Wir danken für die Unterstützung aus dem National Cancer Institute (NCI) R01 CA128908 und Stanford Medical Scholar Research Fellowship. Kein anderer potenzieller Interessenkonflikt relevanten zu diesem Artikel berichtet wurde.

Ein. Tumormodell <ol><li> Kultur H460-Zellen (American Type Culture Collection) in RPMI 1640 Medium mit 10% fetalem Rinderserum und 1% Penicillin / Streptomycin (Invitrogen Life Technologies) ergänzt. Es sei darauf hingewiesen, dass die Auswahl von Zelllinien, Kulturmedien, Standorte der Inokulation Anzahl von Xenotransplantaten pro Maus und anderen Erwägungen alle zu den Zielen der Studie einer bestimmten zugeschnitten werden sollen. Hier werden wir nur präsentieren ein bestimmtes Projekt Design als Illustration dienen. </li><li> Pflegen Zelllinien in einer befeuchteten Atmosphäre von 5% CO 2 bei 37 ° C und Veränderung auf frisches Medium jeden zweiten Tag. </li><li> Wenn ein 75% konfluente Monoschicht von Zellen gebildet wird, lösen die Monoschicht mit Trypsin und distanzieren Zellen in eine Einzelzell-Suspension für weitere Zellkultur. </li><li> Auszusetzen ungefähr 1 × 10 6 H460 Zellen in Phosphat-gepufferter Kochsalzlösung (PBS; Invitrogen) und Implantat subkutan indie linken und rechten Schultern des nackten Mäusen (weiblich athymische Nacktmäuse (nu / nu), 4 - 6 Wochen alt, Charles River Laboratories, Inc.). </li><li> Lassen Tumoren auf 150 wachsen - 200 mm 3. Es dauert etwa 2 Wochen für H460 Tumorxenotransplantaten auf diese Größe zu wachsen. Standard Bremssattel Messung erfolgt nach Tumorgrößen verfolgen durchgeführt. </li><li> Wenn Tumore die ideale Größe der Tumor tragenden Mäusen sind nun bereit für die Behandlung und in-vivo-Bildgebung sowohl über PET und CLI zu erreichen. </li></ol> 2. PET <ol><li> Führen Sie die PET-Untersuchungen nach diesem Zeitplan oder eine Variation davon abhängig vom konkreten Projekt (Abbildung 1) 12. Eine Anzahl von Faktoren können Einfluß auf die Gestaltung des Zeitplans, einschließlich, aber nicht beschränkt auf, die Wahl der Tumorxenotransplantat Zelllinien, Antikrebsmittel und Dosierungsschemata begrenzt. Hier werden wir nur präsentieren eine spezifische Bildgebung Zeitplan. Die CLI-Studien sind nach der durchgeführt werdengleichen Zeitplan wie die der PET-Studien mit CLI durchgeführt unmittelbar nach dem entsprechenden PET. Es sollte auch beachtet werden, dass hier der Zweck der PET-Studien hauptsächlich für die Validierung der Ergebnisse CLI ist. Für normale Benutzer, die gerade wollen OI Instrumente für die Bildgebung radioaktiv markierten Sonden zu verwenden, ist keine PET notwendig. Allerdings, wenn ein Wunsch PET Validierung tut es sollte betont werden, dass PET und CLI Instrumente in unmittelbarer Nähe zur Validierung muss entfernt werden, damit aufgrund der kurzen Halbwertszeit von 18 F (109,77 min) erfolgreich zu sein. </li><li> Teilen Sie Mäuse in Behandlungs-und Kontrollgruppen (n ≥ 3 Stück). Saures Mäuse in der Behandlungsgruppe mit zwei Injektionen von Bevacizumab von 20 mg / kg an den Tagen 0 und 2. Tag 0 durch die erste Einspritzung definiert. Beachten Sie, dass an Tag -1 a Vorabtastung über beide PET und CLI durchgeführt werden. </li><li> Kleintier-PET von Tumor-tragenden Mäusen mit einem R4 Nagermodell Scanner (Siemens Medical Solutions USA durchgeführt werdenA, Inc.). </li><li> Anästhesieren alle Mäuse mit 2% Isofluran (Aerrane; Baxter) und injizieren mit 3&#39;-Desoxy-3&#39;-fluorthymidin-18 F (18 F-FLT; 7,3 bis 8,0 MBq [198 bis 215 uCi]) über die Schwanzvene. Das PET-Sonde ist, um vor der Injektion in PBS verdünnt werden. </li><li> Nach 1 Stunde, betäuben Mäusen wieder und stellen narkotisierten Mäusen neigen und in der Nähe der Mitte des Sichtfeldes der Kleintier-PET-Scanner. </li><li> Erhalten dreiminütigen statischen Scans und rekonstruieren die Bilder durch einen 2-dimensionalen geordneten Teilmengen-Erwartung maximale Algorithmus. Hintergrund Korrektur nicht erforderlich ist. </li><li> Zeichnen Regions of Interest (ROIs; 5 Pixel für koronalen und transaxial Scheiben) über den Tumoren auf den Zerfall korrigiert Ganzkörper-koronalen Bildern. Erhalten der maximalen Zählwerte pro Pixel pro Minute von der