Name: handling of the cotton rat in studies for the pre-clinical evaluation of oncolytic viruses
Uploaded: 2014-11-24T15:00:00.000+00:00
Duration: 6 min 13 s
Description: Watch this Scientific Journal Video about Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses at JoVE.com

Breanne Cuddington holds a fellowship from the Canadian Breast Cancer Foundation. This work was sponsored by operating grants from the Cancer Research Society and the Canadian Cancer Society Research Institute (formerly the Canadian Breast Cancer Research Alliance). We thank Ann Tollefson (Saint Louis University School of Medicine) for LCRT cells and Dr. Kathleen Delaney and Marion Corrick for technical assistance with cotton rat housing and sedation.

<ol>
	<li>Cervantes-Garcia, D., Ortiz-Lopez, R., Mayek-Perez, N., Rojas-Martinez, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oncolytic+virotherapy.">Oncolytic virotherapy.</a> Ann Hepatol. 7 (1), 34-45 (2008).</li><li>Vaha-Koskela, M. J., Heikkila, J. E., Hinkkanen, A. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oncolytic+viruses+in+cancer+therapy.">Oncolytic viruses in cancer therapy.</a> Cancer Lett. 254 (2), 178-216 (2007).</li><li>Abril, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Both+viral+and+host+factors+contribute+to+neurovirulence+of+bovine+herpesviruses+1+and+5+in+interferon+receptor-deficient+mice.">Both viral and host factors contribute to neurovirulence of bovine herpesviruses 1 and 5 in interferon receptor-deficient mice.</a> J Virol. 78 (7), 3644-3653 (2004).</li><li>Nakamichi, K., Matsumoto, Y., Otsuka, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defective+infection+of+bovine+herpesvirus+1+in+non-permissive+murine+cells.">Defective infection of bovine herpesvirus 1 in non-permissive murine cells.</a> J Vet Med Sci. 63 (10), 1139-1142 (2001).</li><li>Boukhvalova, M. S., Blanco, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cotton+rat+sigmodon+hispidus+model+of+respiratory+syncytial+virus+infection.">The cotton rat sigmodon hispidus model of respiratory syncytial virus infection.</a> Curr Top Microbiol Immunol. 372, 347-358 (2013).</li><li>Papp, Z., Babiuk, L. A., Baca-Estrada, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+immunity+in+the+respiratory+tract+and+protection+from+bovine+herpesvirus+type+1+infection+by+different+routes+of+immunization+with+recombinant+adenovirus.">Induction of immunity in the respiratory tract and protection from bovine herpesvirus type 1 infection by different routes of immunization with recombinant adenovirus.</a> Viral Immunol. 11 (2), 79-91 (1998).</li><li>Hughes, T. C. R., Lilley, C. E., Ponce, R., Kaufman, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+analysis+of+an+oncolytic+herpesvirus+encoding+granulocyte-macrophage+colony+stimulating+factor+for+the+treatment+of+malignant+melanoma.">Critical analysis of an oncolytic herpesvirus encoding granulocyte-macrophage colony stimulating factor for the treatment of malignant melanoma.</a> Journal of Oncolytic Virotherapy. 3, 11-20 (2014).</li><li>Jones, C., Chowdhury, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+the+biology+of+bovine+herpesvirus+type+1+(BHV-1),+its+role+as+a+cofactor+in+the+bovine+respiratory+disease+complex+and+development+of+improved+vaccines.">A review of the biology of bovine herpesvirus type 1 (BHV-1), its role as a cofactor in the bovine respiratory disease complex and development of improved vaccines.</a> Anim Health Res Rev. 8 (2), 187-205 (2007).</li><li>Jones, C., Chowdhury, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bovine+herpesvirus+type+1+(BHV-1)+is+an+important+cofactor+in+the+bovine+respiratory+disease+complex.">Bovine herpesvirus type 1 (BHV-1) is an important cofactor in the bovine respiratory disease complex.</a> Vet Clin North Am Food Anim Pract. 26 (2), 303-321 (2010).