Name: near infrared photoimmunotherapy for mouse models of pleural dissemination
Uploaded: 2021-02-09T12:30:00
Description: Watch this Scientific Journal Video about Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination at JoVE.com

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Alle in vivo-eksperimenter blev udført i overensstemmelse med vejledningen til pleje og brug af laboratoriedyrressourcer i Nagoya University Animal Care and Use Committee (godkendelse # 2017-29438, # 2018-30096, # 2019-31234, # 2020-20104). Seks uger gamle homozygote athymic nøgen mus blev købt og vedligeholdt på Animal Center of Nagoya University. Ved udførelse af proceduren hos mus blev de bedøvet med isofluran (indledning: 4-5%, vedligeholdelse 2-3%); poten blev presset med pincet for at bekræfte dybde...

<ol>
	<li>Xu, X., Lu, H., Lee, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+Infrared+Light+Triggered+Photo/Immuno-Therapy+Toward+Cancers.">Near Infrared Light Triggered Photo/Immuno-Therapy Toward Cancers.</a> Frontiers in Bioengineering and Biotechnology. 8, (2020).</li><li>Mitsunaga, M., 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=Cancer+cell-selective+in+vivo+near+infrared+photoimmunotherapy+targeting+specific+membrane+molecules.">Cancer cell-selective in vivo near infrared photoimmunotherapy targeting specific membrane molecules.</a> Nature Medicine. 17, 1685-1691 (2011).</li><li>Kobayashi, H., Choyke, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-Infrared+Photoimmunotherapy+of+Cancer.">Near-Infrared Photoimmunotherapy of Cancer.</a> Accounts of Chemical Research. 52, 2332-2339 (2019).</li><li>Oseroff, A. R., Ohuoha, D., Hasan, T., Bommer, J. C., Yarmush, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibody-targeted+photolysis:+Selective+photodestruction+of+human+T-cell+leukemia+cells+using+monoclonal+antibody-chlorin+e6+conjugates.">Antibody-targeted photolysis: Selective photodestruction of human T-cell leukemia cells using monoclonal antibody-chlorin e6 conjugates.</a> Proceedings of the National Academy of Sciences of the United States of America. 83, 8744-8748 (1986).</li><li>Mew, D., Wat, C. K., Towers, G. H., Levy, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy:+treatment+of+animal+tumors+with+tumor-specific+monoclonal+antibody-hematoporphyrin+conjugates.">Photoimmunotherapy: treatment of animal tumors with tumor-specific monoclonal antibody-hematoporphyrin conjugates.</a> Journal of Immunology. 130, 1473-1477 (1983).</li><li>Vrouenraets, M. B., 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=Development+of+meta-tetrahydroxyphenylchlorin-monoclonal+antibody+conjugates+for+photoimmunotherapy.">Development of meta-tetrahydroxyphenylchlorin-monoclonal antibody conjugates for photoimmunotherapy.</a> Cancer Research. 59, 1505-1513 (1999).</li><li>Goff, B. A., 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=Photoimmunotherapy+and+biodistribution+with+an+OC125-chlorin+immunoconjugate+in+an+in+vivo+murine+ovarian+cancer+model.">Photoimmunotherapy and biodistribution with an OC125-chlorin immunoconjugate in an in vivo murine ovarian cancer model.</a> British Journal of Cancer. 70, 474-480 (1994).</li><li>Sato, 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=Photoinduced+Ligand+Release+from+a+Silicon+Phthalocyanine+Dye+Conjugated+with+Monoclonal+Antibodies:+A+Mechanism+of+Cancer+Cell+Cytotoxicity+after+Near-Infrared+Photoimmunotherapy.">Photoinduced Ligand Release from a Silicon Phthalocyanine Dye Conjugated with Monoclonal Antibodies: A Mechanism of Cancer Cell Cytotoxicity after Near-Infrared Photoimmunotherapy.</a> ACS Central Science. 4, 1559-1569 (2018).</li><li>Sato, K., Nagaya, T., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+in+the+treatment+of+pleural+disseminated+NSCLC:+Preclinical+experience.">Near infrared photoimmunotherapy in the treatment of pleural disseminated NSCLC: Preclinical experience.</a> Theranostics. 5, 698-709 (2015).</li><li>Nishinaga, Y., 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=Targeted+Phototherapy+for+Malignant+Pleural+Mesothelioma:+Near-Infrared+Photoimmunotherapy+Targeting+Podoplanin.">Targeted Phototherapy for Malignant Pleural Mesothelioma: Near-Infrared Photoimmunotherapy Targeting Podoplanin.</a> Cells. 9, 1019 (2020).</li><li>Sato, 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=Near+infrared+photoimmunotherapy+prevents+lung+cancer+metastases+in+a+murine+model.">Near infrared photoimmunotherapy prevents lung cancer metastases in a murine model.