ROIs und Konvertieren in Zählungen pro Milliliter pro Minute durch die Verwendung einer Eichkonstante. Unter der Annahme eines Gewebes Dichte von 1 g / ml, wandeln die ROIs um Zählungenpro Gramm pro Minute. Bild bestimmen ROI-derived% ID / g Werten durch Dividieren Zählungen pro Gramm pro Minute durch injizierten Dosis. Schwächungskorrektur ist nicht notwendig. </li></ol> 3. CLI <ol><li> CLI ist mit einem IVIS Spectrum System (Caliper Life Sciences) durchgeführt werden. Erfassung und Analyse von Bildern sind durchgeführt werden, indem Wohnfläche Image 3.0 Software (Caliper Life Sciences). Wellenlängen aufgelösten spektralen Bildgebung ist es, unter Verwendung eines 18-set schmalbandige Emission Filter (490 - 850 nm) werden. Wiederum für jede Maus, führen CLI unmittelbar nach PET, um die Menge des radioaktiven Zerfalls zu minimieren, wenn die PET-Studien in dem Protokoll enthalten sind. </li><li> Platzieren Tiere in einer lichtdichten Kammer unter Isofluran-Narkose. Mehrere Mäuse können gleichzeitig platziert werden, um den Durchsatz zu erhöhen. </li><li> Erwerben Sie Bilder mit 3 min Belichtungszeit (f / Stop = 1, Binning = 4). Verwenden Sie die gleiche Beleuchtung Einstellungen (Lampenspannung, Filter, f / stop, Sichtfelder, binning) zum Erwerb sämtlicher Bilder. Verwenden Sie die dorsale Hautpartie um die Signalintensität Hintergrund Gewebe berechnen. Normalisieren Fluoreszenzemission Photonen pro Sekunde pro Zentimeter pro Steradiant (p / s / cm 2 / sr) quadriert. </li></ol> 4. Repräsentative Ergebnisse Visuellen Vergleich zwischen CLI und PET-Bilder können leicht durchgeführt werden. Nach Vereinheitlichung der Maßstab auf Bilder von derselben Modalität und Ort CLI-und PET-Bilder nebeneinander kann man in diesem repräsentativen Panel (2A), dass sowohl CLI und PET deutlich zurückgegangen Signale von H460 Xenotransplantate in behandelten Mäusen von der Vorbehandlung ergab sehen bis Tag 3, was darauf hindeutet signifikanten therapeutischen Effekt. Als Vergleich wurden mäßig bis unverändert Signalen erhöht in unbehandelten Mäusen während der gleichen Zeitperiode (Daten nicht gezeigt) beobachtet. Durch visuelle Inspektion allein kann man beobachten, dass es eine gute Konsistenz zwischen Tumor Kontraste, die visuell sindIzed von CLI und PET. In der Tat hat diese visuelle Korrelation ausreichende Auflösung der zentralen Nekrose des Tumors zweitrangig gegenüber der Anti-Krebs-Therapie (Bitte vergleichen Sie die CLI-und PET-Bilder von Tag 3) zeigen. Um die bildgebende Befunde Quantifizierungen validieren und Korrelationsanalyse durchgeführt werden kann. Quantifizierungen von CLI und PET-Bilder und eine einfache Montage durch lineare Regression ergab, dass die zwei Modalitäten Tat hatte eine ausgezeichnete Korrelation (2B, R 2 = 0,9309 für die 18 F-FLT sondiert Behandlungsgruppe). Bemerkenswert ist, in all unserer CLI-und PET-Bildgebung mit verschiedenen Tumor-Modellen und anderen Krebsmedikamenten die Steigungen der Anfälle sind auch bemerkenswert nahe, was darauf hindeutet, eine hervorragende Passform der linearen Regression sogar aller Daten conglomerated (Daten nicht gezeigt). Beide repräsentative Bilder von unserem früheren Veröffentlichung 12 angepasst. er.within-page = &quot;always&quot;&gt; <img src="/files/ftp_upload/4341/4341fig1.jpg" alt="Abbildung 1"/> Abbildung 1. Schematische Darstellung des experimentellen Design von PET und CLI Studien. Tumoren wurden bilateral im Schulterbereich implantiert und läßt auf 150-200 mm 3 wachsen und tumortragenden Mäuse wurden in vivo Bildgebung mittels PET und CLI unterworfen am Tag -1, 1, und 3. Bevacizumab Behandlung wurde durch zwei Injektionen von 20 mg / kg an den Tagen 0 und 2 durchgeführt wird. <img src="/files/ftp_upload/4341/4341fig2.jpg" alt="Abbildung 2"/> Abbildung 2. (A) In-vivo-CLI und PET-Bilder von Mäusen, die mit H460 Xenotransplantate Bevacizumab vor der Behandlung (Pre-Scan) und nach Behandlung (Tag 3) behandelt. (B) entsprechende quantitative Analyse von CLI und PET Ergebnisse (n = 3) und ihre Korrelationen. Bilder von (6) angepasst ist.arge.jpg &quot;target =&quot; _blank &quot;&gt; Klicken Sie hier für eine größere Abbildung zu sehen.

