<meta charset="utf8"></head><body>氧化铁纳米颗粒用于小鼠胶质母细胞瘤的图像引导治疗递送

<ol>
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Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temozolomide+resistance+in+glioblastoma+multiforme.">Temozolomide resistance in glioblastoma multiforme.</a> Genes Dis. 3 (3), 198-210 (2016).</li><li>Dong, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+strategies+for+brain+drug+delivery.">Current strategies for brain drug delivery.</a> Theranostics. 8 (6), 1481-1493 (2018).</li><li>Teleanu, D. M., Chircov, C., Grumezescu, A. M., Volceanov, A., Teleanu, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood-brain+delivery+methods+using+nanotechnology.">Blood-brain delivery methods using nanotechnology.</a> Pharmaceutics. 10 (4), 268 (2018).</li><li>Ohta, 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=Investigating+the+optimum+size+of+nanoparticles+for+their+delivery+into+the+brain+assisted+by+focused+ultrasound-induced+blood-brain+barrier+opening.">Investigating the optimum size of nanoparticles for their delivery into the brain assisted by focused ultrasound-induced blood-brain barrier opening.</a> Sci Rep. 10 (1), 18220 (2020).</li><li>Teplyuk, N. 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=Therapeutic+potential+of+targeting+microrna-10b+in+established+intracranial+glioblastoma:+First+steps+toward+the+clinic.">Therapeutic potential of targeting microrna-10b in established intracranial glioblastoma: First steps toward the clinic.</a> EMBO Mol Med. 8 (3), 268-287 (2016).</li><li>Dube, T., Kumar, N., Bishnoi, M., Panda, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual+blood-brain+barrier-glioma+targeting+peptide-poly(levodopamine)+hybrid+nanoplatforms+as+potential+near+infrared+phototheranostic+agents+in+glioblastoma.">Dual blood-brain barrier-glioma targeting peptide-poly(levodopamine) hybrid nanoplatforms as potential near infrared phototheranostic agents in glioblastoma.</a> Bioconjug Chem. 32 (9), 2014-2031 (2021).</li><li>Yoo, 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=Design+of+nanodrugs+for+mirna+targeting+in+tumor+cells.">Design of nanodrugs for mirna targeting in tumor cells.</a> J Biomed Nanotechnol. 10 (6), 1114-1122 (2014).</li><li>Medarova, Z., Pham, W., Farrar, C., Petkova, V., Moore, A. <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+imaging+of+sirna+delivery+and+silencing+in+tumors.">In vivo imaging of sirna delivery and silencing in tumors.</a> Nature Medicine. 13 (3), 372-377 (2007).</li><li>Chen, 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=Co-administration+of+temozolomide+(tmz)+and+the+experimental+therapeutic+targeting+mir-10b,+profoundly+affects+the+tumorigenic+phenotype+of+human+glioblastoma+cells.">Co-administration of temozolomide (tmz) and the experimental therapeutic targeting mir-10b, profoundly affects the tumorigenic phenotype of human glioblastoma cells.</a> Front Mol Biosci. 10, 1179343 (2023).</li><li>Irtenkauf, S. 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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=Iron+oxide+nanoparticles:+Diagnostic,+therapeutic+and+theranostic+applications.">Iron oxide nanoparticles: Diagnostic, therapeutic and theranostic applications.</a> Adv Drug Deliv Rev. 138, 302-325 (2019).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Athymic nude &#34;J:NU&#34; mice</td>
			<td>Jackson Laboratory</td>
			<td>RRID:IMSR_JAX:007850</td>
			<td>Immunocompromised mouse model</td>
		</tr>
		<tr>
			<td>0.25% Trypsin</td>
			<td>Gibco</td>
			<td>25200-056</td>
			<td>Cell culture reagent for U251</td>
		</tr>
		<tr>
			<td>1.7 mL microcentrifuge tube</td>
			<td>DOT Scientific</td>
			<td>RN1700-GMT</td>
			<td>For tissue collection</td>
		</tr>
		<tr>
			<td>10 &#181;L, Neuros Syringe, Model 1701 RN, 33 gauge, Point Style 4</td>
			<td>Hamilton</td>
			<td>65460-06</td>
			<td>Syringe for intracranial implantation of tumor cells</td>
		</tr>
		<tr>
			<td>3M Vetbond</td>
			<td>3M</td>
			<td>1469SB</td>
			<td>Tissue adhesive for surgical site closure</td>
		</tr>
		<tr>
			<td>4% Paraformaldehyde&#160;</td>
			<td>Thermo Scientific</td>
			<td>J199943-K2</td>
			<td>Tissue fixing solution</td>
		</tr>
		<tr>
			<td>70% isopropoyl alcohol wipe</td>
			<td>Cardinal</td>
