The purpose of this study was to investigate if radiomic analysis based on spectral micro-CT with nanoparticle contrastenhancement can differentiate tumors based on tumor-infiltrating lymphocyte (TIL) burden. High mutational load transplant soft tissue sarcomas were initiated in Rag2+/- and Rag2-/- mice to model varying TIL burden. Mice received radiation therapy (20 Gy) to the tumor-bearing hind limb and were injected with a liposomal iodinated contrast agent. Five days later, animals underwent conventional micro-CT imaging using an energy integrating detector (EID) and spectral micro-CT imaging using a photon-counting detector (PCD). Tumor volumes, and iodine uptakes were measured. The radiomic features (RF) were grouped into feature-spaces corresponding to EID, PCD, and spectral decomposition images. RFs were ranked to reduce redundancy and increase relevance based on TIL burden. A leave one out strategy was used to assess separation using a neural network classifier. Tumor iodine concentration was the only significantly different conventional tumor metric between Rag2+/- (TILs present) and Rag2-/- (TIL-deficient) tumors. RFs further enabled differentiation between Rag2+/- and Rag2-/- tumors. The PCD-derived RFs provided the highest accuracy (0.84) followed by decomposition-derived RFs (0.78) and the EID-derived RFs (0.65). Such non-invasive approaches could aid in tumor stratification for cancer therapy studies.
While the benefits of photon counting (PC) CT are significant, the performance of current PCDs is limited by physical effects distorting the spectral response. In this work, we examine a deep learning (DL) approach for spectral correction and material decomposition of PCCT data using multi-energy CT data sets acquired with an energy integrating detector (EID) for spectral calibration. We use a convolutional neural network (CNN) with a U-net structure and compare image domain and projection domain approaches to spectral correction and material decomposition. We trained our networks using noisy, spectrally distorted PCD projections as input data while the labels were derived from multi-energy EID data. For this study, we have scanned: 1) phantoms containing vials of iodine and calcium in water and 2) mice injected with an iodine-based liposomal contrast agent. Our results show that the image-based approach corrects best for spectral distortions and provides the lowest errors in material maps (RMSE in iodine: 0.34 mg/mL) compared with the projection-based approach (0.74 mg/mL) and image domain decomposition without correction (2.67 mg/mL). Both DL methods are, however, affected by a loss of spatial resolution (from 3 lp/mm in EID labels to ~2.2 lp/mm in corrected reconstructions). In summary, we demonstrate that multi-energy EID acquisitions can provide training data for DL-based spectral distortion correction. This data-driven correction does not require intimate knowledge of the spectral response or spectral distortions of the PCD. While the benefits of photon counting (PC) CT are significant, the performance of current PCDs is limited by physical effects distorting the spectral response. In this work, we examine a deep learning (DL) approach for spectral correction and material decomposition of PCCT data using multi-energy CT data sets acquired with an energy integrating detector (EID) for spectral calibration. We use a convolutional neural network (CNN) with a U-net structure and compare image domain and projection domain approaches to spectral correction and material decomposition. We trained our networks using noisy, spectrally distorted PCD projections as input data while the labels were derived from multi-energy EID data. For this study, we have scanned: 1) phantoms containing vials of iodine and calcium in water and 2) mice injected with an iodine-based liposomal contrast agent. Our results show that the image-based approach corrects best for spectral distortions and provides the lowest errors in material maps (RMSE in iodine: 0.34 mg/mL) compared with the projection-based approach (0.74 mg/mL) and image domain decomposition without correction (2.67 mg/mL). Both DL methods are, however, affected by a loss of spatial resolution (from 3 lp/mm in EID labels to ~2.2 lp/mm in corrected reconstructions). In summary, we demonstrate that multi-energy EID acquisitions can provide training data for DL-based spectral distortion correction. This data-driven correction does not require intimate knowledge of the spectral response or spectral distortions of the PCD.
We are developing imaging methods for the preclinical arm of a co-clinical trial investigating synergy between immunotherapy and radiotherapy. We perform in vivo micro-CT of mouse lungs to detect lung metastasis after treatment. This work explores deep learning (DL) as a fast and accurate approach to lung nodule segmentation. We examine the performance of DL lung tumor detection using realistically simulated nodules inserted into temporally-resolved real micro-CT datasets. Our simulations suggest that DL is a promising approach for fast, accurate segmentation of lung nodules in mice.
