Deletion of the mRNA stability factor ELAVL1 (HuR) in pancreatic cancer cells disrupts the tumor microenvironment integrity.

Stromal cells promote extensive fibrosis in pancreatic ductal adenocarcinoma (PDAC), which is associated with poor prognosis and therapeutic resistance. We report here for the first time that loss of the RNA-binding protein human antigen R (HuR, ELAVL1) in PDAC cells leads to reprogramming of the tumor microenvironment. In multiple in vivo models, CRISPR deletion of ELAVL1 in PDAC cells resulted in a decrease of collagen deposition, accompanied by a decrease of stromal markers (i.e. podoplanin, α-smooth muscle actin, desmin). RNA-sequencing data showed that HuR plays a role in cell-cell communication. Accordingly, cytokine arrays identified that HuR regulates the secretion of signaling molecules involved in stromal activation and extracellular matrix organization [i.e. platelet-derived growth factor AA (PDGFAA) and pentraxin 3]. Ribonucleoprotein immunoprecipitation analysis and transcription inhibition studies validated PDGFA mRNA as a novel HuR target. These data suggest that tumor-intrinsic HuR supports extrinsic activation of the stroma to produce collagen and desmoplasia through regulating signaling molecules (e.g. PDGFAA). HuR-deficient PDAC in vivo tumors with an altered tumor microenvironment are more sensitive to the standard of care gemcitabine, as compared to HuR-proficient tumors. Taken together, we identified a novel role of tumor-intrinsic HuR in its ability to modify the surrounding tumor microenvironment and regulate PDGFAA.

An Advanced Thermal Decomposition Method to Produce Magnetic Nanoparticles with Ultrahigh Heating Efficiency for Systemic Magnetic Hyperthermia.

Due to the limited heating efficiency of available magnetic nanoparticles, it is difficult to achieve therapeutic temperatures above 44 °C in relatively inaccessible tumors during magnetic hyperthermia following systemic administration of nanoparticles at clinical dosage (≤10 mg kg-1 ). To address this, a method for the preparation of magnetic nanoparticles with ultrahigh heating capacity in the presence of an alternating magnetic field (AMF) is presented. The low nitrogen flow rate of 10 mL min-1 during the thermal decomposition reaction results in cobalt-doped nanoparticles with a magnetite (Fe3 O4 ) core and a maghemite (γ-Fe2 O3 ) shell that exhibit the highest intrinsic loss power reported to date of 47.5 nH m2 kg-1 . The heating efficiency of these nanoparticles correlates positively with increasing shell thickness, which can be controlled by the flow rate of nitrogen. Intravenous injection of nanoparticles at a low dose of 4 mg kg-1 elevates intratumoral temperatures to 50 °C in mice-bearing subcutaneous and metastatic cancer grafts during exposure to AMF. This approach can also be applied to the synthesis of other metal-doped nanoparticles with core-shell structures. Consequently, this method can potentially be used for the development of novel nanoparticles with high heating performance, further advancing systemic magnetic hyperthermia for cancer treatment.

Pharmacological reduction of coagulation factor XI reduces macrophage accumulation and accelerates deep vein thrombosis resolution in a mouse model of venous thrombosis.

BACKGROUND: Deep vein thrombosis (DVT) and post-thrombotic syndrome (PTS) remain highly prevalent despite modern medical therapy. Contact activation is a promising target for safe antithrombotic anticoagulation. The anti-factor XI (FXI) monoclonal antibody 14E11 reduces circulating levels of FXI without compromising hemostasis. The human recombinant analog, AB023, is in clinical development. The role of FXI in mediation of inflammation during DVT resolution is unknown. OBJECTIVES: Investigate the effects of pharmacological targeting of FXI with 14E11 in an experimental model of venous thrombosis. METHODS: Adult wild-type CD1 mice were treated with subcutaneous anti-FXI antibody (14E11, 5 mg/kg) versus saline prior to undergoing surgical constriction of the inferior vena cava (IVC). Mice were evaluated at various time points to assess thrombus weight and volume, as well as histology analysis, ferumoxytol enhanced magnetic resonance imaging (Fe-MRI), and whole blood flow cytometry. RESULTS: 14E11-treated mice had reduced thrombus weights and volumes after IVC constriction on day 7 compared to saline-treated mice. 14E11 treatment reduced circulating monocytes by flow cytometry and macrophage content within thrombi as evaluated by histologic staining and Fe-MRI. Collagen deposition was increased at day 3 while CD31 and smooth muscle cell actin expression was increased at day 7 in the thrombi of 14E11-treated mice compared to saline-treated mice. CONCLUSION: Pharmacologic targeting of FXI enhances the early stages of experimental venous thrombus resolution in wild-type CD1 mice, and may be of interest for future clinical evaluation of the antibody in DVT and PTS.