</a> Oncotarget. 6, 19747-19758 (2015).</li><li>Sato, K., Nagaya, T., Mitsunaga, M., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near+infrared+photoimmunotherapy+for+lung+metastases.">Near infrared photoimmunotherapy for lung metastases.</a> Cancer Letters. 365, 112-121 (2015).</li><li>Isobe, Y., 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=Near+infrared+photoimmunotherapy+targeting+DLL3+for+small+cell+lung+cancer.">Near infrared photoimmunotherapy targeting DLL3 for small cell lung cancer.</a> EBioMedicine. 52, 102632 (2020).</li><li>Nakamura, Y., 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=Near+infrared+photoimmunotherapy+in+a+transgenic+mouse+model+of+spontaneous+epidermal+growth+factor+receptor+(EGFR)-expressing+lung+cancer.">Near infrared photoimmunotherapy in a transgenic mouse model of spontaneous epidermal growth factor receptor (EGFR)-expressing lung cancer.</a> Molecular Cancer Therapeutics. 16, 408-414 (2017).</li><li>Nagaya, 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=Near+infrared+photoimmunotherapy+with+avelumab,+an+anti-programmed+death-ligand+1+(PD-L1)+antibody.">Near infrared photoimmunotherapy with avelumab, an anti-programmed death-ligand 1 (PD-L1) antibody.</a> Oncotarget. 8, 8807-8817 (2017).</li><li>Sato, 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=Spatially+selective+depletion+of+tumor-associated+regulatory+T+cells+with+near-infrared+photoimmunotherapy.">Spatially selective depletion of tumor-associated regulatory T cells with near-infrared photoimmunotherapy.</a> Science Translational Medicine. 8, (2016).</li><li>Sato, 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=Comparative+effectiveness+of+light+emitting+diodes+(LEDs)+and+lasers+in+near+infrared+photoimmunotherapy.">Comparative effectiveness of light emitting diodes (LEDs) and lasers in near infrared photoimmunotherapy.</a> Oncotarget. 7, 14324-14335 (2016).</li><li>Sato, K., Choyke, P. L., Kobayashi, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoimmunotherapy+of+Gastric+Cancer+Peritoneal+Carcinomatosis+in+a+Mouse+Model.">Photoimmunotherapy of Gastric Cancer Peritoneal Carcinomatosis in a Mouse Model.</a> PLoS One. 9, 113276 (2014).</li><li>McLemore, T. L., 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=Comparison+of+intrapulmonary,+percutaneous+intrathoracic,+and+subcutaneous+models+for+the+propagation+of+human+pulmonary+and+nonpulmonary+cancer+cell+lines+in+athymic+nude+mice.">Comparison of intrapulmonary, percutaneous intrathoracic, and subcutaneous models for the propagation of human pulmonary and nonpulmonary cancer cell lines in athymic nude mice.</a> Cancer Research. 48, 2880-2886 (1988).</li><li>Manzotti, C., Audisio, R. A., Pratesi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Importance+of+orthotopic+implantation+for+human+tumors+as+model+systems:+relevance+to+metastasis+and+invasion.">Importance of orthotopic implantation for human tumors as model systems: relevance to metastasis and invasion.</a> Clinical &amp; Experimental Metastasis. 11, 5-14 (1993).</li><li>Lwin, T. M., Hoffman, R. M., Bouvet, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advantages+of+patient-derived+orthotopic+mouse+models+and+genetic+reporters+for+developing+fluorescence-guided+surgery.">Advantages of patient-derived orthotopic mouse models and genetic reporters for developing fluorescence-guided surgery.</a> Journal of Surgical Oncology. 118, 253-264 (2018).</li><li>Sordat, B. C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> From Ectopic to Orthotopic Tumor Grafting Sites: Evidence for a Critical Role of the Host Tissue Microenvironment for the Actual Expression of the Malignant Phenotype. , 43-53 (2017).</li><li>Sato, 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=Photoimmunotherapy:+comparative+effectiveness+of+two+monoclonal+antibodies+targeting+the+epidermal+growth+factor+receptor.">Photoimmunotherapy: comparative effectiveness of two monoclonal antibodies targeting the epidermal growth factor receptor.</a> Molecular Oncology. 8, 620-632 (2014).</li><li>Nakajima, 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=The+effects+of+conjugate+and+light+dose+on+photo-immunotherapy+induced+cytotoxicity.">The effects of conjugate and light dose on photo-immunotherapy induced cytotoxicity.</a> BMC Cancer. 