</li><li>Hushur, O., Takashima, Y., Matsumoto, Y., Otsuka, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restriction+of+bovine+herpesvirus+1+(BHV-1)+growth+in+non-permissive+cells+beyond+the+expression+of+immediate+early+genes.">Restriction of bovine herpesvirus 1 (BHV-1) growth in non-permissive cells beyond the expression of immediate early genes.</a> J Vet Med Sci. 66 (4), 453-455 (2004).</li><li>Cuddington, B. P., Dyer, A. L., Workenhe, S. T., Mossman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oncolytic+bovine+herpesvirus+type+1+infects+and+kills+breast+tumor+cells+and+breast+cancer-initiating+cells+irrespective+of+tumor+subtype.">Oncolytic bovine herpesvirus type 1 infects and kills breast tumor cells and breast cancer-initiating cells irrespective of tumor subtype.</a> Cancer Gene Ther. 20 (5), 282-289 (2013).</li><li>Cuddington, B. P., Mossman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Permissiveness+of+Human+Cancer+Cells+to+Oncolytic+Bovine+Herpesvirus+1+Is+Mediated+in+Part+by+KRAS+Activity.">Permissiveness of Human Cancer Cells to Oncolytic Bovine Herpesvirus 1 Is Mediated in Part by KRAS Activity.</a> J Virol. 88 (12), 6885-6895 (2014).</li><li>Small, E. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+phase+I+trial+of+intravenous+CG7870,+a+replication-selective,+prostate-specific+antigen-targeted+oncolytic+adenovirus,+for+the+treatment+of+hormone-refractory,+metastatic+prostate+cancer.">A phase I trial of intravenous CG7870, a replication-selective, prostate-specific antigen-targeted oncolytic adenovirus, for the treatment of hormone-refractory, metastatic prostate cancer.</a> Mol Ther. 14 (1), 107-117 (2006).</li><li>Freytag, S. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phase+I+study+of+replication-competent+adenovirus-mediated+double+suicide+gene+therapy+for+the+treatment+of+locally+recurrent+prostate+cancer.">Phase I study of replication-competent adenovirus-mediated double suicide gene therapy for the treatment of locally recurrent prostate cancer.</a> Cancer Res. 62 (17), 4968-4976 (2002).</li><li>Benjamin, R., Helman, L., Meyers, P., Reaman, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+phase+I/II+dose+escalation+and+activity+study+of+intravenous+injections+of+OCaP1+for+subjects+with+refractory+osteosarcoma+metastatic+to+lung.">A phase I/II dose escalation and activity study of intravenous injections of OCaP1 for subjects with refractory osteosarcoma metastatic to lung.</a> Hum Gene Ther. 12 (12), 1591-1593 (2001).</li><li>Prince, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Cotton+Rat+in+Biomedical+Research.">The Cotton Rat in Biomedical Research.</a> Animal Welfare Information Center Newsletter. 5 (2), 3-5 (1994).</li><li>Tsai, J. C., Garlinghouse, G., McDonnell, P. J., Trousdale, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+experimental+animal+model+of+adenovirus-induced+ocular+disease.+The+cotton+rat.">An experimental animal model of adenovirus-induced ocular disease. The cotton rat.</a> Arch Ophthalmol. 110 (8), 1167-1170 (1992).</li><li>Ginsberg, H. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+model+for+investigating+the+molecular+pathogenesis+of+adenovirus+pneumonia.">A mouse model for investigating the molecular pathogenesis of adenovirus pneumonia.</a> Proc Natl Acad Sci U S A. 88 (5), 1651-1655 (1991).</li><li>Russell, S. J., Peng, K. W., Bell, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oncolytic+virotherapy.">Oncolytic virotherapy.</a> Nat Biotechnol. 30 (7), 658-670 (2012).</li><li>Mossman, K. L., Saffran, H. A., Smiley, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herpes+simplex+virus+ICP0+mutants+are+hypersensitive+to+interferon.">Herpes simplex virus ICP0 mutants are hypersensitive to interferon.</a> J Virol. 74 (4), 2052-2056 (2000).</li><li>Mossman, K. L., Smiley, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Herpes+simplex+virus+ICP0+and+ICP34.5+counteract+distinct+interferon-induced+barriers+to+virus+replication.">Herpes simplex virus ICP0 and ICP34.5 counteract distinct interferon-induced barriers to virus replication.