Targeted Nanoparticles with High Heating Efficiency for the Treatment of Endometriosis with Systemically Delivered Magnetic Hyperthermia.

Endometriosis is a devastating disease in which endometrial-like tissue forms lesions outside the uterus. It causes infertility and severe pelvic pain in ≈176 million women worldwide, and there is currently no cure for this disease. Magnetic hyperthermia could potentially eliminate widespread endometriotic lesions but has not previously been considered for treatment because conventional magnetic nanoparticles have relatively low heating efficiency and can only provide ablation temperatures (>46 °C) following direct intralesional injection. This study is the first to describe nanoparticles that enable systemically delivered magnetic hyperthermia for endometriosis treatment. When subjected to an alternating magnetic field (AMF), these hexagonal iron-oxide nanoparticles exhibit extraordinary heating efficiency that is 6.4× greater than their spherical counterparts. Modifying nanoparticles with a peptide targeted to vascular endothelial growth factor receptor 2 (VEGFR-2) enhances their endometriosis specificity. Studies in mice bearing transplants of macaque endometriotic tissue reveal that, following intravenous injection at a low dose (3 mg per kg), these nanoparticles efficiently accumulate in endometriotic lesions, selectively elevate intralesional temperature above 50 °C upon exposure to external AMF, and completely eradicate them with a single treatment. These nanoparticles also demonstrate promising potential as magnetic resonance imaging (MRI) contrast agents for precise detection of endometriotic tissue before AMF application.

3D MR fingerprinting using Seiffert spirals.

PURPOSE: Seiffert spirals were recently explored as an efficient way to traverse 3D k-space compared to traditional 3D techniques. Several studies have shown the ability of 3D MR fingerprinting (MRF) techniques to acquire T1 and T2 relaxation maps in a short period of time. However, these sequences do not sample across a large region of 3D k-space every TR, especially in the way that Seiffert trajectories can. METHODS: A 3D MRF sequence was designed using 8 Seiffert spirals rotated in 3D k-space, with flip angle modulation for T1 and T2 sensitivity. The sequence was compared to an MRF sequence using a 2D spiral rotated in 3D k-space using the tiny golden angle acquisition with similar resolution/readout duration. Both sequences were evaluated using simulations, phantom validation, and in vivo imaging. RESULTS: In all experiments, the Seiffert spiral MRF sequence performed similar to if not better than the multi-axis 2D spiral MRF sequence. Strong intraclass correlation coefficients (> 0.9) were found between conventional and MRF sequences in phantoms, whereas the in vivo results showed slightly less aliasing artifact with the Seiffert trajectory. CONCLUSION: In this study, Seiffert spirals were used within the MRF framework to acquire high-resolution T1 and T2 relaxation time maps in less than 2.5 min. The reduced aliasing artifacts seen with the Seiffert sequence suggests that sampling over 3D k-space evenly each TR can improve quantification or shorten scan times.

T1ρ magnetic resonance fingerprinting.

