window chamber surgery in tumor cell injected mice for tumor growth visualization and imaging

Veiledet tumorinokulering og multimodal avbildning hos mus

Tumor microvasculature is an important component of the tumor microenvironment that can be a target for therapy and a determinant of treatment response. In the preclinical setting, the microvasculature is typically studied using intravital microscopy in orthotopic or heterotopic window chamber animal models1,2. This has several advantages over histological studies since the imaging is done in live tissues and the tumor can be monitored longitudinally over several weeks or even months2,3. These studies can leverage the high-resolution imaging capabilities of intravital microscopy to study the delivery of therapeutics to the tumor4,5, the causes of treatment resistance6, and the response of the micro vessels to therapies such as antiangiogenic treatment7,8 and radiotherapy2,9.
Intravital microscopy clearly plays an important role in preclinical cancer research; however, how can tumor microenvironmental features be measured in the clinic? Microvascular information would be useful in the clinic for measuring blood supply and tumor cell hypoxia, which is important for determining treatment resistance in radiotherapy10, as well as the ability of the microvasculature to deliver chemotherapeutic agents to the surrounding tumor cells11. For example, in radiotherapy, spatial information on the structure and function of the tumor microvasculature may help personalize a patient&#39;s treatment plan by adjusting the fractionation schedule or by preferentially boosting the dose to avascular and likely hypoxic regions12.
Intravital microscopy can measure these important microvascular features since it has a very high resolution (&#956;m scale); however, its depth penetration into tissue is limited to several hundred microns or a few millimeters, at most making clinical implementation challenging. Indeed, there are some novel applications of intravital microscopy in the clinic13; however, these are still limited to examinations of near-surface level tissue such as the skin14 or mucosal/endothelial linings of various body cavities via flexible catheters/endoscopes15,16.
More commonly, the microvasculature is studied using imaging modalities such as CT17 or MRI18. These clinical imaging modalities can image to any depth within the body, but they have a much lower spatial resolution (mm scale). Thus, there is a need to bridge this resolution gap between preclinical intravital microscopy and clinical imaging modalities to bring high-resolution and detailed microvascular information into the clinic19. Several functional imaging methods have been developed to improve the microvascular imaging capabilities of clinical imaging modalities such as dynamic contrast-enhanced (DCE) MRI and CT20, and Intravoxel incoherent motion (IVIM) MRI21. However, these are model-based methods that provide indirect measurements of the microvasculature and thus, must be validated with appropriate &#34;ground truth&#34; measurements of the microvasculature19,22.
We have developed a dorsal skinfold window chamber (DSFC) tumor mouse model to bridge this gap between preclinical intravital microscopy and clinically applicable imaging modalities such as CT and MRI. The DSFC provides direct access to the tumor for high-resolution, intravital microscopy imaging through a glass window but also clinically applicable imaging such as MRI as it is made of MR-compatible materials (plastic and glass). Furthermore, an included MATLAB code performs multimodality 3D co-registration for direct spatial correlations between preclinical intravital microscopy and clinically applicable imaging modalities. Here we will describe the design and surgery to install the DSFC as well as the procedure to co-register intravital microscopy and clinically applicable imaging modalities.

Forfatter Spotlight: Integrering av høyoppløselig intravital avbildning og MR for å forbedre stereotaktisk kroppsstrålebehandlingsplanlegging

En dorsal hudfold vinduskammer svulstmusemodell for kombinert intravital mikroskopi og magnetisk resonansavbildning i translasjonell kreftforskning