</a> J Virol. 76 (4), 1995-1998 (2002).</li><li>Hummel, J. L., Safroneeva, E., Mossman, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+ICP0-Null+HSV-1+and+interferon+signaling+defects+in+the+effective+treatment+of+breast+adenocarcinoma.">The role of ICP0-Null HSV-1 and interferon signaling defects in the effective treatment of breast adenocarcinoma.</a> Mol Ther. 12 (6), 1101-1110 (2005).</li><li>Papp, Z., Middleton, D. M., Mittal, S. K., Babiuk, L. A., Baca-Estrada, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mucosal+immunization+with+recombinant+adenoviruses:+induction+of+immunity+and+protection+of+cotton+rats+against+respiratory+bovine+herpesvirus+type+1+infection.">Mucosal immunization with recombinant adenoviruses: induction of immunity and protection of cotton rats against respiratory bovine herpesvirus type 1 infection.</a> J Gen Virol. 78 (11), 2933-2943 (1997).</li><li>Papp, Z., Babiuk, L. A., Baca-Estrada, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+pre-existing+adenovirus-specific+immunity+on+immune+responses+induced+by+recombinant+adenovirus+expressing+glycoprotein+D+of+bovine+herpesvirus+type+1.">The effect of pre-existing adenovirus-specific immunity on immune responses induced by recombinant adenovirus expressing glycoprotein D of bovine herpesvirus type 1.</a> Vaccine. 17 (7-8), 933-943 (1999).</li><li>Mittal, S. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+systemic+and+mucosal+immune+responses+in+cotton+rats+immunized+with+human+adenovirus+type+5+recombinants+expressing+the+full+and+truncated+forms+of+bovine+herpesvirus+type+1+glycoprotein+gD.">Induction of systemic and mucosal immune responses in cotton rats immunized with human adenovirus type 5 recombinants expressing the full and truncated forms of bovine herpesvirus type 1 glycoprotein gD.</a> Virology. 222 (2), 299-309 (1996).</li><li>Steel, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Syngeneic+Cotton+Rat+Cancer+Model+for+Replicating+Adenoviral+Vectors.">Syngeneic Cotton Rat Cancer Model for Replicating Adenoviral Vectors.</a> Molecular Therapy. 13 (1), 123 (2006).</li><li>Toth, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cotton+rat+tumor+model+for+the+evaluation+of+oncolytic+adenoviruses.">Cotton rat tumor model for the evaluation of oncolytic adenoviruses.</a> Hum Gene Ther. 16 (1), 139-146 (2005).</li><li>Toth, K., Spencer, J. F., Wold, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunocompetent,+semi-permissive+cotton+rat+tumor+model+for+the+evaluation+of+oncolytic+adenoviruses.">Immunocompetent, semi-permissive cotton rat tumor model for the evaluation of oncolytic adenoviruses.</a> Methods Mol Med. 130, 157-168 (2007).</li><li>Steel, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunocompetent+syngeneic+cotton+rat+tumor+models+for+the+assessment+of+replication-competent+oncolytic+adenovirus.">Immunocompetent syngeneic cotton rat tumor models for the assessment of replication-competent oncolytic adenovirus.</a> Virology. 369 (1), 131-142 (2007).</li><li>Workenhe, S. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunogenic+HSV-mediated+oncolysis+shapes+the+antitumor+immune+response+and+contributes+to+therapeutic+efficacy.">Immunogenic HSV-mediated oncolysis shapes the antitumor immune response and contributes to therapeutic efficacy.</a> Mol Ther. 22 (1), 123-131 (2014).</li><li>Sobol, P. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptive+antiviral+immunity+is+a+determinant+of+the+therapeutic+success+of+oncolytic+virotherapy.">Adaptive antiviral immunity is a determinant of the therapeutic success of oncolytic virotherapy.</a> Mol Ther. 19 (2), 335-344 (2011).</li><li>Prince, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Cotton+Rat+in+Biomedical+Research.">The Cotton Rat in Biomedical Research.</a> Animal Welfare Information Center Newsletter. 5 (2), (1994).</li></ol>

