Hyperpolarized [1-13C]-pyruvate MRS evaluates immune potential and predicts response to radiotherapy in cervical cancer.

BACKGROUND: Monitoring pyruvate metabolism in the spleen is important for assessing immune activity and achieving successful radiotherapy for cervical cancer due to the significance of the abscopal effect. We aimed to explore the feasibility of utilizing hyperpolarized (HP) [1-13C]-pyruvate magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) to evaluate pyruvate metabolism in the human spleen, with the aim of identifying potential candidates for radiotherapy in cervical cancer. METHODS: This prospective study recruited six female patients with cervical cancer (median age 55 years; range 39-60) evaluated using HP [1-13C]-pyruvate MRI/MRS at baseline and 2 weeks after radiotherapy. Proton (1H) diffusion-weighted MRI was performed in parallel to estimate splenic cellularity. The primary outcome was defined as tumor response to radiotherapy. The Student t-test was used for comparing 13C data between the groups. RESULTS: The splenic HP [1-13C]-lactate-to-total carbon (tC) ratio was 5.6-fold lower in the responders than in the non-responders at baseline (p = 0.009). The splenic [1-13C]-lactate-to-tC ratio revealed a 1.7-fold increase (p = 0.415) and the splenic [1-13C]-alanine-to-tC ratio revealed a 1.8-fold increase after radiotherapy (p = 0.482). The blood leukocyte differential count revealed an increased proportion of neutrophils two weeks following treatment, indicating enhanced immune activity (p = 0.013). The splenic apparent diffusion coefficient values between the groups were not significantly different. CONCLUSIONS: This exploratory study revealed the feasibility of HP [1-13C]-pyruvate MRS of the spleen for evaluating baseline immune potential, which was associated with clinical outcomes of cervical cancer after radiotherapy. TRIAL REGISTRATION: ClinicalTrials.gov NCT04951921 , registered 7 July 2021. RELEVANCE STATEMENT: This prospective study revealed the feasibility of using HP 13C MRI/MRS for assessing pyruvate metabolism of the spleen to evaluate the patients' immune potential that is associated with radiotherapeutic clinical outcomes in cervical cancer. KEY POINTS: • Effective radiotherapy induces abscopal effect via altering immune metabolism. • Hyperpolarized 13C MRS evaluates patients' immune potential non-invasively. • Pyruvate-to-lactate conversion in the spleen is elevated following radiotherapy.

Comparing Magnetic Resonance Fingerprinting (MRF) and the MAGiC Sequence for Simultaneous T1 and T2 Quantitative Measurements in the Female Pelvis: A Prospective Study.

The aim of this study was to explore the potential of magnetic resonance fingerprinting (MRF), an emerging quantitative MRI technique, in measuring relaxation values of female pelvic tissues compared to the conventional magnetic resonance image compilation (MAGiC) sequence. The study included 32 female patients who underwent routine pelvic MRI exams using anterior and posterior array coils on a 3T clinical scanner. Our findings demonstrated significant correlations between MRF and MAGiC measured T1 and T2 values (p < 0.0001) for various pelvic tissues, including ilium, femoral head, gluteus, obturator, iliopsoas, erector spinae, uterus, cervix, and cutaneous fat. The tissue contrasts generated from conventional MRI and synthetic MRF also showed agreement in bone, muscle, and uterus for both T1-weighted and T2-weighted images. This study highlights the strengths of MRF in providing simultaneous T1 and T2 mapping. MRF offers distinct tissue contrast and has the potential for accurate diagnosis of female pelvic diseases, including tumors, fibroids, endometriosis, and pelvic inflammatory disease. Additionally, MRF shows promise in monitoring disease progression or treatment response. Overall, the study demonstrates the potential of MRF in the field of female pelvic organ imaging and suggests that it could be a valuable addition to the clinical practice of pelvic MRI exams. Further research is needed to establish the clinical utility of MRF and to develop standardized protocols for its implementation in clinical practice.

Developing a Method to Estimate the Downstream Metabolite Signals from Hyperpolarized [1-13C]Pyruvate.