			<td>MW-APL</td>
			<td>Topical antiseptic wipe for tumor implantation and tail vein injection</td>
		</tr>
		<tr>
			<td>Aperio Versa</td>
			<td>Leica</td>
			<td></td>
			<td>For scanning of stained tissue section slides</td>
		</tr>
		<tr>
			<td>Betadine Surgical Scrub</td>
			<td>Purdue</td>
			<td>6761815101</td>
			<td>Topical antiseptic for tumor implantation</td>
		</tr>
		<tr>
			<td>BioSpec 70/30</td>
			<td>Bruker</td>
			<td></td>
			<td>Magnetic resonance imaging scanner</td>
		</tr>
		<tr>
			<td>Bone Wax</td>
			<td>Medline</td>
			<td>DYNJBW25</td>
			<td>Bone wax for sealing implantation site</td>
		</tr>
		<tr>
			<td>Burrs for Micro drill</td>
			<td>F.S.T.</td>
			<td>19007-05</td>
			<td>Drill burr used to make hole in skull for tumor implantation</td>
		</tr>
		<tr>
			<td>DAPI Fluoromount-G</td>
			<td>SouthernBiotech</td>
			<td>0100-20</td>
			<td>Tissue mounting media containing DAPI stain</td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s Modified Eagle Medium (DMEM)</td>
			<td>Gibco</td>
			<td>11995-065</td>
			<td>Cell culture media for U251</td>
		</tr>
		<tr>
			<td>Extra Fine Graefe Forceps</td>
			<td>F.S.T.</td>
			<td>11150-10</td>
			<td>Sugical tool for tumor implantation</td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Corning</td>
			<td>35-010-CV</td>
			<td>Cell culture media supplement for U251</td>
		</tr>
		<tr>
			<td>Fine Scissors - Sharp 10.5cm</td>
			<td>F.S.T.</td>
			<td>14060-10</td>
			<td>Sugical tool for tumor implantation</td>
		</tr>
		<tr>
			<td>Glydo (Lidocaine)</td>
			<td>Sagent</td>
			<td>673-76</td>
			<td>Topical analgesic for surgical site</td>
		</tr>
		<tr>
			<td>Ideal Micro Drill</td>
			<td>CellPoint Scientific</td>
			<td>67-1200A</td>
			<td>Drill used to make hole in skull for tumor implantation</td>
		</tr>
		<tr>
			<td>Insulin syringe 1CC 29G X 1/2&#34;</td>
			<td>Becton, Dickinson</td>
			<td>324704</td>
			<td>Syringe for D-Luciferin injection and tail vein injection of nanoparticles</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Covetrus</td>
			<td>11695067772</td>
			<td>Anethesia</td>
		</tr>
		<tr>
			<td>Isoflurane vaporizer</td>
			<td>SOMNI Scientific</td>
			<td>VS6002</td>
			<td>Anethesia apparatus</td>
		</tr>
		<tr>
			<td>IVIS SpectrumCT In Vivo Imaging System</td>
			<td>PerkinElmer/Revvity</td>
			<td>128201</td>
			<td>Bioluminescence and fluorescence imaging scanner</td>
		</tr>
		<tr>
			<td>IVISbrite D-Luciferin Potassium Salt</td>
			<td>PerkinElmer/Revvity</td>
			<td>122799-100MG</td>
			<td>Substrate for bioluminescence imaging</td>
		</tr>
		<tr>
			<td>Ketaset (Ketamine)</td>
			<td>Zoetis</td>
			<td>10004027</td>
			<td>Anesthetic for tumor implantation surgery</td>
		</tr>
		<tr>
			<td>Ketofen (Ketoprofen)</td>
			<td>Zoetis</td>
			<td>10004031</td>
			<td>Analgesic for tumor implantation surgery</td>
		</tr>
		<tr>
			<td>Leica CM1950</td>
			<td>Leica</td>
			<td>CM1950</td>
			<td>For cryosectioning of OCT-embedded samples</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Gibco</td>
			<td>14190-144</td>
			<td>Cell culture reagent and cell suspension solution for implantation of U251</td>
		</tr>
		<tr>
			<td>Penicillin-streptomycin</td>
			<td>Gibco</td>
			<td>15140-122</td>
			<td>Antibiotic for cell culture media for U251</td>
		</tr>
		<tr>
			<td>Puralube vet ointment</td>
			<td>MWI Veterinary</td>
			<td>27505</td>
			<td>Opthalmic eye ointment for protection during tumor implantation</td>
		</tr>
		<tr>
			<td>Ruler</td>
			<td>F.S.T.</td>
			<td>18000-30</td>
			<td>Used to measure drill site for implanation</td>
		</tr>
		<tr>
			<td>Tissue-Tek Cryomold&#160; Intermediate 15 x 15 x 5 mm</td>
			<td>Sakura</td>
			<td>4566</td>
			<td>Collection mold for collecting tissue samples</td>
		</tr>
		<tr>
			<td>Tissue-Tek&#160;O.C.T. Compound</td>
			<td>Sakura</td>
			<td>4583</td>
			<td>Freezing compound for collecting tissue samples</td>
		</tr>
		<tr>
			<td>U-251 MG cell line human</td>
			<td>Millapore Sigma</td>
			<td>9063001</td>
			<td>Human glioblastoma cell line</td>
		</tr>
		<tr>
			<td>Xylazine Injectable Solution, 100 mg/ml</td>
			<td>Covetrus</td>
			<td>1XYL006</td>
			<td>Paralytic for tumor implantation surgery</td>
		</tr>
	</tbody>
</table>