The authors acknowledge there are no financial conflicts of interest related to this research.

Cotton rats are highly excitable and have a strong flight response. Therefore, special care should be taken to minimize any undue stress on the animal. The cage setup described will allow for safe and easy capture of the animals, with the placement of the enrichment tube being of the utmost importance. When setting up cages, ensure that the enrichment tubes meet the size and shape requirements, and are placed in proper orientation in the cage. It is also important to ensure that any technicians who might be aiding in ani...

الفيروسات حال الورم (OV) تكرار انتقائي في الخلايا السرطانية من خلال استغلال الاختلافات البيوكيميائية بين الخلايا الطبيعية والسرطانية 1،2. هناك نوعان من OVS: تلك التي لا تحتاج إلى طفرة لتحقيق انحلال الورم انتقائية، ويشار إلى الفيروسات التي تحدث بشكل طبيعي من النوع البري وتلك التي يجب أن هندستها لتحقيق انحلال الورم انتقائي. جمع الطفرات ضمن نوع الورم نظرا يحدد طبيعة ميزة النمو انتقائية على الخلايا الطبيعية لOV 2. وقد تجلى سلامة والاستفادة من استخدام OVS في التجارب السريرية 3-7. وعلى الرغم من التقدم في مجال علاج بالفيروسات حال الورم توجد الفجوات بين النتائج ما قبل السريرية والسريرية، مما يوحي بأن هناك حاجة إلى نماذج أفضل لتقييم فعالية مضادة للورم من OVS. البقري نوع هربس 1 (BHV-1) هو عضو في عائلة فيروسات هربسية، وAlphaherpesviridae فصيلة. BHV-1 initiآتش البقري مجمع أمراض الجهاز التنفسي في الماشية، ويظهر في مجموعة متنوعة واسعة من أعراض تشبه نزلة برد 8،9. BHV-1 يربط مرفق ودخول مستقبلات يستخدمها HSV-1، مثل كبريتات heparan وnectin-1 10. ومع ذلك، فإنه يربط CD155 في مكان nectin-2 10. BHV-1 لديها مجموعة والمضيف ضيق جدا بحيث أنه غير قادر على الدخول بكفاءة وبدء النسخ المتماثل في الخلايا الطبيعية وتحويلها الفئران 3،4،10. وهذا يجعل من استخدام نماذج الفئران التقليدية إشكالية. وقد تجلى القدرة حال الورم من BHV-1 في المختبر 11،12. وقد تبين BHV-1 لبدء النسخ المتماثل في وقتل الخلايا السرطانية البشرية من مجموعة متنوعة من أصول النسيجية، بما في ذلك خلايا سرطان الثدي وسرطان الثدي الشروع في الخلايا 11،12. ومع ذلك، يجب تقييم القدرة انتيتومور من BHV-1 في الجسم الحي في سياق مضيف مناعيا. اتش الإنسان (الإعلان)، والتيهناك 57 الأنماط المصلية التي تم تحديدها، تتسبب الأكثر شيوعا أمراض الجهاز التنفسي لدى البشر. تم تقييم ناقلات الإعلان حال الورم لفعالية مضادة للورم مع العديد التقدم في التجارب السريرية 13-15. وعلى الرغم من بيانات ما قبل السريرية واعدة، وتراجعت النتائج السريرية قصيرة من التوقعات. وعادة ما يتم استخدام نماذج طعم أجنبي الورم البشري لدراسة فعالية مضادة للورم من ناقلات الإعلان، على الرغم من أنها تظهر الموهن الاستجابات المناعية للفيروس 16،17. وعلاوة على ذلك، ونماذج الفئران مسانج غير قابلة للمتساهلة للإصابة الإعلان، مما يجعل تقييم المضيف الاستجابات المناعية باستخدام هذه النماذج غير عملي 17،18. تم التعرف على الجهاز المناعي المضيف كآلية الأكثر تأثيرا التي OVS يثير ورم الخلايا الموت 19. ردود انتيتومور بين tolerized وغير tolerized مستضد المرتبطة الورم (TAA) نماذج تختلف ويمكن أن تؤثر بشكل كبير على نجاح العلاج OV. وHSV-1 OV KM100 (ICP0 n212VP16 في عام 1814 20) 20،21 أثار الورم الانحدار في 80٪ من الفئران الحاملة للورم في الفئران التورام الأوسط T مستضد نموذج سرطان الثدي 22. ومع ذلك، في HER-2 نماذج / NEU، وفعالية مضادة للورم من KM100 تنوعت بين الانحدار الكامل 20٪ في الفئران مسانج وورم في ركود المعدلة وراثيا والفئران HER2 tolerized. معا هذه البيانات تبرز أهمية تقييم كامل OVS باستخدام نماذج حيوانية أن ألخص أفضل المشهد المناعة البشري أن نفهم تماما ما هي الميزات تحديد النجاح العلاجي. يتم استخدام الفئران القطن (Sigmodon hispidus)، السكان الأصليين لأمريكا الشمالية والجنوبية، والأكثر شيوعا كنموذج للعدوى فيروس الجهاز التنفسي المخلوي (كما استعرض في 5). وتستخدم الفئران القطن أيضا في مجال البحوث لمكافحة BHV-1 التطعيم لأنها تلخيص علم الأمراض المرتبطة بأمراض الجهاز التنفسي البقري معقدة 6،23. وعلاوة على ذلك، BHV-1 إصابة الفئران القطنيولد مناعة، الأمر الذي أدى المخاطية المستدام والاستجابات المناعية الجهازية 6،23-25. وقد استمدت خطوط الخلايا من يفية عفوية وosteosarcomas من الغدة الثديية (LCRT) والعظام (CCRT وVCRT)، على التوالي 26. وقد استخدمت الفئران القطن لتقييم فعالية في الجسم الحي من ناقلات الإعلان حال الورم كما هي عرضة للإصابة الإعلان ويحمل علم أمراض مماثلة على البشر 27-29. استخدام نماذج المناعة لتقييم ما قبل السريرية من OVS ليسوا سوى أقل يدل على الردود السريرية للعلاج لكنها لا تأخذ في الاعتبار دور الجهاز المناعي في علاج بالفيروسات حال الورم 30،31. ولذلك، فإن مسانج ونماذج الفئران القطن tolerized ورم سرطان الثدي وعظمية والنماذج ذات الصلة التي لتقييم فعالية ما قبل السريرية من OVS، مثل BHV-1 والإعلان التي لا يمكن دراستها باستخدام نماذج الفئران التقليدية.