14, 389 (2014).</li><li>Nagaya, 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=Near+infrared+photoimmunotherapy+of+B-cell+lymphoma.">Near infrared photoimmunotherapy of B-cell lymphoma.</a> Molecular Oncology. 10, 1404-1414 (2016).</li><li>Sato, 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=Near+infrared+photoimmunotherapy+in+the+treatment+of+disseminated+peritoneal+ovarian+cancer.">Near infrared photoimmunotherapy in the treatment of disseminated peritoneal ovarian cancer.</a> Molecular Cancer Therapeutics. 14, 141-150 (2015).</li><li>Colin, D. J., Bejuy, O., Germain, S., Triponez, F., Serre-Beinier, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implantation+and+monitoring+by+pet/ct+of+an+orthotopic+model+of+human+pleural+mesothelioma+in+athymic+mice.">Implantation and monitoring by pet/ct of an orthotopic model of human pleural mesothelioma in athymic mice.</a> Journal of Visualized Experiments. 2019, (2019).</li><li>Opitz, I., 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=Local+recurrence+model+of+malignant+pleural+mesothelioma+for+investigation+of+intrapleural+treatment.">Local recurrence model of malignant pleural mesothelioma for investigation of intrapleural treatment.</a> European Journal of Cardio-Thoracic Surgery. 31, 772-778 (2007).</li><li>Bunn, P. A., Kelly, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+chemotherapeutic+agents+prolong+survival+and+improve+quality+of+life+in+non-small+cell+lung+cancer:+a+review+of+the+literature+and+future+directions.">New chemotherapeutic agents prolong survival and improve quality of life in non-small cell lung cancer: a review of the literature and future directions.</a> Clinical Cancer Research. 4, 1087-1100 (1998).</li><li>Astoul, P., Wang, X., Hoffman, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patient-like+nude-mouse+and+scid-mouse+models+of+human+lung+and+pleural+cancer+(review).">Patient-like nude-mouse and scid-mouse models of human lung and pleural cancer (review).</a> International Journal of Oncology. 3, 713-718 (1993).</li><li>Yamaguchi, H., Pantarat, N., Suzuki, T., Evdokiou, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-infrared+photoimmunotherapy+using+a+small+protein+mimetic+for+HER2-overexpressing+breast+cancer.">Near-infrared photoimmunotherapy using a small protein mimetic for HER2-overexpressing breast cancer.</a> International Journal of Molecular Sciences. 20, (2019).</li><li>Jing, H., 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=Imaging+and+selective+elimination+of+glioblastoma+stem+cells+with+theranostic+Near-Infrared-Labeled+CD133-Specific+antibodies.">Imaging and selective elimination of glioblastoma stem cells with theranostic Near-Infrared-Labeled CD133-Specific antibodies.</a> Theranostics. 6, 862-874 (2016).</li><li>Burley, T. A., 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=Near-infrared+photoimmunotherapy+targeting+EGFR-Shedding+new+light+on+glioblastoma+treatment.">Near-infrared photoimmunotherapy targeting EGFR-Shedding new light on glioblastoma treatment.</a> International Journal of Cancer. 142, 2363-2374 (2018).</li><li>Nagaya, 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=Near+infrared+photoimmunotherapy+using+a+fiber+optic+diffuser+for+treating+peritoneal+gastric+cancer+dissemination.">Near infrared photoimmunotherapy using a fiber optic diffuser for treating peritoneal gastric cancer dissemination.</a> Gastric Cancer. 22, 463-472 (2019).</li><li>Nagaya, 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=Endoscopic+near+infrared+photoimmunotherapy+using+a+fiber+optic+diffuser+for+peritoneal+dissemination+of+gastric+cancer.">Endoscopic near infrared photoimmunotherapy using a fiber optic diffuser for peritoneal dissemination of gastric cancer.</a> Cancer Science. 109, 1902-1908 (2018).</li><li>Harada, 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=Near-infrared+photoimmunotherapy+with+galactosyl+serum+albumin+in+a+model+of+diffuse+peritoneal+disseminated+ovarian+cancer.">Near-infrared photoimmunotherapy with galactosyl serum albumin in a model of diffuse peritoneal disseminated ovarian cancer.</a> Oncotarget. 7, 79408-79416 (2016).</li><li>Journals, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=JNCI+Journal+of+the+National+Cancer+Institute+Way+to+Better+DNA.">JNCI Journal of the National Cancer Institute Way to Better DNA.</a> Annals of Internal Medicine. 37, 1-9 (2008).</li></ol>