<ol>
	<li>Weissleder, R., Mahmood, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+imaging.">Molecular imaging.</a> Radiology. 219 (2), 316 (2001).</li><li>Chen, K., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Positron+emission+tomography+imaging+of+cancer+biology:+current+status+and+future+prospects.">Positron emission tomography imaging of cancer biology: current status and future prospects.</a> Semin. Oncol. 38 (1), 70 (2011).</li><li>Solomon, M., Liu, Y., Berezin, M. 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=Optical+imaging+in+cancer+research:+basic+principles,+tumor+detection,+and+therapeutic+monitoring.">Optical imaging in cancer research: basic principles, tumor detection, and therapeutic monitoring.</a> Med. Princ. Pract. 20 (5), 397 (2011).</li><li>Liu, H., Ren, G., Miao, Z., 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=Molecular+Optical+Imaging+with+Radioactive+Probes.">Molecular Optical Imaging with Radioactive Probes.</a> PLoS One. 5 (3), e9470 (2010).</li><li>Robertson, R., Germanos, M. S., Li, 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=Optical+imaging+of+Cerenkov+light+generation+from+positron-emitting+radiotracers.">Optical imaging of Cerenkov light generation from positron-emitting radiotracers.</a> Phys. Med. Biol. 54 (16), N355 (2009).</li><li>Xu, Y., Liu, H., Cheng, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Harnessing+the+power+of+radionuclides+for+optical+imaging:+Cerenkov+luminescence+imaging.">Harnessing the power of radionuclides for optical imaging: Cerenkov luminescence imaging.</a> J. Nucl. Med. 52 (12), 2009 (2011).</li><li>Cerenkov, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visible+emission+of+clean+liquids+by+action+of+g-radiation.">Visible emission of clean liquids by action of g-radiation.</a> Dokl Akad Nauk SSSR. 2, 451 (1934).</li><li>Cerenkov, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visible+radiation+produced+by+electrons+moving+in+a+medium+with+velocities+exceeding+that+of+light.">Visible radiation produced by electrons moving in a medium with velocities exceeding that of light.</a> Phys Rev. 52 (4), 0378 (1937).</li><li>Boschi, F., Calderan, L., D'Ambrosio, D., 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=In+vivo+18F-FDG+tumour+uptake+measurements+in+small+animals+using+Cerenkov+radiation.">In vivo 18F-FDG tumour uptake measurements in small animals using Cerenkov radiation.</a> Eur. J. Nucl. Med. Mol. Imaging. 38 (1), 120 (2011).</li><li>Liu, H., Ren, G., Liu, 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=Optical+imaging+of+reporter+gene+expression+using+a+positron-emission-tomography+probe.">Optical imaging of reporter gene expression using a positron-emission-tomography probe.</a> J. Biomed. Opt. 15 (6), 060505 (2010).</li><li>Park, J. C., Yu, M. K., An, G. 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=Facile+preparation+of+a+hybrid+nanoprobe+for+triple-modality+optical/PET/MR+imaging.">Facile preparation of a hybrid nanoprobe for triple-modality optical/PET/MR imaging.</a> Small. 6 (24), 2863 (2010).</li><li>Xu, Y., Chang, E., Liu, 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=Proof-of-concept+study+of+monitoring+cancer+drug+therapy+with+cerenkov+luminescence+imaging.">Proof-of-concept study of monitoring cancer drug therapy with cerenkov luminescence imaging.</a> J. Nucl. Med. 53 (2), 312 (2012).</li><li>Ellis, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bevacizumab.">Bevacizumab.</a> Nat. Rev. Drug Discov. , (2005).</li><li>Hochster, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bevacizumab+in+combination+with+chemotherapy:+first-line+treatment+of+patients+with+metastatic+colorectal+cancer.">Bevacizumab in combination with chemotherapy: first-line treatment of patients with metastatic colorectal cancer.</a> Semin. Oncol. 33, (2006).</li><li>Dothager, R. S., Goiffon, R. J., Jackson, E., 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=Cerenkov+radiation+energy+transfer+(CRET)+imaging:+a+novel+method+for+optical+imaging+of+PET+isotopes+in+biological+systems.">Cerenkov radiation energy transfer (CRET) imaging: a novel method for optical imaging of PET isotopes in biological systems.</a> PLoS One. 5 (10), e13300 (2010).</li><li>Hu, Z., Liang, J., Yang, W., 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=Experimental+Cerenkov+luminescence+tomography+of+the+mouse+model+with+SPECT+imaging+validation.">Experimental Cerenkov luminescence tomography of the mouse model with SPECT imaging validation.</a> Opt. Express. 18 (24), 24441 (2010).</li><li>Park, J. C., Il An, G., Park, S. 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=Luminescence+imaging+using+radionuclides:+a+potential+application+in+molecular+imaging.">Luminescence imaging using radionuclides: a potential application in molecular imaging.</a> Nucl. Med. Biol. 38 (3), 321 (2011).</li><li>Holland, J. P., Normand, G., Ruggiero, 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=Intraoperative+imaging+of+positron+emission+tomographic+radiotracers+using+Cerenkov+luminescence+emissions.">Intraoperative imaging of positron emission tomographic radiotracers using Cerenkov luminescence emissions.</a> Mol. Imaging. 10 (3), 177 (2011).</li><li>Liu, H., Carpenter, C. M., Jiang, 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=Intraoperative+imaging+of+tumors+using+Cerenkov+luminescence+endoscopy:+a+feasibility+experimental+study.">Intraoperative imaging of tumors using Cerenkov luminescence endoscopy: a feasibility experimental study.</a> J. Nucl. Med. , (2012).</li></ol>