There is potential to improve CT imaging by adding spectral capabilities as given by photon counting detectors (PCD). Here we describe and assess performance of a new spectral micro-CT prototype system using a CdTe-based PCD with 100-μm pixel size (model XC-Thor, made by Direct Conversion) benefitting from anti-coincidence corrections. To assess the PCD in terms of spectral response, a Ba-133 nuclear source was scanned using full spectrum scanning by sweeping thresholds with 2 keV increments. In a different experiment, we used small vials containing water, iodine (I), gadolinium (Gd), and gold (Au) placed on the surface of the detector and acquired X-ray data in full spectrum mode to verify that the PCD threshold positions yielded the expected changes in contrast around the K edges of these elements. Detector performance was assessed using micro-CT phantoms and during cancer experiments in mice injected with nanoparticle (NP)-based contrast agents. Both phantom and mouse micro-CT data were reconstructed using our iterative, multi-channel algorithm based on the split Bregman method and regularization with rank-sparse kernel regression. A post-reconstruction decomposition method was used. The system is capable of high resolution (11.9 lp/mm, 10% MTF) tomographic imaging. Despite the anti-coincidence corrections, the spectral performance of the PCD is, however, not perfect, and it seems to be affected mostly at lower keVs, making accurate iodine decomposition challenging. Our cancer imaging results illustrate that our spectral micro-CT can benefit both nanotechnology and cancer research by providing an imaging method that can help test/optimize various nanoparticle for theranostics.
The maturation of photon-counting detector (PCD) technology promises to enhance routine CT applications with high-fidelity spectral information, transforming X-ray CT into a functional and molecular imaging modality, while maintaining the high spatial resolution, fast scanning times, and relatively low cost. We present our novel dual source pre-clinical micro-CT prototype system that combines a photon counting detector (PCD) and an energy integrating detector (EID) in a single system. We compare performance for PCD-only with combined hybrid (i.e. PCD and EID) CT imaging with equal dose. All CT data sets (EID-only, PCD-only, and hybrid) were reconstructed using our iterative, multi-channel algorithm based on the split Bregman method and regularization with rank-sparse kernel regression. We used a post-reconstruction decomposition method with iodine (I), gold (Au), gadolinium (Gd), and calcium (Ca) basis functions. Performance was assessed using micro-CT phantoms. Our results show the spatial resolution of the PCD-only and the hybrid reconstructions were similar and slightly better than for EID-only reconstruction with ~3.5 lp/mm (10% MTF). The NPS of the hybrid reconstruction was similar to PCD-only reconstruction. By co-reconstructing EID and PCD data, we achieved better image quality in the material decomposition but marginal differences in terms of concentration accuracy. Hybrid spectral micro- CT can benefit both nanotechnology and cancer research by providing imaging that can help test and optimize various NPs for theranostics.
Radiomics provide an exciting approach to developing imaging biomarkers in the context of precision medicine. We focus on the preclinical arm of a co-clinical trial investigating synergy of immunotherapy combined with radiation therapy (RT) and surgical resection using a genetically engineered mouse model of sarcoma. Our protocol involves the acquisition of MRI data with T1, T2 and T1 with contrast agent. There are two MRI time points i.e. one day before RT (20Gy) and one week later. After the second MRI acquisition the primary tumor is surgically removed, and the mice are followed for up to 6 months to investigate for local recurrence or distant metastases. The tumor images are segmented using deep learning. We performed radiomics for the tumor, peritumoral rim and the combined tumor and peritumoral rim. Our first radiomics analysis was focused on determining features which are most indicative to the effects of RT. Our second analysis aimed to answer if radiomics features could predict primary tumor recurrence within this population. Top features were selected for training classifiers based on neural networks and support vector machines. Our results show that gray level radiomic features show that tumors often acquire more heterogeneous texture and that tumor volume increases one-week post RT. The results also suggest that radiomics features serve to indicate likelihood of primary tumor recurrence with the best predictive power in the combined tumor and peritumoral area in pre-RT data (AUC: 0.83). In conclusion, we have created a radiomics pipeline to serve in our current preclinical arm of the co-clinical trial.