<table><tbody><tr><td>Dulbecco&amp;rsquo;s modified Eagle&amp;rsquo;s medium&amp;nbsp;</td><td>Gibco</td><td>11965-092</td><td>May use any brand&amp;nbsp;</td></tr><tr><td>1X Phosphate Buffered Saline&amp;nbsp;</td><td></td><td></td><td>Can prepare in lab, filter to sterilize</td></tr><tr><td>200 mM L-glutamine</td><td>Gibco</td><td>25030164</td><td>May use any brand</td></tr><tr><td>100x Antibiotic-Antimycotic&amp;nbsp;</td><td>Gibco</td><td>15240-062</td><td>May use any brand</td></tr><tr><td>Fetal bovine serum</td><td>Quality Biological Inc.</td><td>110-001-101HI</td><td>May use any brand</td></tr><tr><td>T-150cm&lt;sup&gt;2&lt;/sup&gt; tissue culture flask</td><td>Fisher Scientific</td><td>14-826-80</td><td>May use any brand</td></tr><tr><td>1X TypLE Express</td><td>Life Technologies</td><td>12604-013</td><td></td></tr><tr><td>12-well cell culture plate, flat bottom</td><td>Fisher Scientific</td><td>08-772-29</td><td>May use any brand, must be tissue culture treated</td></tr><tr><td>alamarBlue</td><td>Life Technologies</td><td>DAL1025</td><td>May use an alternative reagent for determination of cell viability</td></tr><tr><td>8640 Teklad 22/5 Rodent diet</td><td>Harlan&amp;nbsp;</td><td>8640</td><td></td></tr><tr><td>1/8&amp;rdquo; corncob rodent bedding</td><td>Harlan</td><td>7092</td><td></td></tr><tr><td>Nestlets</td><td>Ancare</td><td>-</td><td>Made of pulped virgin cotton fiber, dust-free and autoclavable</td></tr><tr><td>50 mL Conical tubes</td><td>Fisher Scientific</td><td>14-432-22</td><td>May use any brand, must be sterile</td></tr><tr><td>Isoflurane USP, 99.9 %, inhalation anesthetic</td><td>Pharmaceutical Partners of Canada Inc.</td><td>M60302</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td>Can prepare in lab</td></tr><tr><td>10 % Neutral Buffered Formalin</td><td>Sigma-Aldrich</td><td>HT501128</td><td>May use any brand</td></tr><tr><td>NAPCO NapFlow 1200 Class II A/B3 Biosafety Microbiological Safety Cabinet (cell culture hood)</td><td>NAPCO</td><td>Model used not currently available</td><td>May use any brand</td></tr><tr><td>Thermo Fisher Scientific Precision Heated Water Bath</td><td>Fisher Scientific</td><td>Model used not currently available&amp;nbsp;</td><td>May use any brand</td></tr><tr><td colspan="4">[header]</td></tr><tr><td>Reichert Bright-line Hemacytometer</td><td>Sigma-Aldrich</td><td>Z359629</td><td>May use any brand</td></tr><tr><td>Typhoon Trio BioAnalyzer&amp;nbsp;</td><td>GE Healthcare Life Sciences</td><td>Model used not currently available&amp;nbsp;</td><td>May use any fluorescence plate reader</td></tr><tr><td>Tecan Safire&lt;sup&gt;2&lt;/sup&gt; Multi-detection Microplate Reader</td><td>Tecan</td><td>Model used not currently available&amp;nbsp;</td><td>May use any fluorescence plate reader</td></tr><tr><td>Allegra 6R benchtop centrifuge</td><td>Beckman Coulter</td><td>366816</td><td>May use any brand</td></tr><tr><td>Table Top Anaesthesia machine</td><td>VetEquip</td><td>Model used not currently available&amp;nbsp;</td><td>May use any brand, must be portable</td></tr><tr><td>Wahl Peanut Mini Clippers</td><td>Wahl</td><td></td><td>May use any brand of small clippers</td></tr><tr><td>Insulin syringes 29 G x 1/2&#039;, 0.3 mL</td><td>BD</td><td>329464</td><td>May use any brand. Insulin syringes are recommended as they make injections easier through the rat&amp;rsquo;s tough skin.&amp;nbsp;</td></tr><tr><td>Cotton swabs</td><td>MedPro</td><td>018-425</td><td>May use any brand</td></tr><tr><td>Sharp-Pointed Dissecting Scissors</td><td>Fisher Scientific</td><td>8940</td><td>May use any brand</td></tr><tr><td>Dissecting Tissue Forceps</td><td>Fisher Scientific</td><td>13-812-41</td><td>May use any brand</td></tr></tbody></table>

handling of the cotton rat in studies for the pre-clinical evaluation of oncolytic viruses

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact. For this strategy to be successful, recent studies have shown that involvement of the host immune system is essential. Therefore, oncolytic virotherapy should be evaluated within the context of an immunocompetent model. Furthermore, the study of antitumor therapies in tolerized animal models may better recapitulate results seen in clinical trials. Cotton rats, commonly used to study respiratory viruses, are an attractive model to study oncolytic virotherapy as syngeneic models of mammary carcinoma and osteosarcoma are well established. However, there is a lack of published information on the proper handling procedure for these highly excitable rodents. The handling and capture approach outlined minimizes animal stress to facilitate experimentation. This technique hinges upon the ability of the researcher to keep calm during handling and perform procedures in a timely fashion. Finally, we describe how to prepare cotton rat mammary tumor cells for consistent subcutaneous tumor formation, and how to perform intratumoral and intraperitoneal injections. These methods can be applied to a wide range of studies furthering the development of the cotton rat as a relevant pre-clinical model to study antitumor therapy.