Keine Interessenskonflikte erklärt.

CLI wird als vielversprechender molekulare Bildgebung Technik, Potenziale gefunden hat in vielen Grundlagenforschung Anwendungen und sogar den klinischen Einsatz 4,5,15,16,17 Schwellenländern. Die wichtigsten Vorteile von CLI über traditionelle nukleare Bildgebungsverfahren wie PET Stammzellen aus der Verwendung von OI Instrumente, die einfacher zu bedienen sind, durch kurze Messzeit und hoher Durchsatz gekennzeichnet, deutlich günstiger und besser verfügbar für Forscher. Darüber, was setzt CLI abgesehe...

<table border="1"&gt;<tr&gt;<td<strong&gt; Name</strong</td&gt;<td<strong&gt; Unternehmen</strong</td&gt;<td<strong&gt; Katalog-Nummer</strong</td&gt;</tr&gt;<tr&gt;<td&gt; H460 Cell Line</td&gt;<td&gt; American Type Culture Collection</td&gt;<td&gt; ATCC-Nummer: HTB-177</td&gt;</tr&gt;<tr&gt;<td&gt; RPMI 1640 Medium</td&gt;<td&gt; Invitrogen Life Technologies</td&gt;<td&gt; 12633-012</td&gt;</tr&gt;<tr&gt;<td&gt; Fetal Bovine Serum</td&gt;<td&gt; Invitrogen Life Technologies</td&gt;<td&gt; 10091-148</td&gt;</tr&gt;<tr&gt;<td&gt; Penicillin / Streptomycin</td&gt;<td&gt; Invitrogen Life Technologies</td&gt;<td&gt; 15640-055</td&gt;</tr&gt;<tr&gt;<td&gt; Phosphate-Buffered Saline</td&gt;<td&gt; Invitrogen Life Technologies</td&gt;<td&gt; 10010-023</td&gt;</tr&gt;<tr&gt;<td&gt; Female Nacktmausmodell</td&gt;<td&gt; Charles River Laboratories, Inc.</td&gt;<td&gt; Strain Code: 088</td&gt;</tr&gt;<tr&gt;<td&gt; Bevacizumab (Avastin)</td&gt;<td&gt; Genentech / Roche</td&gt;<td&gt; N / A</td&gt;</tr&gt;<tr&gt;<td&gt; MicroPET Rodent R4</td&gt;<td&gt; Siemens Medical Solutions USA, Inc.</td&gt;<td&gt; N / A</td&gt;</tr&gt;<tr&gt;<td&gt; Isofluran (Aerrane)</td&gt;<td&gt; Baxter</td&gt;<td&gt; Baxter-Nummer: AHN3637</td&gt;</tr&gt;<tr&gt;<td&gt; IVIS Spectrum</td&gt;<td&gt; Caliper Life Sciences</td&gt;<td&gt; N / A</td&gt;</tr&gt;</table