I denne undersøgelse demonstrerede vi en metode til måling af NIR-PIT's terapeutiske virkning på den pleurale formidlingsmodel af MPM. Meget selektiv celledrab blev udført med NIR-PIT; således blev det normale væv næppe beskadiget23,24,25. Med denne type selektiv celleaflivning blev NIR-PIT påvist at være sikker i formidlede modeller9,...

Kræft er fortsat en af de førende årsager til dødelighed på trods af årtiers forskning. En af grundene er, at strålebehandling og kemoterapi er meget invasive teknikker, som kan begrænse deres terapeutiske fordele. Cellulære- eller molekylære-målrettede behandlinger, som er mindre invasive teknikker, får øget opmærksomhed. Fotoimmunoterapi er en behandlingsmetode, der synergistisk forbedrer den terapeutiske virkning ved at kombinere immunterapi og lysbehandling. Immunterapi øger tumorimmunitet ved at øge immunogeniciteten af tumormikromiljøet og reducere immunreguleringsundertrykkelse, hvilket resulterer i ødelæggelse af tumorer i kroppen. Lysbehandling ødelægger primære tumorer med en kombination af photosensitizers og lysstråler, og tumor-specifikke antigener frigivet fra tumorceller øge tumor immunitet. Tumorer kan selektivt behandles ved hjælp af photosensitizers, da de er specifikke og selektive for målcellerne. Lysterapiens modalitet omfatter fotodynamisk terapi (PDT), fototermisk terapi (PTT) og fotokemibaserede behandlinger1.
Nær-infrarød fotoimmunoterapi (NIR-PIT) er en nyligt udviklet metode til antitumor lysbehandling, der kombinerer fotokemisk-baseret terapi og immunterapi1,2. NIR-PIT er en molekylært målrettet behandling, der er rettet mod specifikke celleoverflademolekyler gennem bøjning af et nær-infrarødt siliciumphthalocyaninfarvestof, IRdye 700DX (IR700), til et monoklonalt antistof (mAb). Målcellens cellemembran ødelægges ved bestråling med NIR-lys (690 nm)3.
Konceptet med at bruge målrettet lysterapi ved at kombinere konventionelle fotoensibilisatorer og antistoffer eller målrettet PDT er over tre årtier gammel4,5. Tidligere undersøgelser har forsøgt at målrette konventionelle PDT-midler ved at konjugatere dem til antistoffer. Der var dog begrænset succes, fordi disse konjugates blev fanget i leveren på grund af hydrofobicity af photosensitizers6,7. Desuden er NIR-PIT's mekanisme helt forskellig fra den konventionelle PDT's. Konventionelle photosensitizers generere oxidativ stress, der skyldes en energi konvertering, der absorberer lys energi, forvredes til en ophidset tilstand, overgange til jorden tilstand, og forårsager apoptose. Men NIR-PIT forårsager hurtig nekrose ved direkte at ødelægge cellemembranen ved at aggregere photosensitizers på membranen gennem en fotokemisk reaktion8. NIR-PIT er bedre end konventionelle målrettede PDT på mange måder. Konventionelle fotoensibilisatorer har lave udryddelseskoefficienter, der kræver fastgørelse af et stort antal photosensitizers til et enkelt antistofmolekyle, hvilket potentielt reducerer bindende affinitet. De fleste konventionelle fotoensibilisatorer er hydrofobiske, hvilket gør det vanskeligt at binde photosensitizers til antistoffer uden at kompromittere deres immunreaktivitet eller in vivo målakkumulering. Konventionelle fotoensibilisatorer absorberer typisk lys i det synlige område, hvilket reducerer vævspenetration.
Flere undersøgelser af NIR-PIT rettet mod intrathoraciske tumorer som lungekræft og maligne pleuralmesotheliomceller (MPM) er blevet rapporteret9,10,11,12,13,14,15,16,17. Det er dog kun nogle få rapporter , der har beskrevet effekten af NIR-PIT i pleuralformidlet MPM - eller lungekræftmodellerne9,10,11,12. Subkutane tumor xenograft modeller menes at være standard tumor modeller og er i øjeblikket meget udbredt til at evaluere antitumor virkninger af nye behandlinger18. Det subkutane tumormikromiljø er dog ikke eftergivende for udviklingen af en passende vævsstruktur eller en tilstand, der korrekt opsummerer en ægte ondartet fænotype19,20,21,22. Ideelt set bør ortotopiske sygdomsmodeller etableres for en mere præcis vurdering af antitumoreffekterne.