High-Z based nanoparticles (NP) are emerging as promising agents for both cancer radiotherapy (RT) and CT imaging. NPs can be delivered to tumors via the enhanced permeability and retention (EPR) effect and they preferentially accumulate in tumor’s perivascular region. Both gold and iodine NPs produce low-energy, short-range photoelectrons during RT, boosting radiation dose. Using spectral CT imaging, we sought to investigate (1) if iodine nanoparticles augmentation of RT increases vascular permeability in solid tumors, and (2) if iodine-RT induced changes in tumor vascular permeability improves delivery of nanoparticle-based chemotherapeutics. In vivo studies were performed in a carcinogen-induced and genetically engineered primary mouse model of soft tissue sarcoma. Tumor-bearing mice in test group were intravenously injected with liposomal-iodine (Lip-I) (1.32 g I/kg) on day 0. On day 1, both test (with Lip-I) and control (without Lip-I) mice received RT (single dose, 10 Gy). One day post-RT (day 2), all mice were intravenously injected with liposomal gadolinium (Lip-Gd) (0.32 g Gd/kg), a surrogate of nanoparticle chemotherapeutic agent. Three days later (day 5) mice were imaged on our hybrid spectral micro-CT system. A dual source pre-clinical CT prototype system that combines a photon counting detector (PCD) and an energy integrating detector (EID) in a single hybrid system served as our imaging device. The results demonstrate that Lip-I augmented RT, resulting in increased tumor vascular permeability compared to control mice treated with RT alone. Consequently, Lip-I +RT treated mice demonstrated a 4- fold higher intra-tumoral accumulation of Lip-Gd compared to RT alone treated mice. In conclusion, our work suggests that Lip-I augments RT-induced effects on tumor vasculature, resulting in increased vascular permeability and higher intratumoral deposition of chemotherapeutic nanoparticles.
Small animal imaging has become essential in evaluating new cancer therapies as they are translated from the preclinical to clinical domain. However, preclinical imaging is faced with unique challenges that emphasize the gap between mouse and man. One example is the difference in breathing patterns and breath-holding ability, which can dramatically affect tumor burden assessment in lung tissue. Our group is developing quantitative imaging methods for the preclinical arm of a co-clinical trial studying synergy between immunotherapy (anti-PD-1) and radiotherapy in a soft tissue sarcoma model. To mimic imaging performed in patients, primary sarcomas lesions are imaged with micro-MRI, while detection of lung metastases is performed with micro-CT. This study addresses whether respiratory gating during micro-CT acquisition improves lung tumor volume quantitation. Accuracy and precision of lung tumor measurements was determined by performing experiments involving simulations, a pocket phantom and in vivo scans with and without prospective respiratory gating. Sensitivity and precision of segmentation with and without gating was studied using simulated lung tumors. A clinically-inspired “pocket phantom” was used during in vivo mouse scanning to aid in refining and assessing the gating protocols. Finally, we performed a series of in vivo scans on tumor-bearing mice while varying the animal’s position (test-retest), and performing the analyses in triplicate to assess the effects of gating. Application of respiratory gating techniques reduced variance of repeated volume measurements and significantly improved the accuracy of tumor volume quantitation in vivo.
Small animal imaging is essential in building a bridge from basic science to the clinic by providing the confidence necessary to move new cancer therapies to patients. However, there is considerable variability in preclinical imaging, including tumor volume estimations based on tumor segmentation procedures which can be clearly user-biased. Our group is engaged in developing quantitative imaging methods which will be applied in the preclinical arm of a co-clinical trial studying synergy between anti-PD-1 treatment and radiotherapy using a genetically engineered mouse model of soft tissue sarcoma. This study focuses on a convolutional neural network (CNN)-based method for automatic tumor segmentation based on multimodal MRI images, i.e. T1 weighted, T2 weighted and T1 weighted with contrast agent. Our images were acquired on a 7.0 T Bruker Biospec small animal MRI scanner. Preliminary results show that our U-net structure and 3D patch-wise approach using both Dice and cross entropy loss functions delivers strong segmentation results. We have also compared single performance using only T2 weighted versus multimodal MR images for CNN segmentation. Our results showthat Dice similarity coefficient were higher when using multimodal versus single T2 weighted data (0.84 ± 0.05 and 0.81 ± 0.03). In conclusion, we successfully established a segmentation method for preclinical MR sarcoma data based on deep learning. This approach has the advantage of reducing user bias in tumor segmentation and improving the accuracy and precision of tumor volume estimations for co-clinical cancer trials.