cerenkov luminescence imaging (cli) for cancer therapy monitoring

In der molekularen Bildgebung sind Positronen-Emissions-Tomographie (PET) und Abbildungsoptik (OI) zwei der wichtigsten und am häufigsten verwendeten somit Modalitäten 1-3. PET zeichnet sich durch seine hervorragende Empfindlichkeit und Quantifizierung Fähigkeit, während OI zeichnet für Nicht-Strahlung, relativ niedrige Kosten, kurze Scanzeit, hohen Durchsatz und breite Verfügbarkeit, bis zum Grundlagenforscher aus. Allerdings haben beide Modalitäten ihrer Mängel als gut. PET leidet schlechte räumliche Auflösung und hohen Kosten, während OI ist vor allem auf präklinischen Anwendungen aufgrund ihrer begrenzten Eindringen in das Gewebe zusammen mit prominenten Streuung optischer Signale durch die Dicke von lebendem Gewebe beschränkt. Kürzlich wurde eine Brücke zwischen PET und OI wurde mit der Entdeckung der Cerenkov Lumineszenz Imaging (CLI) 4-6 entstanden. CLI ist ein neues bildgebendes Verfahren, das Cerenkov Strahlung (CR) nutzt, um Bild Radionuklide mit OI Instrumente. Russische Nobel laureate Alexejewitsch Cerenkov und seine Kollegen ursprünglich entdeckt CR 1934. Es ist eine Form von elektromagnetischer Strahlung emittiert werden, wenn ein geladenes Teilchen bewegt sich mit einer Geschwindigkeit superluminalen in einem dielektrischen Medium 7,8. Das geladene Teilchen, ob Positronen oder Elektronenstrahlen, stört das elektromagnetische Feld des Mediums durch Verschieben der Elektronen in seiner Atome. Nach dem Passieren der Unterbrechung Photonen emittiert als die verdrängten Elektronen in den Grundzustand zurückzukehren. Zum Beispiel wurde ein 18 F Zerfall schätzungsweise durchschnittlich 3 Photonen in Wasser 5 herzustellen. Seit ihrer Entstehung hat CLI für seine Verwendung in einer Vielzahl von Anwendungen einschließlich präklinischen in vivo Tumordarstellung, Reportergens Bildgebung Radiotracers Entwicklung, multimodale Bildgebung unter anderem 4,5,9,10,11 untersucht. Der wichtigste Grund, warum CLI viel Erfolg genossen hat, so weit ist, dass diese neue Technologie zu Nutze macht die geringe Zusammenarbeitst und breite Verfügbarkeit von OI Bild Radionuklide, die nur durch teurere und weniger verfügbar nukleare Bildgebungsverfahren wie PET abgebildet werden verwendet. Hier präsentieren wir die Methode der Verwendung von CLI, um Krebs medikamentöse Therapie zu überwachen. Unsere Gruppe hat vor kurzem diese neue Anwendung untersucht und validiert die Machbarkeit von einem Proof-of-Concept-Studie 12. Wir haben gezeigt, dass CLI und PET ausgezeichnete Korrelationen zeigten in verschiedenen Tumorxenotransplantaten und Imaging-Sonden. Dies steht im Einklang mit dem übergeordneten Prinzip der CR, dass CLI wesentlichen visualisiert die gleichen Radionuklide als PET. Wir haben uns für Bevacizumab (Avastin, Genentech / Roche) als unser therapeutisches Mittel, weil es eine bekannte Angiogenese-Hemmer 13,14 ist. Reifung dieser Technologie in der nahen Zukunft kann sich vorstellen, einen signifikanten Einfluss auf die präklinischen Entwicklung, Screening sowie Therapie-Monitoring der Patienten, die Behandlungen haben werden.

Watch this Scientific Journal Video about Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring at JoVE.com

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

Verwendung von Cerenkov Lumineszenz Imaging (CLI) für die Überwachung der präklinischen Behandlung von Krebs wird hier beschrieben. Diese Methode nutzt die Vorteile der Cerenkov Strahlung (CR) und optische Bildgebung (OI) an radioaktiv markierten Sonden visualisieren und stellt somit eine Alternative zu PET in der präklinischen therapeutische Überwachung und Wirkstoff-Screening.