nanoparticle delivery of an oligonucleotide payload in a glioblastoma multiforme animal model

<meta charset="utf8"></head><body>作者聚焦：使用氧化铁纳米颗粒治疗胶质母细胞瘤的创新癌症疗法

多形性胶质母细胞瘤动物模型中寡核苷酸有效载荷的纳米颗粒递送

Glioblastoma multiforme (GBM) is the most common and aggressive form of primary brain malignancy for which there is no cure. The blood-brain barrier is a ...

nanoparticle-delivery-an-oligonucleotide-payload-glioblastoma

Without Section

Research

Nanoparticle Delivery of an Oligonucleotide Payload in a Glioblastoma Multiforme Animal Model

JoVE Journal

Cancer Research

1.1K 次观看.  Michigan State University. 我们的研究重点是使用我们的氧化铁纳米颗粒递送平台开发癌症疗法。我们的纳米平台的优势在于它能够穿过血脑屏障并在肿瘤中积累，从而提供治疗。目标是增加胶质母细胞瘤的可用疗法的小型武器库。血脑屏障对向胶质母细胞瘤提供治疗药物提出了挑战。许多化合物无法穿过屏障，因此几乎不可能全身施用用于治疗胶质母细胞瘤的药物。目...

视频: 作者聚焦：使用氧化铁纳米颗粒治疗胶质母细胞瘤的创新癌症疗法

Coculture Assays to Study Macrophage and Microglia Stimulation of Glioblastoma Invasion

Glioblastoma multiforme (grade IV glioma) is a very aggressive human cancer with a median survival of 1 year post diagnosis. Despite the increased understanding of the molecular events that give rise to glioblastomas, this cancer still remains highly refractory to conventional treatment. Surgical resection of high grade brain tumors is rarely complete due to the highly infiltrative nature of glioblastoma cells. Therapeutic approaches which attenuate glioblastoma cell invasion therefore is an attractive option. Our laboratory and others have shown that tumor associated macrophages and microglia (resident brain macrophages) strongly stimulate glioblastoma invasion. The protocol described in this paper is used to model glioblastoma-macrophage/microglia interaction using in vitro culture assays. This approach can greatly facilitate the development and/or discovery of drugs that disrupt the communication with the macrophages that enables this malignant behavior. We have established two robust coculture invasion assays where microglia/macrophages stimulate glioma cell invasion by 5 - 10 fold. Glioblastoma cells labelled with a fluorescent marker or constitutively expressing a fluorescent protein are plated without and with macrophages/microglia on matrix-coated polycarbonate chamber inserts or embedded in a three dimensional matrix. Cell invasion is assessed by using fluorescent microscopy to image and count only invasive cells on the underside of the filter. Using these assays, several pharmacological inhibitors (JNJ-28312141, PLX3397, Gefitinib, and Semapimod), have been identified which block macrophage/microglia stimulated glioblastoma invasion.

Understanding the malignant behavior of cancer requires creating accurate models of how tumor cells interact with components of the tumor microenvironment, such as macrophages. Here we describe two methods to study glioblastoma cell interaction with tumor associated macrophages and microglia where the effect on glioblastoma invasion is assessed.