Spectral computed tomography (CT) using photon counting detectors (PCDs) can provide accurate tissue composition measurements by utilizing the energy dependence of x-ray attenuation in different materials. PCDs are especially suited for K-edge imaging, revealing the spatial distribution of select imaging probes through quantitative material decomposition. We report on a prototype spectral micro-CT system with a CZT-based PCD (DxRay, Inc.) that has 16 × 16 pixels of 0.5 × 0.5 mm2, a thickness of 3 mm, and four energy thresholds. Due to the PCD’s limited size (8 × 8 mm2), our system uses a translate-rotate projection acquisition strategy to cover a field of view relevant for preclinical imaging (∼4.5 cm). Projection corrections were implemented to minimize artifacts associated with dead pixels and projection stitching. A sophisticated iterative algorithm was used to reconstruct both phantom and ex vivo mouse data. To achieve preclinically relevant spatial resolution, we trained a convolutional neural network to perform pan-sharpening between low-resolution PCD data (247-μm voxels) and high-resolution energy-integrating detector data (82-μm voxels), recovering a high-resolution estimate of the spectral contrast suitable for material decomposition. Long-term, preclinical spectral CT systems such as ours could serve in the developing field of theranostics (therapy and diagnostics) for cancer research.
Advances in CT hardware have propelled the development of novel CT contrast agents. Combined with the spectral capabilities of X-ray CT, molecular imaging is possible using multiple heavy-metal contrast agents. Nanoparticle platforms make particularly attractive agents because of (1) their ability to carry a large payload of imaging moieties, and (2) their ease of surface modification to facilitate molecular targeting. While several novel imaging moieties based on high atomic number elements are being explored, iodine (I) and gadolinium (Gd) are particularly attractive because they are already in clinical use. In this work, we investigate the feasibility for in vivo discrimination of iodine and gadolinium nanoparticles using dual energy micro-CT. Phantom experiments were performed to measure the CT enhancement for I and Gd over a range of voltages from 40 to 80 kVp using a dual-source micro-CT system with energy integrating detectors having cesium iodide scintillators. The two voltages that provide maximum discrimination between I and Gd were determined to be 50 kVp with Cu filtration and 40 kVp without any filtration. Serial dilutions of I and Gd agents were imaged to determine detection sensitivity using the optimal acquisition parameters. Next, an in vivo longitudinal small animal study was performed using Liposomal I (Lip-I) and Liposomal Gd (Lip-Gd) nanoparticles. The mouse was intravenously administered Lip-Gd and imaged within 1 h post-contrast to visualize Gd in the vascular compartment. The animal was reimaged at 72 h post-contrast with dual-energy micro-CT at 40 kVp and 50 kVp to visualize the accumulation of Lip-Gd in the liver and spleen. Immediately thereafter, the animal was intravenously administered Lip-I and re-imaged. The dual energy sets were used to estimate the concentrations of Gd and I via a two-material decomposition with a non-negativity constraint. The phantom results indicated that the relative contrast enhancement per mg/ml of I to Gd was 0.85 at 40 kVp and 1.79 at 50 kVp. According to the Rose criterion (CNR<5), the detectability limits were 2.67 mg/ml for I and 2.46 mg/ml for Gd. The concentration maps confirmed the expected biodistribution, with Gd concentrated in the spleen and with I in the vasculature of the kidney, liver, and spleen. Iterative reconstruction provided higher sensitivity to detect relatively low concentrations of gadolinium. In conclusion, dual energy micro-CT can be used to discriminate and simultaneously image probes containing I and Gd.