Cerenkov Lumineszenz Imaging (CLI) for Cancer Therapy Überwachung

In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

cerenkov-luminescence-imaging-cli-for-cancer-therapy-monitoring

Research

JoVE Journal

Medicine

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

13.6K Aufrufe.  Stanford University. Verwendung von Cerenkov Lumineszenz Imaging (CLI) für die Überwachung der präklinischen Behandlung von Krebs wird hier beschrieben. Diese Methode nutzt die Vorteile der Cerenkov Strahlung (CR) und optische Bildgebung (OI) an radioaktiv markierten Sonden visualisieren und stellt somit eine Alternative zu PET in der präklinischen therapeutische Überwachung und Wirkstoff-Screening.

Serie: Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence imaging using IVIS Spectrum was employed to monitor the distribution and development of the tumor cells following the delivery procedure including DLIT reconstruction to measure the tumor signal and its location.
Development of metastatic lesions to the bone tissues triggers osteolytic activity and lesions to tibia and femur were evaluated longitudinally using micro CT. Imaging was performed using a Quantum FX micro CT system with fast imaging and low X-ray dose. The low radiation dose allows multiple imaging sessions to be performed with a cumulative X-ray dosage far below LD50. A mouse imaging shuttle device was used to sequentially image the mice with both IVIS Spectrum and Quantum FX achieving accurate animal positioning in both the bioluminescence and CT images. The optical and CT data sets were co-registered in 3-dimentions using the Living Image 4.1 software. This multi-mode approach allows close monitoring of tumor growth and development simultaneously with osteolytic activity.

An experimental mouse model of bone metastasis was established following intracardiac delivery of luciferase expressing mammary tumor cells. Tumor development and resulted osteolytic lesion were monitored longitudinally with bioluminescence and micro CT imaging.

Überwachung Tumormetastasen und Osteolysen mit Biolumineszenz und Micro-CT Imaging

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

Forschung

Medizin

Modeling and Imaging 3-Dimensional Collective Cell Invasion

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy tissue is the root cause of disease and death. For other cancers (e.g. breast, lung, etc.), it is the process of metastasis, in which tumor cells move from a primary tumor mass, colonize distal sites and ultimately contribute to organ failure, that eventually leads to morbidity and mortality 3. It has been estimated that invasion and metastasis are responsible for 90&#37; of cancer deaths 4. As a result, there has been intense interest in identifying the molecular processes and critical protein mediators of invasion and metastasis for the purposes of improving diagnosis and treatment 5.
A challenge for cancer scientists is to develop invasion assays that sufficiently resemble the in vivo situation to enable accurate disease modeling 6. Two-dimensional cell motility assays are only informative about one aspect of invasion and do not take into account extracellular matrix (ECM) protein remodeling which is also a critical element. Recently, research has refined our understanding of tumor cell invasion and revealed that individual cells may move by elongated or rounded modes 7. In addition, there has been greater appreciation of the contribution of collective invasion, in which cells invade in strands, sheets and clusters, particularly in highly differentiated tumors that maintain epithelial characteristics, to the spread of cancer 8.
We present a refined method 9 for examining the contributions of candidate proteins to collective invasion 10. In particular, by engineering separate pools of cells to express different fluorescent proteins, it is possible to molecularly dissect the activities and proteins required in leading cells versus those required in following cells. The use of RNAi provides the molecular tool to experimentally disassemble the processes involved in individual cell invasion as well as in different positions of collective invasion. In this procedure, mixtures of fluorescently-labeled cells are plated on the bottom of a Transwell insert previously filled with Matrigel ECM protein, then allowed to invade "upwards" through the filter and into the Matrigel. Reconstruction of z-series image stacks, obtained by confocal imaging, into three-dimensional representations allows for visualization of collectively invading strands and analysis of the representation of fluorescently-labeled cells in leading versus following positions.

Models of tumor cell invasion into three-dimensional extracellular matrix better reflect the in vivo situation than two-dimensional motility assays. Using matrix invasion assays combined with confocal imaging of fluorescently-labeled cells, detailed information on invasion modes and the distinct contributions of leading versus following cells can be obtained.

Modeling and Imaging 3-Dimensional Collective Zellinvasion

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy ...

Protocols for Assessing Radiofrequency Interactions with Gold Nanoparticles and Biological Systems for Non-invasive Hyperthermia Cancer Therapy

Cancer therapies which are less toxic and invasive than their existing counterparts are highly desirable. The use of RF electric-fields that penetrate deep into the body, causing minimal toxicity, are currently being studied as a viable means of non-invasive cancer therapy. It is envisioned that the interactions of RF energy with internalized nanoparticles (NPs) can liberate heat which can then cause overheating (hyperthermia) of the cell, ultimately ending in cell necrosis.
In the case of non-biological systems, we present detailed protocols relating to quantifying the heat liberated by highly-concentrated NP colloids. For biological systems, in the case of in vitro experiments, we describe the techniques and conditions which must be adhered to in order to effectively expose cancer cells to RF energy without bulk media heating artifacts significantly obscuring the data. Finally, we give a detailed methodology for in vivo mouse models with ectopic hepatic cancer tumors.