共培养化验，以研究胶质母细胞瘤入侵的巨噬细胞和小胶质细胞刺激

Glioblastoma multiforme (grade IV glioma) is a very aggressive human cancer with a median survival of 1 year post diagnosis. Despite the increased ...

科研

癌症研究

PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without the ability to target a specific or well-delineated tumor volume. The delivery of radiation was achieved using fixed radiation sources or linear accelerators producing megavoltage (MV) X-rays. These devices are unable to achieve sub-millimeter precision required for small animals. Furthermore, the high doses delivered to healthy surrounding tissue hamper response assessment. To increase the translation between small animal studies and humans, our goal was to mimic the treatment of human glioblastoma in a rat model. To enable a more accurate irradiation in a preclinical setting, recently, precision image-guided small animal radiation research platforms were developed. Similar to human planning systems, treatment planning on these micro-irradiators is based on computed tomography (CT). However, low soft-tissue contrast on CT makes it very challenging to localize targets in certain tissues, such as the brain. Therefore, incorporating magnetic resonance imaging (MRI), which has excellent soft-tissue contrast compared to CT, would enable a more precise delineation of the target for irradiation. In the last decade also biological imaging techniques, such as positron emission tomography (PET) gained interest for radiation therapy treatment guidance. PET enables the visualization of e.g., glucose consumption, amino-acid transport, or hypoxia, present in the tumor. Targeting those highly proliferative or radio-resistant parts of the tumor with a higher dose could give a survival benefit. This hypothesis led to the introduction of the biological tumor volume (BTV), besides the conventional gross target volume (GTV), clinical target volume (CTV), and planned target volume (PTV).
At the preclinical imaging lab of Ghent University, a micro-irradiator, a small animal PET, and a 7 T small animal MRI are available. The goal was to incorporate MRI-guided irradiation and PET-guided sub-volume boosting in a glioblastoma rat model.

In the past, small animal irradiation was usually performed without the ability to target a well-delineated tumor volume. The goal was to mimic the treatment of human glioblastoma in rats. Using a small animal irradiation platform, we performed MRI-guided 3D conformal irradiation with PET-based sub-volume boosting in a preclinical setting.

PET 与 MRI 引导下微器对大鼠胶质瘤模型的照射

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without ...

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting immunodeficient mice subcutaneously with cancer cells and measuring the tumor mass after it becomes visible and palpable. Orthotopic injections of cancer cells aim to introduce the xenograft in the microenvironment that most closely resembles the tissue of origin of the tumor being studied. Brain cancer research requires intracranial injection of cancer cells to allow the tumor formation and analysis in the unique microenvironment of the brain. The in vivo imaging of intracranial xenografts monitors instantaneously the tumor mass of orthotopically engrafted mice. Here we report the use of fluorescence molecular tomography (FMT) of brain tumor xenografts. The cancer cells are first transduced with near infrared fluorescent proteins and then injected in the brain of immunocompromised mice. The animals are then scanned to obtain quantitative information about the tumor mass over an extended period of time. Cell pre-labeling allows for cost effective, reproducible, and reliable quantification of the tumor burden within each mouse. We eliminated the need for injecting imaging substrates, and thus reduced the stress on the animals. A limitation of this approach is represented by the inability to detect very small masses; however, it has better resolution for larger masses than other techniques. It can be applied to evaluate the efficacy of a drug treatment or genetic alterations of glioma cell lines and patient-derived samples.

Fluorescence Molecular Tomography for In Vivo Imaging of Glioblastoma Xenografts

Orthotopic intracranial injection of tumor cells has been used in cancer research to study brain tumor biology, progression, evolution, and therapeutic response. Here we present fluorescence molecular tomography of tumor xenografts, which provides real-time intravital imaging and quantification of a tumor mass in preclinical glioblastoma models.

胶质瘤移植中体内成像的荧光分子层析成像研究

Tumorigenicity is the capability of cancer cells to form a tumor mass. A widely used approach to determine if the cells are tumorigenic is by injecting ...