Her demonstrerer vi en metode til effektevaluering i en musemodel af pleuralformidlet lungekræft, som blev behandlet ved hjælp af NIR-PIT. En pleural formidling mus model genereres ved at injicere tumorceller i brysthulen og bekræftet ved hjælp af luciferase luminescens. Musen blev behandlet med en intravenøs injektion af mAb konjugeret med IR700 og NIR bestråling til brystet. Den terapeutiske virkning blev evalueret ved hjælp af luciferase luminescens.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.25w/v% Trypsin-1mmol/l EDTA 4Na Solution with Phenol Red</td>
			<td>Wako</td>
			<td>209-016941</td>
			<td>for cell culture</td>
		</tr>
		<tr>
			<td>1mL syringe</td>
			<td>TERUMO</td>
			<td>SS-01T</td>
			<td>for mice experiment</td>
		</tr>
		<tr>
			<td>30G needle</td>
			<td>Nipro</td>
			<td>1907613</td>
			<td>for mice experiment</td>
		</tr>
		<tr>
			<td>BALB/cSlc-nu/nu</td>
			<td>Japan SLC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Collidal Blue Staining Kit</td>
			<td>Invitrogen</td>
			<td>LC6025</td>
			<td>use for gel protein staining</td>
		</tr>
		<tr>
			<td>Coomassie (bradford) Plus protein assay</td>
			<td>Thermo Fisher Scientific Inc (Waltham, MA, USA)</td>
			<td>PI-23200</td>
			<td>for measuring the APC concentration</td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide (DMSO)</td>
			<td>Wako</td>
			<td>043-07216</td>
			<td>use for conjugation of IR700</td>
		</tr>
		<tr>
			<td>D-Luciferin (potassium salt)</td>
			<td>Cayman Chemical</td>
			<td>14681</td>
			<td>for bioluminescence imaging and DLIT</td>
		</tr>
		<tr>
			<td>GraphPad Prism7</td>
			<td>GraphPad software</td>
			<td></td>
			<td>for statistical analysis</td>
		</tr>
		<tr>
			<td>Image Studio</td>
			<td>Li-Cor Biosciences</td>
			<td></td>
			<td>for analyzing 700 nm fluorescent image</td>
		</tr>
		<tr>
			<td>IRDye 700DX Ester Infrared Dye</td>
			<td>LI-COR Bioscience (Lincoln, NE, USA)</td>
			<td>929-70011</td>
			<td></td>
		</tr>
		<tr>
			<td>isoflurane</td>
			<td>Wako</td>
			<td>095-06573</td>
			<td>for mice anesthesia</td>
		</tr>
		<tr>
			<td>IVIS Spectrum CT</td>
			<td>PerkinElmer</td>
			<td></td>
			<td>for capturing bioluminescent image and DLIT</td>
		</tr>
		<tr>
			<td>Living Image</td>
			<td>PerkinElmer</td>
			<td></td>
			<td>for analyzing bioluminescent image and DLIT</td>
		</tr>
		<tr>
			<td>Na2HPO4</td>
			<td>SIGMA-ALDRICH (St. Louis, MO, USA)</td>
			<td>S9763</td>
			<td>use for conjugation of IR700</td>
		</tr>
		<tr>
			<td>NIR Laser</td>
			<td>Changchun New Industries Optoelectronics Technology</td>
			<td>MRL-III-690R</td>
			<td>for NIR irradiation</td>
		</tr>
		<tr>
			<td>Novex WedgeWell 4 to 20%, Tris-Glycine, 1.0 mm, Mini Protein Gel, 12 well</td>
			<td>Invitrogen</td>
			<td>XP04202BOX</td>
			<td>use for SDS-PAGE</td>
		</tr>
		<tr>
			<td>NuPAGE LDS Sample Buffer (x4)</td>
			<td>Invitrogen</td>
			<td>NP0007</td>
			<td>use for SDS-PAGE</td>
		</tr>
		<tr>
			<td>Optical power meter</td>
			<td>Thorlabs (Newton, NJ, USA)</td>
			<td>PM100</td>
			<td>for measuring the output of the NIR laser&#160;</td>
		</tr>
		<tr>
			<td>PBS(-)</td>
			<td>Wako</td>
			<td>166-23555</td>
			<td></td>
		</tr>
		<tr>
			<td>Pearl Trilogy imaging system</td>
			<td>Li-Cor Biosciences</td>
			<td></td>
			<td>for capturing 700 nm fluorecent image</td>
		</tr>
		<tr>
			<td>Penicilin-Streptomycin Solution (x100)</td>
			<td>Wako</td>
			<td>168-23191</td>
			<td>for cell culture</td>
		</tr>
		<tr>
			<td>Puromycin Dihydrochloride</td>
			<td>ThermoFisher</td>
			<td>A1113803</td>
			<td>for luciferase transfection</td>
		</tr>
		<tr>
			<td>RediFect Red-Fluc-Puromycin Lentiviral Prticles</td>
			<td>PerkinElmer</td>
			<td>CLS960002</td>
			<td>for luciferase transfection</td>
		</tr>
		<tr>
			<td>RPMI-1640 with L-glutamine and Phenol Red</td>
			<td>Wako</td>
			<td>189-02025</td>
			<td>for cell culture</td>
		</tr>
		<tr>
			<td>Sephadex G25 column (PD-10)&#160;</td>
			<td>GE Healthcare (Piscataway, NJ, USA)</td>
			<td>17-0851-01</td>
			<td>use for conjugation of IR700</td>
		</tr>
		<tr>
			<td>UV-1900i</td>
			<td>Shimadzu</td>
			<td></td>
			<td>for measuring the APC concentration</td>
		</tr>
	</tbody>
</table>

