Inter and intra-tumor somatostatin receptor 2 heterogeneity influences peptide receptor radionuclide therapy response

Patients with neuroendocrine tumors (NETs) can be treated with peptide receptor radionuclide therapy (PRRT). Here, the somatostatin analogue octreotate radiolabeled with lutetium-177 is targeted to NET cells by binding to the somatostatin receptor subtype 2 (SST2). During radioactive decay, DNA damage is induced, leading to NET cell death. Although the therapy proves to be effective, mortality rates remain high. To appropriately select more optimal treatment strategies, it is essential to first better understand the radiobiological responses of tumor cells to PRRT. Methods: We analyzed PRRT induced radiobiological responses in SST2 expressing cells and xenografted mice using SPECT/MRI scanning and histological and molecular analyses. We measured [177Lu]Lu-DOTA-TATE uptake and performed analyses to visualize induction of DNA damage, cell death and other cellular characteristics. Results: The highest accumulation of radioactivity was measured in the tumor and kidneys. PRRT induced DNA damage signaling and repair in a time-dependent manner. We observed intra-tumor heterogeneity of DNA damage and apoptosis, which was not attributed to proliferation or bioavailability. We found a strong correlation between high DNA damage levels and high SST2 expression. PRRT elicited a different therapeutic response between models with different SST2 expression levels. Heterogeneous SST2 expression levels were also confirmed in patient NETs. Conclusion: Hete

Multi-modal image registration: matching MRI with histology

Spatial correspondence between histology and multi sequence MRI can provide information about the capabilities of non-invasive imaging to characterize cancerous tissue. However, shrinkage and deformation occurring during the excision of the tumor and the histological processing complicate the co registration of MR images with histological sections. This work proposes a methodology to establish a detailed 3D relation between histology sections and in vivo MRI tumor data. The key features of the methodology are a very dense histological sampling (up to 100 histology slices per tumor), mutual information based non-rigid B-spline registration, the utilization of the whole 3D data sets, and the exploitation of an intermediate ex vivo MRI. In this proof of concept paper, the methodology was applied to one tumor. We found that, after registration, the visual alignment of tumor borders and internal structures was fairly accurate. Utilizing the intermediate ex vivo MRI, it was possible to account for changes caused by the excision of the tumor: we observed a tumor expansion of 20%. Also the effects of fixation, dehydration and histological sectioning could be determined: 26% shrinkage of the tumor was found. The annotation of viable tissue, performed in histology and transformed to the in vivo MRI, matched clearly with high intensity regions in MRI. With this methodology, histological annotation can be directly related to the corresponding in vivo MRI. This is a vital step for the evaluation of the feasibility of multi-spectral MRI to depict histological ground-truth

Does the proteasome inhibitor bortezomib sensitize to DNA-damaging therapy in gastroenteropancreatic neuroendocrine neoplasms? – A preclinical assessment in vitro and in vivo

Background: Well-differentiated gastroenteropancreatic neuroendocrine neoplasms are rare tumors with a slow proliferation. They are virtually resistant to many DNA-damaging therapeutic approaches, such as chemo- and external beam therapy, which might be overcome by DNA damage inhibition induced by proteasome inhibitors suc

Development and in vivo validation of an MR-compatible temperature controllable superficial hyperthermia applicator for small animal studies

In this work, a superficial hyperthermia applicator was designed to be used in small animal studies. The applicator operates at 2.45 GHz and is compatible with a magnetic resonance scanner. A temperature controller was interfaced with the applicator to control the rate of change in temperature and the target temperature. Operation and robustness were validated through phantom and in vivo experiments

Development and in vivo validation of an MR-compatible temperature controllable superficial hyperthermia applicator for small animal studies

In this work, a superficial hyperthermia applicator was designed to be used in small animal studies. The applicator operates at 2.45 GHz and is compatible with a magnetic resonance scanner. A temperature controller was interfaced with the applicator to control the rate of change in temperature and the target temperature. Operation and robustness were validated through phantom and in vivo experiments

Quantification of DCE-MRI:A validation of three techniques with 3D-histology

Three different DCE-MRI quantification methods: model-free-based, compartment-model-based and principal component analysis, are compared by evaluating parameter maps for histological defined volumes of vital and non-vital tumor tissue. To obtain an accurate spatial correspondence between histology and DCE-MRI, a two-step registration process was used involving dense histological sampling, a reference plane and an intermediate ex vivo MRI. Results show that the model-free parameter washout and the second principal component score can adequately separate vital from non-vital tumor tissue, with an accuracy of respectively 99.2% and 99.7%. The other model-free parameters and the compartment-model-based Ktrans show some overlap in values between vital and non-vital tissue. The first, third and fourth pc-score have limited discriminative power.</p