ونظرا لطبيعة منفعل للغاية من الفئران القطن، ويجري على دراية والاستفادة من إجراءات الأمثل للحد من وطأة هذه الحيوانات سوف سهولة في استخدامها كنموذج حيوان ما قبل السريرية. واستخدام التقنيات السليمة للتعامل معها تقليل خطر أيضا للباحث. <p class="jove_content" style=";text-align:right;direction...

Watch this Scientific Journal Video about Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses at JoVE.com

Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses

Cotton rats are extremely excitable and have a strong flight-or-fight response. A handling method optimized to reduce the stress of the animals is described which will make cotton rats more accessible as a preclinical model.

التعامل مع الفأر القطن في الدراسات لتقييم ما قبل السريرية من حال الورم الفيروسات

Oncolytic viruses are a novel anticancer therapy with the ability to target tumor cells, while leaving healthy cells intact.  For this strategy to be ...

handling-cotton-rat-studies-for-pre-clinical-evaluation-oncolytic

Research

JoVE Journal

Medicine

9.3K Views.  McMaster University. Cotton rats are extremely excitable and have a strong flight-or-fight response. A handling method optimized to reduce the stress of the animals is described which will make cotton rats more accessible as a preclinical model. 

Video: Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses

Genome-wide RNAi Screening to Identify Host Factors That Modulate Oncolytic Virus Therapy

High-throughput genome-wide RNAi (RNA interference) screening technology has been widely used for discovering host factors that impact&#160;virus replication. Here we present the application of this technology to uncovering host targets that specifically modulate the replication of Maraba virus, an oncolytic rhabdovirus, and vaccinia virus with the goal of enhancing therapy. While the protocol has been tested for use with oncolytic Maraba virus and oncolytic vaccinia virus, this approach is applicable to other oncolytic viruses and can also be utilized for identifying host targets that modulate virus replication in mammalian cells in general. This protocol describes the development and validation of an assay for high-throughput RNAi screening in mammalian cells, the key considerations and preparation steps important for conducting a primary high-throughput RNAi screen, and a step-by-step guide for conducting a primary high-throughput RNAi screen; in addition, it broadly outlines the methods for conducting secondary screen validation and tertiary validation studies. The benefit of high-throughput RNAi screening is that it allows one to catalogue, in an extensive and unbiased fashion, host factors that modulate any aspect of virus replication for which one can develop an in vitro assay such as infectivity, burst size, and cytotoxicity. It has the power to uncover biotherapeutic targets unforeseen based on current knowledge.

Here we describe a protocol for employing high-throughput RNAi screening to uncover host targets that can be manipulated to enhance oncolytic virus therapy, specifically rhabodvirus and vaccinia virus therapy, but it can be readily adapted to other oncolytic virus platforms or for discovering host genes that modulate virus replication generally.

الفرز على نطاق الجينوم [رني] تحديد العوامل المضيفة التي تعدل علاج فيروس أونكوليتيك

High-throughput genome-wide RNAi (RNA interference) screening technology has been widely used for discovering host factors that impact&#160;virus ...

Cancer Research

A Method for Remotely Silencing Neural Activity in Rodents During Discrete Phases of Learning

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different forms of learning. Specifically, viral-mediated delivery is used to express a genetically modified inhibitory G-protein coupled receptor (the Designer Receptor) into a discrete brain region in the rodent. Three weeks later, when designer receptor expression levels are high, a pharmacological agent (the Designer Drug) is administered systemically 30 min prior to a specific behavioral session. The drug has affinity for the designer receptor and thus results in inhibition of neurons that express the designer receptor, but is otherwise biologically inert. The brain region remains silenced for 2-5 hr (depending on the dose and route of administration). Upon completion of the behavioral paradigm, brain tissue is assessed for correct placement and receptor expression. This approach is particularly useful for determining the contribution of individual brain regions to specific components of behavior and can be used across any number of behavioral paradigms.

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while rats are engaged in learning and memory tasks. The approach combines pharmacogenetics (Designer-Receptors-Exclusively-Activated-by-Designer-Drugs) with a behavioral paradigm (sensory preconditioning) that is designed to distinguish between different components of learning.

A طريقة لإسكات العصبية عن بعد آخر في القوارض وخلال مراحل متقطعة التعلم

This protocol describes how to temporarily and remotely silence neuronal activity in discrete brain regions while animals are engaged in learning and ...