Predicting In Vivo Payloads Delivery using a Blood-brain Tumor-barrier in a Dish

Highly selective by nature, the blood-brain barrier (BBB) is essential for the brain homeostasis in physiological conditions. However, in the context of brain tumors, the molecular selectivity of BBB also shields the neoplastic cells by blocking the delivery of peripherally administered chemotherapies. The development of novel drugs (including nanoparticles) targeting malignant brain tumors ideally requires the use of preclinical animal models to study the drug&#8217;s transcytosis and antitumor efficacy. In order to comply with the 3R principle (refine, reduce, and replace) to reduce the number of laboratory animals in experimental setup and perform the high-throughput screening of a large library of antitumor agents, we developed a reproducible in vitro human and murine mimic of the blood-brain tumor-barrier (BBTB) using three-layered cultures of endothelial cells, astrocytes, and patient-derived glioblastoma spheres. For higher scalability and reproducibility, commercial cell lines or immortalized cells have been used in tailored conditions to allow the formation of a barrier resembling the actual BBB. Here we describe a protocol to obtain a BBTB mimic by culturing endothelial cells in contact with astrocytes at specific cell densities on inserts. This BBTB mimic can be used, for instance, for the quantification and confocal imaging of the nanoparticle passage through the endothelial and astrocytic barriers, in addition to the evaluation of the tumor cell targeting within the same assay. Moreover, we show that the obtained data can be used to predict the behavior of nanoparticles in preclinical animal models. In a broader perspective, this in vitro model could be adapted to other neurodegenerative diseases for the determination of the passage of new therapeutic molecules through the BBB and/or be supplemented with brain organoids to directly evaluate the efficacy of drugs.

Drug targeting to central nervous system tumors is a major challenge. Here we describe a protocol to produce an in vitro mimic of the blood-brain tumor-barrier using murine and/or human cells and discuss their relevance for the predictability of central nervous system tumor targeting in vivo.

利用血脑肿瘤屏障在盘中进行活体有效载荷传递预测

Highly selective by nature, the blood-brain barrier (BBB) is essential for the brain homeostasis in physiological conditions. However, in the context of ...

A 3D Spheroid Model for Glioblastoma

Two-dimensional (2D) cell cultures do not mimic in vivo tumor growth satisfactorily. Therefore, three-dimensional (3D) culture spheroid models were developed. These models may be particularly important in the field of neuro-oncology. Indeed, brain tumors have the tendency to invade the healthy brain environment. We describe herein an ideal 3D glioblastoma spheroid-based assay that we developed to study tumor invasion. We provide all technical details and analytical tools to successfully perform this assay.

Here, we describe an easy-to-use invasion assay for glioblastoma. This assay is suitable for glioblastoma stem-like cells. A Fiji macro for easy quantification of invasion, migration, and proliferation is also described.

胶质母细胞瘤的3D球形模型

Two-dimensional (2D) cell cultures do not mimic in vivo tumor growth satisfactorily. Therefore, three-dimensional (3D) culture spheroid models were ...

Positron Emission Tomography-based Dose Painting Radiation Therapy in a Glioblastoma Rat Model using the Small Animal Radiation Research Platform

A rat glioblastoma model to mimic chemo-radiation treatment of human glioblastoma in the clinic was previously established. Similar to the clinical treatment, computed tomography (CT) and magnetic resonance imaging (MRI) were combined during the treatment-planning process. Positron emission tomography (PET) imaging was subsequently added to implement sub-volume boosting using a micro-irradiation system. However, combining three imaging modalities (CT, MRI, and PET) using a micro-irradiation system proved to be labor-intensive because multimodal imaging, treatment planning, and dose delivery have to be completed sequentially in the preclinical setting. This also results in a workflow that is more prone to human error. Therefore, a user-friendly algorithm to further optimize preclinical multimodal imaging-based radiation treatment planning was implemented. This software tool was used to evaluate the accuracy and efficiency of dose painting radiation therapy with micro-irradiation by using an in silico study design. The new methodology for dose painting radiation therapy is superior to the previously described method in terms of accuracy, time efficiency, and intra- and inter-user variability. It is also an important step towards the implementation of inverse treatment planning on micro-irradiators, where forward planning is still commonly used, in contrast to clinical systems.

Here we present a protocol to perform preclinical positron emission tomography-based radiotherapy in a rat glioblastoma model using algorithms developed in-house to optimize the accuracy and efficiency.

使用小动物辐射研究平台在胶质母细胞瘤大鼠模型中基于正电子发射断层扫描的剂量绘画放射治疗

A rat glioblastoma model to mimic chemo-radiation treatment of human glioblastoma in the clinic was previously established. Similar to the clinical ...