We describe the protocols used to investigate the interactions of 13.56 MHz radiofrequency (RF) electric-fields with gold nanoparticle colloids in both non-biological and biological systems (in vitro/vivo). These interactions are being investigated for applications in cancer therapy.

Protokolle für die Beurteilung der Radiofrequenz-Wechselwirkungen mit Gold-Nanopartikel und biologische Systeme für die Nicht-invasive Krebstherapie Hyperthermie

Cancer therapies  which are less toxic and invasive than their existing counterparts are highly  desirable. The use of RF electric-fields that penetrate ...

Cerenkov Luminescence Imaging of Interscapular Brown Adipose Tissue

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely related to metabolism and energy expenditure, and its dysfunction is one important contributor for obesity and diabetes. Contrary to previous belief, recent PET/CT imaging studies indicated the BAT depots are still present in human adults. PET imaging clearly shows that BAT has considerably high uptake of 18F-FDG under certain conditions. In this video report, we demonstrate that Cerenkov luminescence imaging (CLI) with 18F-FDG can be used to optically image BAT in small animals. BAT activation is observed after intraperitoneal injection of norepinephrine (NE) and cold treatment, and depression of BAT is induced by long anesthesia. Using multiple-filter Cerenkov luminescence imaging, spectral unmixing and 3D imaging reconstruction are demonstrated. Our results suggest that CLI with 18F-FDG is a practical technique for imaging BAT in small animals, and this technique can be used as a cheap, fast, and alternative imaging tool for BAT research.

In this video report, we show the application of Cerenkov Luminescence Imaging (CLI) for interscapular brown adipose tissue in mice under activated and depressed conditions.

Cerenkov Lumineszenz-Imaging von interskapulären braunes Fettgewebe

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Mausmodell für nicht-invasive Bildgebungs zu erfassen und zu überwachen Eierstockkrebs Rezidiv

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Dynamic Lung Tumor Tracking for Stereotactic Ablative Body Radiation Therapy

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable risks for normal tissue injury and the hazards of physical tumor miss. A first-of-its-kind SBRT platform achieves high-precision ablative radiation treatment through a combination of versatile real-time imaging solutions and sophisticated tumor tracking capabilities. It uses dual-diagnostic kV x-ray units for stereoscopic open-loop feedback of cancer target intrafraction movement occurring as a consequence of respiratory motions and heartbeat. Image-guided feedback drives a gimbaled radiation accelerator (maximum 15 x 15 cm field size) capable of real-time &#177;4 cm pan-and-tilt action. Robot-driven &#177;60&#176; pivots of an integrated &#177;185&#176; rotational gantry allow for coplanar and non-coplanar accelerator beam set-up angles, ultimately permitting unique treatment degrees of freedom. State-of-the-art software aids real-time six dimensional positioning, ensuring irradiation of cancer targets with sub-millimeter accuracy (0.4 mm at isocenter). Use of these features enables treating physicians to steer radiation dose to cancer tumor targets while simultaneously reducing radiation dose to normal tissues. By adding respiration correlated computed tomography (CT) and 2-[18F] fluoro-2-deoxy-&#7429;-glucose (18F-FDG) positron emission tomography (PET) images into the planning system for enhanced tumor target contouring, the likelihood of physical tumor miss becomes substantially less1. In this article, we describe new radiation plans for the treatment of moving lung tumors.

Stereotactic ablative body radiation therapy (SBRT) involves the precise delivery of high-dose radiation to cancer tumor targets. A novel SBRT platform offers a first-of-its-kind gimbaled radiation accelerator mounted within a pivoting O-ring gantry capable of dynamic image-guided tumor tracking. Here, we describe dynamic tumor tracking for lung targets.

Dynamische Lungentumor Tracking für stereotaktische Strahlentherapie Körper Ablative

Physicians considering stereotactic ablative body radiation therapy (SBRT) for the treatment of extracranial cancer targets must be aware of the sizeable ...