Behavior

Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis. In recent years, oncolytic virotherapies have proven themselves to be potentially powerful tools to fight malignancy. Due to the unique dual blood supply in the liver, it is possible to apply therapies locally to orthotopic liver tumors, which are predominantly fed by arterial blood flow. We have previously demonstrated that hepatic arterial delivery of oncolytic viruses results in safe and efficient transduction efficiency of multifocal HCC lesions, resulting in significant prolongation of survival in immune competent rats. This procedure closely mimics the application of transarterial embolization in patients, which is the standard palliative care provided to many HCC patients. The ability to administer tumor therapies through the hepatic artery in rats allows for a highly sophisticated preclinical model for evaluating novel viral vectors under development. Here we describe the detailed protocol for microdissection of the hepatic artery for infusion of oncolytic virus vectors to treat orthotopic HCC.

Oncolytic virotherapies are under development as novel therapeutics for the treatment of hepatocellular carcinoma (HCC). Here we describe a method for locoregional therapy of HCC via hepatic arterial administration of oncolytic virus.

إدارة Transarterial من حال الورم فيروسات للعلاج العلاج الموضعي من مثلي HCC في الجرذان

Hepatocellular carcinoma (HCC) is a disease with limited treatment options and poor prognosis.  In recent years, oncolytic virotherapies have proven ...

Generation of Transgenic Rats using a Lentiviral Vector Approach

Transgenic animal models are fundamentally important for modern biomedical research. The incorporation of foreign genes into early mouse or rat embryos is an invaluable tool for gene function analysis in living organisms. The standard transgenesis method is based on microinjecting foreign DNA fragments into a pronucleus of a fertilized oocyte. This technique is widely used in mice but remains relatively inefficient and technically demanding in other animal species. The transgene can also be introduced into one-cell-stage embryos via lentiviral infection, providing an effective alternative to standard pronuclear injections, especially in species or strains with a more challenging embryo structure. In this approach, a suspension that contains lentiviral vectors is injected into the perivitelline space of a fertilized rat embryo, which is technically less demanding and has a higher success rate. Lentiviral vectors were shown to efficiently incorporate the transgene into the genome to determine the generation of stable transgenic lines. Despite some limitations (e.g., Biosafety Level 2 requirements, DNA fragment size limits), lentiviral transgenesis is a rapid and efficient transgenesis method. Additionally, using female rats that are mated with a fertile male strain with a different dominant fur color is presented as an alternative to generate pseudopregnant foster mothers.

This article aims to provide the methodology for lentiviral transgenesis in rat embryos using multiple injections of a virus suspension into the zygote perivitelline space. Female rats that are mated with a fertile male strain with a different dominant fur color is used to generate pseudopregnant foster mothers.

جيل من الفئران المعدلة وراثيا باستخدام نهج ناقلات لينديفيس

Transgenic animal models are fundamentally important for modern biomedical research. The incorporation of foreign genes into early mouse or rat embryos is ...

Bioengineering

Methods and Tips for Intravenous Administration of Adeno-associated Virus to Rats and Evaluation of Central Nervous System Transduction

Adeno-associated virus (AAV) vectors are a key reagent in the neurosciences for clustered regularly interspaced short palindromic repeats (CRISPR), optogenetics, cre-lox targeting, etc. The purpose of this manuscript is to aid the investigator attempting expansive central nervous system (CNS) gene transfer in the rat via tail vein injection of AAV. Wide-scale expression is relevant for conditions with widespread pathology, and a rat model is significant due to its greater size and physiologic similarities to humans compared to mice. In this example application, a wide-scale neuronal transduction is used to mimic a neurodegenerative disease that affects the entire spinal cord, amyotrophic lateral sclerosis (ALS). The efficient wide-scale CNS transduction can also be used to deliver therapeutic protein factors in pre-clinical studies. After a post-injection expression interval of several weeks, the effects of the transduction are evaluated. For a green fluorescent protein (GFP) control vector, the amount of GFP in the cerebellum is estimated quickly and reliably by a basic imaging program. For motor disease phenotypes that are induced by the ALS related protein transactive response DNA-binding protein of 43 kDa (TDP-43), the deficits are scored by escape reflex and rotarod. Beyond disease modeling and gene therapy, there are diverse potential applications for the wide-scale gene targeting described here. The expanded use of this method will aid in expediting hypothesis testing in the neurosciences and neurogenetics.

Methods for a wide-scale central nervous system gene delivery in the rat are covered. In this example, the purpose is to mimic a disease that affects the entire spinal cord. The widespread transduction can be used to deliver a therapeutic protein to the CNS from a one-time, peripheral administration.

طرق ونصائح للإدارة عن طريق الحقن الوريدي من الفيروسات المرتبطة بالغدد للفئران والتقييم لتوصيل الجهاز العصبي المركزي

Adeno-associated virus (AAV) vectors are a key reagent in the neurosciences for clustered regularly interspaced short palindromic repeats (CRISPR), ...

Neuroscience

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

نموذج الفئران من عدوى الدماغ EcoHIV

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...