Translational Orthotopic Models of Glioblastoma Multiforme

Genetically engineered mouse (GEM) models for human glioblastoma multiforme (GBM) are critical to understanding the development and progression of brain tumors. Unlike xenograft tumors, in GEMs, tumors arise in the native microenvironment in an immunocompetent mouse. However, the use of GBM GEMs in preclinical treatment studies is challenging due to long tumor latencies, heterogeneity in neoplasm frequency, and the timing of advanced grade tumor development. Mice induced via intracranial orthotopic injection are more tractable for preclinical studies, and retain features of the GEM tumors. We generated an orthotopic brain tumor model derived from a GEM model with Rb, Kras, and p53 aberrations (TRP), which develops GBM tumors&#160;displaying linear foci of necrosis by neoplastic cells, and dense vascularization analogous to human GBM. Cells derived from GEM GBM tumors are injected intracranially into wild-type, strain-matched recipient mice and reproduce grade IV tumors, therefore bypassing the long tumor latency period in GEM mice and allowing for the creation of large and reproducible cohorts for preclinical studies. The highly proliferative, invasive, and vascular features of the TRP GEM model for GBM are recapitulated in the orthotopic tumors, and histopathology markers reflect human GBM subgroups. Tumor growth is monitored by serial MRI scans. Due to the invasive nature of the intracranial tumors in immunocompetent models, carefully following the injection procedure outlined here is essential to prevent extracranial tumor growth.

Here, we describe a preclinical orthotopic mouse model for GBM, established by intracranial injection of cells derived from genetically engineered mouse model tumors. This model displays the disease hallmarks of human GBM. For translational studies, the mouse brain tumor is tracked by in vivo MRI and histopathology.

多形性胶质母细胞瘤的转化原位模型

Genetically engineered mouse (GEM) models for human glioblastoma multiforme (GBM) are critical to understanding the development and progression of brain ...

Glioblastoma Relapse Post-Resection Model for Therapeutic Hydrogel Investigations

Tumor recurrence is an important factor indicative of a poor prognosis in glioblastoma (GBM). Many studies are attempting to identify effective therapeutic strategies to prevent the recurrence of GBM after surgery. Bioresponsive therapeutic hydrogels capable of sustaining locally released drugs are frequently used for the local treatment of GBM after surgery. However, research is limited due to the lack of a suitable GBM relapse post-resection model. Here, a GBM relapse post-resection model was developed and applied in therapeutic hydrogel investigations. This model was constructed based on the orthotopic intracranial GBM model, which is widely used in studies on GBM. Subtotal resection was performed on the orthotopic intracranial GBM model mouse to mimic the clinical treatment. The residual tumor was used to indicate the size of the tumor growth. This model is easy to build, can better mimic the situation of GBM surgical resection, and can be applied in various studies on the local treatment of GBM relapse post-resection. As a result, the GBM relapse post-resection model provides a unique GBM recurrence model for effective local treatment studies of relapse post-resection.

The present protocol establishes a glioblastoma (GBM) relapse post-resection model using microscopy to investigate the therapeutic effect of an injectable, bioresponsive hydrogel in vivo.

用于治疗性水凝胶研究的胶质母细胞瘤复发术后切除模型

Tumor recurrence is an important factor indicative of a poor prognosis in glioblastoma (GBM). Many studies are attempting to identify effective ...

在小鼠中使用 2P-IVM 实时跟踪脑肿瘤中的纳米颗粒

Two-Photon Intravital Microscopy of Glioblastoma in a Murine Model

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the accumulation and distribution of macromolecules in brain tumors in vivo would greatly enhance our ability to understand and optimize drug delivery in preclinical models. This protocol describes a method for real-time in vivo tracking of intravenously administered fluorescent-labeled nanoparticles with two-photon intravital microscopy (2P-IVM) in a mouse model of glioblastoma (GBM).
The protocol contains a multi-step description of the procedure, including anesthesia and analgesia of experimental animals, creating a cranial window, GBM cell implantation, placing a head bar, conducting 2P-IVM studies, and post-surgical care for long-term follow-up studies. We show representative 2P-IVM imaging sessions and image analysis, examine the advantages and disadvantages of this technology, and discuss potential applications.
This method can be easily modified and adapted for different research questions in the field of in vivo preclinical brain imaging.

作者聚焦：优化胶质母细胞瘤治疗中药物递送和早期检测的多模式成像策略

We present a novel approach for two-photon microscopy of the tumor delivery of fluorescent-labeled iron oxide nanoparticles to glioblastoma in a mouse model.

小鼠模型中胶质母细胞瘤的双光子活体显微镜

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the ...