eligible patients were randomised: 172 to surgery and 171 to radical radiotherapy. Adjuvant radiotherapy was delivered after surgery for women with surgical stage pT2b or greater, less than 3 mm of safe cervical stroma, cut-through, or positive nodes. The primary outcome measures were 5-year survival and the rate of complications. The analysis of survival and recurrence was by intention to treat and analysis of complications was by treatment delivered. 170 patients in the surgery group and 167 in the radiotherapy group were included in the intention-to-treat analysis; scheduled treatment was delivered to 169 and 158 women, respectively, 62 of 114 women with cervical diameters of 4 cm or smaller and 46 of 55 with diameters larger than 4 cm received adjuvant therapy. After a median follow-up of 87 (range 57-120) months, 5-year overall and disease-free survival were identical in the surgery and radiotherapy groups (83% and 74%, respectively, for both groups), 86 women developed recurrent disease: 42 (25%) in the surgery group and 44 (26%) in the radiotherapy group. Significant factors for survival in univariate and multivariate analyses were: cervical diameter, positive lymphangiography, and adeno-carcinomatous histotype. 48 (28%) surgery-group patients had severe morbidity compared with 19 (12%) radiotherapy-group patients (p = 0.0004). There is no treatment of choice for early-stage cervical carcinoma in terms of overall or disease-free survival. The combination of surgery and radiotherapy has the worst morbidity, especially urological complications. The optimum therapy for each patient should take account of clinical factors such as menopausal status, age, medical illness, histological type, and cervical diameter to yield the best cure with minimum complications.
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            A primary xenograft model of small-cell lung cancer reveals irreversible changes in gene expression imposed by culture in vitro.

            Traditional approaches to the preclinical investigation of cancer therapies rely on the use of established cell lines maintained in serum-based growth media. This is particularly true of small-cell lung cancer (SCLC), where surgically resected tissue is rarely available. Recent attention has focused on the need for better models that preserve the integrity of cancer stem cell populations, as well as three-dimensional tumor-stromal interactions. Here we describe a primary xenograft model of SCLC in which endobronchial tumor specimens obtained from chemo-naive patients are serially propagated in vivo in immunodeficient mice. In parallel, cell lines grown in conventional tissue culture conditions were derived from each xenograft line, passaged for 6 months, and then reimplanted to generate secondary xenografts. Using the Affymetrix platform, we analyzed gene expression in primary xenograft, xenograft-derived cell line, and secondary xenograft, and compared these data to similar analyses of unrelated primary SCLC samples and laboratory models. When compared with normal lung, primary tumors, xenografts, and cell lines displayed a gene expression signature specific for SCLC. Comparison of gene expression within the xenograft model identified a group of tumor-specific genes expressed in primary SCLC and xenografts that was lost during the transition to tissue culture and that was not regained when the tumors were reestablished as secondary xenografts. Such changes in gene expression may be a common feature of many cancer cell culture systems, with functional implications for the use of such models for preclinical drug development.
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              Carcinoma of the cervix uteri. FIGO 26th Annual Report on the Results of Treatment in Gynecological Cancer.

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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: Writing – original draftRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: Methodology
                Role: ConceptualizationRole: Supervision
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: MethodologyRole: Project administrationRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                26 October 2018
                2018
                : 13
                : 10
                : e0206539
                Affiliations
                [1 ] The Ritchie Centre, Hudson Institute for Medical Research, Clayton, Victoria, Australia
                [2 ] Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria, Australia
                [3 ] Melbourne Pathology, Collingwood, Australia
                Istituto Nazionale Tumori IRCCS Fondazione Pascale, ITALY
                Author notes

                Competing Interests: J.A.T. is employed by Melbourne Pathology. Work on this project was performed on her own time, and the only support provided was salary. This does not alter our adherence to all PLOS ONE policies on data sharing and materials. We confirm that there are no other competing interests to declare.

                Author information
                http://orcid.org/0000-0002-7146-5978
                http://orcid.org/0000-0002-3590-2077
                Article
                PONE-D-18-18796
                10.1371/journal.pone.0206539
                6203389
                30365542
                645896c8-50f1-4aac-b606-8644be555ab9
                © 2018 Larmour et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 2 July 2018
                : 15 October 2018
                Page count
                Figures: 4, Tables: 3, Pages: 16
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100000925, National Health and Medical Research Council;
                Award ID: 1042298
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001104, RANZCOG Research Foundation;
                Award ID: Mary Elizabeth Courier Scholarship
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100000991, Faculty of Medicine, Nursing and Health Sciences, Monash University;
                Award ID: David Healy Memorial Scholarship
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100004752, State Government of Victoria;
                Award ID: Operational Infrastructure Support Program
                This work was supported by National Health and Medical Research Council Senior Research Fellowship (1042298) (C.E.G.), Royal Australian and New Zealand College of Obstetricians and Gynaecologists Research Foundation Mary Elizabeth Courier Scholarship (L.I.L.), Monash University David Healy Memorial Scholarship (L.I.L) and the Victorian Government's Operational Infrastructure Support Program. Melbourne Pathology provided support in the form of salary for author J.A.T., but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of the authors are articulated in the ‘author contributions’ section. The work performed by J.A.T was on her own time. The funding bodies had no role in any aspect of the study.
                Categories
                Research Article
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Carcinomas
                Squamous Cell Carcinomas
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Gynecological Tumors
                Cervical Cancer
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                Diagnostic Medicine
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