near infrared photoimmunotherapy for mouse models of pleural dissemination

Effekten af fotoimmunoterapi kan evalueres mere præcist med en ortotopisk musemodel end med en subkutan. En pleural formidlingsmodel kan anvendes til evaluering af behandlingsmetoder for intrathoraciske sygdomme som lungekræft eller ondartet lungehindekræft.
Nær-infrarød fotoimmunotherapy (NIR-PIT) er en nyligt udviklet kræftbehandling strategi, der kombinerer specificiteten af tumor-målretning antistoffer med toksicitet forårsaget af en photoabsorber (IR700Dye) efter eksponering for NIR lys. Effekten af NIR-PIT er blevet rapporteret ved hjælp af forskellige antistoffer; Men kun få rapporter har vist den terapeutiske virkning af denne strategi i en ortotopisk model. I denne undersøgelse viser vi et eksempel på effektevaluering af den pleuralformidlet lungekræftmodel, som blev behandlet ved hjælp af NIR-PIT.

Anti-podoplanin antistof NZ-1 blev konjugeret med IR700 til at generere NZ-1-IR700. Vi bekræftede bindingen af NZ-1 og IR700 på en SDS-PAGE (figur 8). Luciferase-udtrykke H2373 (H2373-luc) blev udarbejdet ved transfecting maligne lungehindekræft celler (H2373) med en luciferase gen10.
Vi bedøvede 8-12 uger gamle kvindelige homozygote athymiske nøgenmus og injicerede 1 × 105 H2373-luc celler i brysthulen. Dagen for injektion...

Watch this Scientific Journal Video about Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination at JoVE.com

Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination

Nær-infrarød fotoimmunotherapy (NIR-PIT) er en spirende kræft terapeutisk strategi, der udnytter et antistof-photoabsorber (IR700Dye) konjugat og NIR lys til at ødelægge kræftceller. Her præsenterer vi en metode til at evaluere antitumoreffekten af NIR-PIT i en musemodel af pleuralformidlet lungekræft og ondartet lungehindekræft ved hjælp af bioluminescensbilleddannelse.

I nærheden af infrarød fotoimmunoterapi til musemodeller af pleural formidling

The efficacy of photoimmunotherapy can be evaluated more accurately with an orthotopic mouse model than with a subcutaneous one. A pleural dissemination ...

Denne metode kan bruges til at evaluere de terapeutiske virkninger af NIR-PIT på thorax tumorer i en klinisk relevant tumormodel. NIR-PIT-procedurerne ved hjælp af den pleuralformidlet kræftmodel er lette at forstå og udføre. For at oprette en museformidlingsmodel skal du bruge polystyrenskum til at lave en prop og desinficere den med 70% ethanol.Fastgør nålen til proppen med spidsen strækker sig fem millimeter ud af proppen. Bøj spidsen af en 30-gauge nål med rene sammentak eller mod en hård genstand rengøres med 70% ethanol, og fyld en sprøjte med en gange 10 til de seks mål tumorceller i 100 mikroliters af PBS. Bekræft en manglende reaktion på pedalrefleks i en bedøvet, 19 til 21 gram, otte til 12 uger gammel, kvinde, homozygote, athymisk nøgen mus.Sæt nålen ind i brystet gennem intercostal rummet, flytte nålen op og ned for at undgå at kontakte ribbenene. Når spidsen er