T1ρ relaxation imaging is a quantitative imaging technique that has been used to assess cartilage integrity, liver fibrosis, tumors, cardiac infarction, and Alzheimer's disease. T1 , T2 , and T1ρ relaxation time constants have each demonstrated different degrees of sensitivity to several markers of fibrosis and inflammation, allowing for a potential multi-parametric approach to tissue quantification. Traditional magnetic resonance fingerprinting (MRF) has been shown to provide quick, quantitative mapping of T1 and T2 relaxation time constants. In this study, T1ρ relaxation is added to the MRF framework using spin lock preparations. An MRF sequence involving an RF-spoiled sequence with TR , flip angle, T1ρ , and T2 preparation variation is described. The sequence is then calibrated against conventional T1 , T2 , and T1ρ relaxation mapping techniques in agar phantoms and the abdomens of four healthy volunteers. Strong intraclass correlation coefficients (ICC > 0.9) were found between conventional and MRF sequences in phantoms and also in healthy volunteers (ICC > 0.8). The highest ICC correlation values were seen in T1 , followed by T1ρ and then T2 . In this study, T1ρ relaxation has been incorporated into the MRF framework by using spin lock preparations, while still fitting for T1 and T2 relaxation time constants. The acquisition of these parameters within a single breath hold in the abdomen alleviates the issues of movement between breath holds in conventional techniques.

Multi-parametric T2 * magnetic resonance fingerprinting using variable echo times.

The use of quantitative imaging biomarkers in the imaging of various disease states, including cancer and neurodegenerative disease, has increased in recent years. T1 , T2 , and T2 * relaxation time constants have been shown to be affected by tissue structure or contrast infusion. Acquiring these biomarkers simultaneously in a multi-parametric acquisition could provide more robust detection of tissue changes in various disease states including neurodegeneration and cancer. Traditional magnetic resonance fingerprinting (MRF) has been shown to provide quick, quantitative mapping of T1 and T2 relaxation time constants. In this study, T2 * relaxation is added to the MRF framework using variable echo times (TE). To demonstrate the feasibility of the method and compare incremental and golden angle spiral rotations, simulated phantom data was fit using the proposed method. Additionally, T1 /T2 /T2 */δf MRF as well as conventional T1 , T2 , and T2 * acquisitions were acquired in agar phantoms and the brains of three healthy volunteers. Golden angle spiral rotation was found to reduce inaccuracy resulting from off resonance effects. Strong correlations were found between conventional and MRF values in the T1 , T2 , and T2 * relaxation time constants of the agar phantoms and healthy volunteers. In this study, T2 * relaxation has been incorporated into the MRF framework by using variable echo times, while still fitting for T1 and T2 relaxation time constants. In addition to fitting these relaxation time constants, a novel method for fitting and correcting off resonance effects has been developed.

Scoring hip osteoarthritis with MRI (SHOMRI): A whole joint osteoarthritis evaluation system.

BACKGROUND: To develop a semi-quantitative MR-based hip osteoarthritis (OA) evaluation system (Scoring hip osteoarthritis with MRI, SHOMRI), and to test its reproducibility and face validity. METHODS: The study involved 98 subjects with informed consent. Three-Tesla MR imaging of hip was performed in three planes with intermediate-weighted fat saturated FSE sequences. Two radiologists assessed cartilage loss, bone marrow edema pattern, subchondral cyst in 10 subregions, and assessed labrum in 4 subregions. In addition, presence or absence of ligamentum teres integrity, paralabral cysts, intra-articular body, and effusion in the hip joint were analyzed using the SHOMRI system. The reproducibility was assessed with intra-class correlation coefficient (ICC), Cohen's Kappa values and percent agreement. SHOMRI scores were correlated with radiographic Kellgren-Lawrence (KL) and OARSI atlas gradings, and clinical parameters, the hip osteoarthritis outcome score (HOOS) and hip range of motion (ROM), using Spearman's rank correlation and ordinal logistic regression. RESULTS: ICC values were in the excellent range, 0.91 to 0.97. Cohen's Kappa values and percent agreement ranged from 0.55 to 0.79 and 66 to 99%, respectively. SHOMRI demonstrated significant correlations with KL and OARSI gradings as well as with clinical parameters, HOOS and ROM (P < 0.05). Among the SHOMRI features, subchondral cyst and bone marrow edema pattern showed the highest correlation with HOOS and ROM. CONCLUSION: SHOMRI demonstrated moderate to excellent reproducibility and significant correlation with radiographic gradings and clinical parameters.