Monitoring Endoplasmic Reticulum Calcium Homeostasis Using a Gaussia Luciferase SERCaMP

The endoplasmic reticulum (ER) contains the highest level of intracellular calcium, with concentrations approximately 5,000-fold greater than cytoplasmic levels. Tight control over ER calcium is imperative for protein folding, modification and trafficking. Perturbations to ER calcium can result in the activation of the unfolded protein response, a three-prong ER stress response mechanism, and contribute to pathogenesis in a variety of diseases. The ability to monitor ER calcium alterations during disease onset and progression is important in principle, yet challenging in practice. Currently available methods for monitoring ER calcium, such as calcium-dependent fluorescent dyes and proteins, have provided insight into ER calcium dynamics in cells, however these tools are not well suited for in vivo studies. Our lab has demonstrated that a modification to the carboxy-terminus of Gaussia luciferase confers secretion of the reporter in response to ER calcium depletion. The methods for using a luciferase based, secreted ER calcium monitoring protein (SERCaMP) for in vitro and in vivo applications are described herein. This video highlights hepatic injections, pharmacological manipulation of GLuc-SERCaMP, blood collection and processing, and assay parameters for longitudinal monitoring of ER calcium.

Monitoring Endoplasmic Reticulum Calcium Homeostasis Using a Gaussia Luciferase SERCaMP

Endoplasmic reticulum calcium homeostasis is disrupted in diverse pathologies. A secreted ER calcium monitoring protein (SERCaMP) reporter can be used to detect disruptions in the ER calcium store. This protocol describes the use of a Gaussia luciferase SERCaMP to examine ER calcium homeostasis in vitro and in vivo.

رصد الباطنة شبكية الكالسيوم باستخدام<em&gt; Gaussia</em&gt; luciferase المراسل SERCaMP

The endoplasmic reticulum (ER) contains the highest level of intracellular calcium, with concentrations approximately 5,000-fold greater than cytoplasmic ...

Biology

Viral Transgene Expression in Rodent Hearts and the Assessment of Cardiac Arrhythmia Risk

Heart disease is the leading cause of morbidity and mortality worldwide. Due to the ease of handling&#160;and abundance of transgenic strains, rodents have become essential models for cardiovascular research. However, spontaneous lethal cardiac arrhythmias that often cause mortality in heart disease patients are rare in rodent models of heart disease. This is primarily due to the species differences in cardiac electrical properties between human&#160;and rodents and poses a challenge to the study of cardiac arrhythmias using rodents. This protocol describes an approach to enable efficient transgene expression in mouse and rat ventricular myocardium using echocardiography-guided intramuscular injections of recombinant virus (adenovirus and adeno-associated virus). This work also outlines a method to enable reliable assessment of cardiac susceptibility to arrhythmias using isolated, Langendorff-perfused mouse and rat hearts with both adrenergic and programmed electrical stimulations. These techniques are critical for studying heart rhythm disorders associated with adverse cardiac remodeling after injuries, such as myocardial infarction.

The present protocol describes methods for transgene expression in rat and mouse hearts by direct intramyocardial injection of the virus under echocardiography guidance. Methods for the assessment of the susceptibility of hearts to ventricular arrhythmias by the programmed electrical stimulation of isolated, Langendorff-perfused hearts are also explained here.

التعبير الجيني المحورة الفيروسية في قلوب القوارض وتقييم مخاطر عدم انتظام ضربات القلب

Heart disease is the leading cause of morbidity and mortality worldwide. Due to the ease of handling&#160;and abundance of transgenic strains, rodents ...

Establishing an Active HIV Infection in a Rat Model

Source: Li, H., McLaurin, et al. <a href="https://app.jove.com/v/62137">A Rat Model of EcoHIV Brain Infection.</a> J. Vis. Exp. (2021).
This video demonstrates the procedure of injecting chimeric HIV viruses into the cortex of a rat&#39;s brain, where the virus RNA integrates into the host genome, replicates, and spreads to establish an active HIV infection in the experimental model.

تحديد عدوى فيروس نقص المناعة البشرية النشطة في نموذج الفئران

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

JoVE Encyclopedia of Experiments

Neuropathology

Infectious Diseases

Targeting Liver Tumors with Oncolytic Viruses via the Hepatic Artery

Source: Altomonte, J. et al., <a href="https://app.jove.com/v/53757">Transarterial Administration of Oncolytic Viruses for Locoregional Therapy of Orthotopic HCC in Rats.</a>&#160;J. Vis. Exp. (2016)
The video demonstrates locoregional delivery of oncolytic viruses through hepatic arterial injection as a therapeutic approach for hepatocellular carcinoma. These viruses selectively replicate in tumor cells, inducing cell lysis and releasing tumor cell-associated antigens. This process triggers immunogenic cell death, inducing an anti-tumor response.

التعامل مع الفأر القطن في الدراسات لتقييم ما قبل السريرية من حال الورم الفيروسات

Summary

Explore More Videos

Chapters in this video