An Organotypic High Throughput System for Characterization of Drug Sensitivity of Primary Multiple Myeloma Cells

In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and heterogeneity of patient-derived multiple myeloma (MM) cells. This approach consists in seeding primary MM cells freshly extracted from bone marrow aspirates into microfluidic chambers implemented in multi-well plates, each consisting of a reconstruction of the bone marrow microenvironment, including extracellular matrix (collagen or basement membrane matrix) and stroma (patient-derived mesenchymal stem cells) or human-derived endothelial cells (HUVECs). The chambers are drugged with different agents and concentrations, and are imaged sequentially for 96 hr through bright field microscopy, in a motorized microscope equipped with a digital camera. Digital image analysis software detects live and dead cells from presence or absence of membrane motion, and generates curves of change in viability as a function of drug concentration and exposure time. We use a computational model to determine the parameters of chemosensitivity of the tumor population to each drug, as well as the number of sub-populations present as a measure of tumor heterogeneity. These patient-tailored models can then be used to simulate therapeutic regimens and estimate clinical response.

We describe a high-throughput drug sensitivity assay for primary multiple myeloma cells. It consists of a reconstruction of the bone marrow microenvironment (including extracellular matrix and stroma) in multi-well plates, and a non-invasive method for longitudinal quantification of cell viability.

Ein organ High Throughput-System zur Charakterisierung von Arzneimittelempfindlichkeit von Primary Multiple Myelomzellen

In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and ...

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

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of stroke than relying on indirect measurements, and will be a potent and robust vascular research tool. We use an optical imaging approach that labels thrombi with a molecular imaging thrombus marker&#160;&#8212; a Cy5.5 near-infrared fluorescent (NIRF) probe that is covalently linked to the fibrin strands of the thrombus by the fibrin-crosslinking enzymatic action of activated coagulation factor XIIIa during the process of clot maturation. A micro-computed tomography (microCT)-based approach uses thrombus-seeking gold nanoparticles (AuNPs) functionalized to target the major component of the clot: fibrin. This paper describes a detailed protocol for the combined in vivo microCT and ex vivo NIRF imaging of thromboemboli in a mouse model of embolic stroke. We show that in vivo microCT and fibrin-targeted glycol-chitosan AuNPs (fib-GC-AuNPs) can be used for visualizing both in situ thrombi and cerebral embolic thrombi. We also describe the use of in vivo microCT-based direct thrombus imaging to serially monitor the therapeutic effects of tissue plasminogen activator-mediated thrombolysis. After the last imaging session, we demonstrate by ex vivo NIRF imaging the extent and the distribution of residual thromboemboli in the brain. Finally, we describe quantitative image analyses of microCT and NIRF imaging data. The combined technique of direct thrombus imaging allows two independent methods of thrombus visualization to be compared: the area of thrombus-related fluorescent signal on ex vivo NIRF imaging vs. the volume of hyperdense microCT thrombi in vivo.

This protocol describes the application of combined near-infrared fluorescent (NIRF) imaging and micro-computed tomography (microCT) for visualizing cerebral thromboemboli. This technique allows the quantification of thrombus burden and evolution. The NIRF imaging technique visualizes fluorescently labeled thrombus in excised brain, while the microCT technique visualizes thrombus inside living animals using gold-nanoparticles.

Kombinierte Near-Infrarot-Fluoreszenz-Bildgebung und Mikrocomputertomographie für Direkt Visualizing Cerebral Thromboembolie

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

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

Quantitative Visualisierung von Leukozyten-Infiltrat in einem Murin-Modell der Fulminant-Myokarditis durch Licht-Blatt-Mikroskopie

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Cardiac Magnetic Resonance for the Evaluation of Suspected Cardiac Thrombus: Conventional and Emerging Techniques

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The appearance of a mass on certain magnetic resonance (MR) sequences can help differentiate a thrombus from competing diagnoses such as a tumor. T1 and T2 signal characteristics of a thrombus are related to the evolution of hemoglobin properties. A thrombus typically does not enhance following contrast administration, which also helps differentiation from a tumor. We also highlight the emerging role of T1 mapping in the evaluation of a thrombus, which can add another level of support in diagnosis. Prior to any CMR exam, patient screening and interviews are critical to ensure safety and to optimize patient comfort. Effective communication during the exam between the technologist and the patient promotes proper breath holding technique and higher quality images. Volumetric post processing and structured reporting are helpful to ensure that the radiologist answers the ordering services&#39; question and communicates these results effectively. Optimal pre-MR safety evaluation, CMR exam execution, and post exam processing and reporting allow for delivery of high quality radiological service in the evaluation of a suspected cardiac thrombus.

The goal of this article is to describe how cardiac magnetic resonance can be used for the evaluation and diagnosis of a suspected cardiac thrombus. The method presented will describe data acquisition as well as the pre-procedure and post-procedure protocol.

Herz-Magnetresonanz zur Beurteilung von vermutetem Herzthrombus: Konventionelle und aufkommende Techniken

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The ...