Developing a tool for the validation of quantitative DCE-MRI

Dynamic Contrast-Enhanced Magnetic Resonance Imaging (DCE-MRI) is becoming an indispensable tool to non-invasively study tumor characteristics. However, many different DCE-analysis methods are currently being used. To compare and validate different methods, histology is the gold standard. For this purpose, exact co-localization between histology and MRI images is a prerequisite. In this study a methodology is developed to validate DCE-data with histology with an emphasis on correct registration of DCE-MRI and histological data. A pancreatic tumor was grown in a rat model. The tumor was dissected after MR imaging, embedded in paraffin, and cut into thin slices. These slices were stained with haematoxylin and eosin, digitized and stacked in a 3D volume. Next, the 3D histology was registered to exvivo SWI-weighted MR images, which in turn were registered to in-vivo SWI and DCE images to achieve correct colocalization. Semi-quantitative and quantitative parameters were calculated. Preliminary results suggest that both pharmacokinetic and heuristic DCE-parameters can discriminate between vital and non-vital tumor regions. The developed method offers the basis for an accurate spatial correlation between DCE-MRI derived parametric maps and histology, and facilitates the evaluation of different DCE-MRI analysis methods.</p

Inter and intra-tumor somatostatin receptor 2 heterogeneity influences peptide receptor radionuclide therapy response

Patients with neuroendocrine tumors (NETs) can be treated with peptide receptor radionuclide therapy (PRRT). Here, the somatostatin analogue octreotate radiolabeled with lutetium-177 is targeted to NET cells by binding to the somatostatin receptor subtype 2 (SST2). During radioactive decay, DNA damage is induced, leading to NET cell death. Although the therapy proves to be effective, mortality rates remain high. To appropriately select more optimal treatment strategies, it is essential to first better understand the radiobiological responses of tumor cells to PRRT. Methods: We analyzed PRRT induced radiobiological responses in SST2 expressing cells and xenografted mice using SPECT/MRI scanning and histological and molecular analyses. We measured [177Lu]Lu-DOTA-TATE uptake and performed analyses to visualize induction of DNA damage, cell death and other cellular characteristics. Results: The highest accumulation of radioactivity was measured in the tumor and kidneys. PRRT induced DNA damage signaling and repair in a time-dependent manner. We observed intra-tumor heterogeneity of DNA damage and apoptosis, which was not attributed to proliferation or bioavailability. We found a strong correlation between high DNA damage levels and high SST2 expression. PRRT elicited a different therapeutic response between models with different SST2 expression levels. Heterogeneous SST2 expression levels were also confirmed in patient NETs. Conclusion: Heterogeneous SST2 expression levels within NETs cause differentially induced DNA damage levels, influence recurrent tumor phenotypes and impact the therapeutic response in different models and potentially in patients. Our results contribute to a better understanding of PRRT effects, which might impact future therapeutic outcome of NET patients

Developing a tool for the validation of quantitative DCE-MRI

Dynamic Contrast-Enhanced Magnetic Resonance Imaging (DCE-MRI) is becoming an indispensable tool to non-invasively study tumor characteristics. However, many different DCE-analysis methods are currently being used. To compare and validate different methods, histology is the gold standard. For this purpose, exact co-localization between histology and MRI images is a prerequisite. In this study a methodology is developed to validate DCE-data with histology with an emphasis on correct registration of DCE-MRI and histological data. A pancreatic tumor was grown in a rat model. The tumor was dissected after MR imaging, embedded in paraffin, and cut into thin slices. These slices were stained with haematoxylin and eosin, digitized and stacked in a 3D volume. Next, the 3D histology was registered to exvivo SWI-weighted MR images, which in turn were registered to in-vivo SWI and DCE images to achieve correct colocalization. Semi-quantitative and quantitative parameters were calculated. Preliminary results suggest that both pharmacokinetic and heuristic DCE-parameters can discriminate between vital and non-vital tumor regions. The developed method offers the basis for an accurate spatial correlation between DCE-MRI derived parametric maps and histology, and facilitates the evaluation of different DCE-MRI analysis methods.</p