Preclinical intravital imaging has very high resolution, but limited depth penetration in the tissue, which makes it very good for preclinical imaging studies. On the other hand, MRI is much more clinically applicable and has a higher depth penetration, but very low spatial resolution. The goal of this study is to correlate these two modalities with each other in order to better translate our findings from preclinical intravital microscopy into the clinic via magnetic resonance imaging.Perfusion-sensitive imaging methods in MRI and biophotonics, including our optical coherence tomography and geography system, are revealing how the microvasculature impacts tumor response to hypofractionated radiotherapy. 3D printing is also aiding through cost-effective custom tool manufacturing, facilitating longitudinal studies. Finally, AI image analysis is facilitating the identification of novel radiotherapy predictive biomarkers.We've made several findings concerning the tumor response to hypofractionated radiotherapy, including more recently, the temporal kinetics of both radiobiologically significant and abstracted AI-derived tissue response metrics. Additionally, we have demonstrated the promise for multimodal imaging being performed more accurately, correlating both MRI, macroscopic and optical coherence tomography, microscopic, angiography-derived metrics. By extending the effective length of longitudinal studies, we may better identify predictive biomarkers for long-term tumor treatment response.Additionally, by improving robustness of co-registration between macroscopic clinically available imaging modalities and microscopic preclinical imaging modalities, we may enhance the translatability of our findings from benchtop to bedside. During a stereotactic body radiation therapy, high doses of radiation are delivering a reduced number of fractions. One potential radiobiological target associated with increased cell death in SVRT is the tumor microvasculature.By combining high-resolution optical angiography and MRI, we want to enable functional MRI microvascular imaging for patient-specific SVRT treatment planning and improved outcomes.

Vinduskammerkirurgi i tumorcelleinjiserte mus for visualisering og bildebehandling av tumorvekst

For å begynne, steriliser hovedvinduskammerdelene i et flytende desinfeksjonsmiddel på høyt nivå i 12 minutter. Etterfulgt av bløtlegging i isopropylalkohol i fem minutter. Administrer ett milligram per kilo buprenorfin med forsinket frigjøring til den tumorcelleinjiserte musen en til tre timer før operasjonen.For desinfisering, skrubb den bedøvede og barberte musen med 7,5% povidon-jodoppløsning. Fjern den desinfiserende skrubben med 70% isopropylalkohol. Påfør en 10% povidon-jod-løsning på huden.Påfør til slutt veterinær øyesmøremiddel for å forhindre tørrhet. Løft musens hud langs ryggraden og finn svulsten som vokser på en av de laterale hudfoldene. Plasser den bakre rammen til det dorsale hudfold-vinduskammeret, eller DSFC, slik at svulsten er sentrert innenfor rammen.Legg til tre suturer til de tre øverste hullene i DSFC og den kirurgiske guiden for å fikse posisjonen til bakrammen. Sett tre nåler gjennom bakrammen på DSFC inn i hullene som inneholder avstandsstykker. Bruk en kirurgisk markør til å markere tre punkter for DSFC-bakrammeavstandsstykkene på huden.Tegn en sirkel med en centimeter i diameter for å indikere hudområdet som skal fjernes. Etter å ha merket tre ekstra prikker, klipp suturene for å fjerne nålene og bakrammen. Fjern huden fra den markerte sirkelen ved hjelp av den kirurgiske mikrosaksen.Plasser deretter den midlertidige suturen for å feste vinduskammerdelene til huden. Stram enheten med en midlertidig sutur og sett frontrammen under huden gjennom hullet. Plasser permanente suturer gjennom hullene rundt omkretsen av de fremre og bakre rammene til DSFC.Etter å ha fjernet den midlertidige suturen, fest støtteklemmen til DSFC-bakrammen. Administrer fem milligram meloksikam per kilo kroppsvekt subkutant til musen for smerte og betennelsesreduksjon.

window-chamber-surgery-tumor-cell-injected-mice-for-tumor-growth

Research

JoVE Journal

Cancer Research

Window Chamber Surgery in Tumor Cell Injected Mice for Tumor Growth Visualization and Imaging

A Dorsal Skinfold Window Chamber Tumor Mouse Model for Combined Intravital Microscopy and Magnetic Resonance Imaging in Translational Cancer Research

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the ...