Spectral CT can provide accurate tissue composition measurements by utilizing the energy dependence of x-ray attenuation in different materials. We have introduced image reconstruction and material decomposition algorithms for multi-energy CT data acquired either with energy integrating detectors (EID) or photon counting detectors (PCD); however, material decomposition is an ill-posed problem due to the potential overlap of spectral measurements and to noise. Recently, convolutional neural networks (CNN) have generated excitement in the field of machine learning and computer vision. The goal of this work is to develop CNN-based methods for material decomposition in spectral CT. The CNN for decomposition had a U-net structure and was trained with either five-energy PCD-CT or DE-CT. As targets for training, we used simulated phantoms constructed from random combinations of water and contrast agents (iodine, barium, and calcium for five-energy PCD-CT; iodine and gold for DE EID-based CT). The experimentally measured sensitivity matrix values for iodine, barium, and calcium or iodine and gold were used to recreate the CT images corresponding to both PCD and DE-CT cases. These CT images were used to train CNNs to generate material maps at each pixel location. After training, we tested the CNNs by applying them to experimentally acquired DE-EID and PCD-based micro-CT data in mice. The predicted material maps were compared to the absolute truth in simulations and to sensitivity-based decompositions for the in vivo mouse data. The CNN-based decomposition provided higher accuracy and lower noise. In conclusion, our U-net performed a more robust spectral micro-CT decomposition because it inherently better exploits spatial and spectral correlations.
Dual energy (DE) micro-CT shows great potential to provide accurate tissue composition by utilizing the energy dependence of x-ray attenuation in different materials. This is especially well-suited for pre-clinical imaging using nanoparticle-based contrast agents in situations where quantitative material decomposition helps probe processes which are otherwise limited by poor soft tissue contrast. We have previously proposed optimal in vivo DE micro-CT methods for imaging using iodinated and gold nanoparticles. However, in vivo studies are limited in spatial resolution due to constraints in sampling time and radiation dose. Ex vivo dual energy imaging can provide much higher resolution and can serve as a validation of in vivo studies. Our study proposes multiscale in vivo and ex vivo DE micro-CT of the same subjects using two in-house developed micro-CT systems. We use a dual source micro-CT system to scan a mouse that has been injected with both iodinated and gold nanoparticles for in vivo DE scanning at 63 micron resolution. The same mouse is then scanned ex vivo with DE on a separate single source micro-CT system at a spatial resolution of 22 microns. We perform reconstructions using filtered back projection followed by noise reduction via joint bilateral filtration. A dynamic flat field correction method has been applied on the ex vivo micro-CT data to correct for image artifacts. A DE post-reconstruction decomposition is used to create iodine and gold material maps which are used to measure accumulation of contrast agent within the body. We evaluate challenges associated with each imaging methodology. Our results compare image quality and material maps. Overall, our methods represent a substantial tool for multiscale DE micro-CT imaging using wellcharacterized contrast agents and serving various applications in biological research.
Spectral CT using photon counting x-ray detectors (PCXDs) can provide accurate tissue composition measurements by utilizing the energy dependence of x-ray attenuation in different materials. PCXDs are especially suited for imaging Kedge contrast agents, revealing the spatial distribution of select imaging probes through quantitative material decomposition. To further advance the field, there is a clear and continuing need to develop PCXD hardware and software as part of a new generation of spectral CT imaging systems. Our group specializes in the development of preclinical microCT systems and of novel imaging probes based on K-edge materials. Toward this goal, we have now developed a prototype spectral micro-CT system with a PCXD produced by DxRay. This CZT-based PCXD has 16x16 pixels, each with a size of 0.5 x 0.5 mm, a thickness of 3 mm, and 4 configurable energy thresholds. The detector is thus only 8 mm x 8 mm in size. Due to the limited size of this detector tile, we have implemented a translate-rotate micro-CT system (i.e. a 2nd generation scanner). In this paper we summarize considerable efforts which went into compensating for dead pixels and for pixels with non-linear responses to prevent artifacts in the CT reconstruction results. We also present spectral response measurements for the detector and the results of both phantom and animal experiments with iodine- and gold-based contrast agents. The results confirm our ability to sample and reconstruct tomographic images, but also show that the PCXD prototype has limitations in imaging iodine.
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