gået gennem interkostalrummet, skal sprøjten placeres, så den presses mod musen, og injicere hele mængden af målceller. Efter injektionen rulle musen to til tre gange for at sprede cellerne i hele brysthulen, og returnere musen til sit bur med overvågning indtil fuld helbredelse.24 timer efter celle injektion og hver dag derefter, injicere bedøvet tumor celle-injicerede mus med 200 mikroliter på 15 milligram per milliliter af D-luciferin. Efter 10 minutter skal musene placeres i en bioluminescensbilled og åbne Acquisition Control Panel i billedsoftwaren. Vælg Selvlysende, Fotografi og Overlejring.Indstil eksponeringstiden til Auto, Binning til Small, F/Stop til en for Luminescent og til otte for Fotografi, og feltet vis til C.Click Erhverve at billedet bioluminescens. Angiv visningsformatet til radianer, og vælg cirklen i værktøjerne område af interesse i panelet Værktøjspalette. Klik på Mål områder af interesse for at måle overfladens bioluminescerende intensitet, og brug Konfigurer måling til at vælge de værdier, der er relevante for eksperimentet.Eksporter denne datatabel som en csv-fil. Brug derefter de samlede fluxværdier for den bioluminescerende intensitet kvantificering i filen. Før du udfører nær-infrarød lysbehandling af tumor-injicerede mus, skal du bruge en effektmåler til at måle lysdosis af en 690-nanometer bølgelængde laser, og justere produktionen til 100 milliwatt per kvadratcentimeter.24 timer før behandlingen injicerede intravenøst 100 mikrogram antistoffotosensibilisatorkonjugat i 50 til 200 mikroliter pbs via halevenen på det tumorinst injicerede dyr. På dagen for lysbehandling behandling, placere bedøvet konjugat-injiceret, tumor-laden mus i liggende position, og skjold de ikke-mål steder med aluminiumsfolie. Når alle skjoldene er placeret, skal du bruge en 100-joules-per-kvadrat-centimeter laser til at bestråle brysthulen med nær-infrarødt lys i ca. 30 sekunder.Når bestrålingen er færdig, og musen er vågnet, skal dyret vende tilbage til buret og måle bioluminescensen dagligt som demonstreret. I denne repræsentative analyse blev bøjningen af anti-podoplanin antistof med IR700 bekræftet ved SDS-PAGE-analyse. Efter tumorcelleindsprøjtning skal bioluminescensbilleddannelse og diffus luminescensbilleddannelsestomografi udføres for at bestemme, hvilke mus der udtrykker tilstrækkelig luciferaseaktivitet i brysthulen til yderligere undersøgelse.På dag fem efter injektion demonstrerer anti-podoplanin antistof IR700-injicerede mus høj IR700 fluorescens og luciferaseaktivitet i thoraxtumorer, hvilket indikerer, at intravenøst injiceret IR700-konjugeret antistof når formidlet pleural tumorsteder. Især demonstrerer pleuralformidlet mus behandlet med nær-infrarød fotoimmunoterapi en nedsat luciferaseaktivitet, mens de relative lysenheder i kontrolgruppen udviser en gradvis stigning i intensiteten. Da den krævede NIR-bestrålingsenergi afhænger af cellelinjen og antistofferne, skal du sørge for at kontrollere forholdene på forhånd in vitro.