Correction of breathing-induced errors in magnetic resonance thermometry of hyperthermia using multiecho field fitting techniques.

PURPOSE: Breathing motion can create large errors when performing magnetic resonance (MR) thermometry of the breast. Breath holds can be used to minimize these errors, but not eliminate them. Between breath holds, the referenceless method can be used to further reduce errors by relying on regions of nonheated fatty tissue surrounding the heated region. When the surrounding tissue is heated (i.e., for a hyperthermia treatment), errors can result due to phase changes of the small amounts of water in the tissue. Therefore, an extension of the referenceless method is proposed which fits for the field in fatty tissue independent of temperature change and extrapolates it to the water-rich regions. METHODS: Nonheating experiments were performed with male volunteers performing breath holds on top of a phantom mimicking a breast with a tumor. Heating experiments were also conducted with the same phantom while mechanically simulated breath holds were performed. A nonheating experiment was also performed with a healthy female breast. For each experiment, a nonlinear fitting algorithm was used to fit for temperature change and B0 field inside of the fatty tissue. The field changes were then extrapolated into water-rich (tumor) portions of the image using a least-squares fit to a fifth-order equation, to correct for field changes due to breath hold changes. Similar results were calculated using the image phase, to mimic the use of the referenceless method. RESULTS: Phantom results showed large reduction of mean error and standard deviation. In the non-heating experiments, the traditional referenceless method and our extended method both corrected by similar amounts. However, in the heating experiments, the average deviation of the temperature calculated with the extended method from a fiber optic probe temperature was approximately 50% less than the deviation with the referenceless method. The in vivo breast results demonstrated reduced standard deviation and mean. CONCLUSIONS: In this paper, we have developed an extension of the referenceless method to correct for breathing errors using multiecho fitting methods to fit for the B0 field in the fatty tissue and using measured field changes as references to extrapolate field corrections into a water-only (tumor) region. This technique has been validated in a number of situations, and in all cases, the correction method has been shown to greatly reduce temperature error in water-rich regions. The method has also been shown to be an improvement over similar methods that use image phase changes instead of field changes, particularly when temperature changes are induced.

Accuracy of real time noninvasive temperature measurements using magnetic resonance thermal imaging in patients treated for high grade extremity soft tissue sarcomas.

PURPOSE: To establish accuracy of real time noninvasive temperature measurements using magnetic resonance thermal imaging in patients treated for high grade extremity soft tissue sarcomas. METHODS: Protocol patients with advanced extremity sarcomas were treated with external beam radiation therapy and hyperthermia. Invasive temperature measures were compared to noninvasive magnetic resonance thermal imaging (MRTI) at 1.5 T performed during hyperthermia. Volumetric temperature rise images were obtained using the proton resonance frequency shift (PRFS) technique during heating in a 140 MHz miniannular phased array applicator. MRTI temperature changes were compared to invasive measurements of temperature with a multisensor fiber optic probe inside a #15 g catheter in the tumor. Since the PRFS technique is sensitive to drifts in the primary imaging magnetic field, temperature change distributions were corrected automatically during treatment using temperature-stable reference materials to characterize field changes in 3D. The authors analyzed MRT images and compared, in evaluable treatments, MR-derived temperatures to invasive temperatures measured in extremity sarcomas. Small regions of interest (ROIs) were specified near each invasive sensor identified on MR images. Temperature changes in the interstitial sensors were compared to the corresponding ROI PRFS-based temperature changes over the entire treatment and over the steady-state period. Nonevaluable treatments (motion/imaging artifacts, noncorrectable drifts) were not included in the analysis. RESULTS: The mean difference between MRTI and interstitial probe measurements was 0.91 degrees C for the entire heating time and 0.85 degrees C for the time at steady state. These values were obtained from both tumor and normal tissue ROIs. When the analysis is done on just the tumor ROIs, the mean difference for the whole power on time was 0.74 degrees C and during the period of steady state was 0.62 degrees C. CONCLUSIONS: The data show that for evaluable treatments, excellent correlation (deltaT < 1 degrees C) of MRTI-ROI and invasive measurements can be achieved, but that motion and other artifacts are still serious challenges that must be overcome in future work.