Research

JoVE Journal

Medicine

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

Near Infrared Optical Projection Tomography for Assessments of &beta;-cell Mass Distribution in Diabetes Research

Near Infrared Optical Projection Tomography for Assessments of ÃƒÅ½Ã‚Â²-cell Mass Distribution in Diabetes Research

Near Infrared Optical Projection Tomography for Vurderinger af β-celle Mass Distribution i Diabetes Research

By adapting OPT to include the capability of imaging in the near infrared (NIR) spectrum, we here illustrate the possibility to image larger bodies of ...

Mouse Models for Graft Arteriosclerosis

Musemodeller for Graft Åreforkalkning

Graft arteriosclerois (GA),  also called allograft vasculopathy, is a pathologic lesion that develops over  months to years in transplanted organs ...

Tissue-simulating Phantoms for Assessing Potential Near-infrared Fluorescence Imaging Applications in Breast Cancer Surgery

Tissue-simulering Phantoms til vurdering af potentielle nærinfrarøde fluorescensimagografi Applications i Breast Cancer Surgery

Inaccuracies in intraoperative tumor localization and evaluation of surgical margin status result in suboptimal outcome of breast-conserving surgery ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Mus nyretransplantation: modeller af allotransplantatafstødning

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

Højfrekvente Ultrasound Imaging af Mouse Cervikal lymfeknuder

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal ...

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

Kombineret Nær-infrarød Fluorescent Imaging og Micro-computertomografi for Direkte Visualisering Cerebral tromboemboli

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of ...

Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice

Bioluminescens og Nær-infrarød Imaging af optisk neuritis og hjernebetændelse i EAE model af multipel sklerose i mus

Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing ...

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy

Skeletmuskulatur neurovaskulære kobling, Oxidative kapacitet og mikrovaskulære funktion med 'Et Stop Shop' nær-infrarød spektroskopi

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic ...

A New Best Practice for Validating Tail Vein Injections in Rat with Near-infrared-Labeled Agents

En ny bedste praksis for validering hale vene injektioner i rotte med nær-infrarød-mærket agenter

Intravenous (IV) administration of agents into the tail vein of rats can be both difficult and inconsistent. Optimizing tail vein injections is a key part ...