Hyperpolarized carbon-13 MRI has the advantage of allowing the study of glycolytic flow in vivo or in vitro dynamically in real-time. The apparent exchange rate constant of a metabolite dynamic signal reflects the metabolite changes of a disease. Downstream metabolites can have a low signal-to-noise ratio (SNR), causing apparent exchange rate constant inconsistencies. Thus, we developed a method that estimates a more accurate metabolite signal. This method utilizes a kinetic model and background noise to estimate metabolite signals. Simulations and in vitro studies with photon-irradiated and control groups were used to evaluate the procedure. Simulated and in vitro exchange rate constants estimated using our method were compared with the raw signal values. In vitro data were also compared to the Area-Under-Curve (AUC) of the cell medium in 13C Nuclear Magnetic Resonance (NMR). In the simulations and in vitro experiments, our technique minimized metabolite signal fluctuations and maintained reliable apparent exchange rate constants. In addition, the apparent exchange rate constants of the metabolites showed differences between the irradiation and control groups after using our method. Comparing the in vitro results obtained using our method and NMR, both solutions showed consistency when uncertainty was considered, demonstrating that our method can accurately measure metabolite signals and show how glycolytic flow changes. The method enhanced the signals of the metabolites and clarified the metabolic phenotyping of tumor cells, which could benefit personalized health care and patient stratification in the future.

Magnetic Resonance Spectroscopy for Risk Stratification of Sonographically Indeterminate Ovarian Neoplasms: Preliminary Study.

We aim to assess the additional value of diffusion-weighted imaging (DWI) and magnetic resonance spectroscopy (MRS) for the risk stratification of sonographically indeterminate ovarian neoplasms. A total of 21 patients with diagnosed adnexal masses between 2014 and 2017 were divided into malignant (four serous cystadenocarcinomas, four endometrioid carcinomas, three clear cell carcinomas, and one carcinosarcoma) and benign (four cystadenomas, two teratomas, one fibroma, one endometrioma, and one corpus luteal cyst) groups. An apparent diffusion coefficient (ADC) value of 1.27 × 10-3 mm2/s was considered as the optimal threshold in distinguishing malignant from benign ovarian tumors (sensitivity and specificity: 100% and 77.8%, respectively). Choline peaks were detected in six of seven O-RADS (Ovarian-Adnexal Imaging-Reporting Data System) 4 lesions and corrected all of the DWI false-negative clear cell carcinoma. Based on the presence of the choline peaks, the diagnostic performance of MRS showed a sensitivity of 77.8%, a specificity of 100%, and an accuracy of 85.7%, respectively. In conclusion, MRS could potentially play a complementary role for DWI in tumor characterization, particularly for O-RADS 4 tumors or clear cell carcinomas.

Monitoring Early Glycolytic Flux Alterations Following Radiotherapy in Cancer and Immune Cells: Hyperpolarized Carbon-13 Magnetic Resonance Imaging Study.

Alterations in metabolism following radiotherapy affect therapeutic efficacy, although the mechanism underlying such alterations is unclear. A new imaging technique-named dynamic nuclear polarization (DNP) carbon-13 magnetic resonance imaging (MRI)-probes the glycolytic flux in a real-time, dynamic manner. The [1-13C]pyruvate is transported by the monocarboxylate transporter (MCT) into cells and converted into [1-13C]lactate by lactate dehydrogenase (LDH). To capture the early glycolytic alterations in the irradiated cancer and immune cells, we designed a preliminary DNP 13C-MRI study by using hyperpolarized [1-13C]pyruvate to study human FaDu squamous carcinoma cells, HMC3 microglial cells, and THP-1 monocytes before and after irradiation. The pyruvate-to-lactate conversion rate (kPL [Pyr.]) calculated by kinetic modeling was used to evaluate the metabolic alterations. Western blotting was performed to assess the expressions of LDHA, LDHB, MCT1, and MCT4 proteins. Following irradiation, the pyruvate-to-lactate conversion rates on DNP 13C-MRI were significantly decreased in the FaDu and the HMC3 cells but increased in the THP-1 cells. Western blot analysis confirmed the similar trends in LDHA and LDHB expression levels. In conclusion, DNP 13C-MRI non-invasively captured the different glycolytic alterations among cancer and immune systems in response to irradiation, implying its potential for clinical use in the future.

Exploring Potential Carcinogenic Activity of Per- and Polyfluorinated Alkyl Substances Utilizing High-Throughput Toxicity Screening Data.

Per- and polyfluorinated alkyl substances (PFAS) are ubiquitous, persistent, and toxic chemicals that pose public health risks. Recent carcinogenicity concerns have arisen based on epidemiological studies, animal tumor findings, and mechanistic data. Thousands of PFAS exist; however, current understanding of their toxicity is informed by studies of a select few, namely, perfluorooctanoic acid and perfluorooctanesulfonic acid. Hence, the computational, high-throughput screening tool, the US EPA CompTox Chemical Dashboard's ToxCast, was utilized to explore the carcinogenicity potential of PFAS. Twenty-three major PFAS that had sufficient in vitro ToxCast data and covered a range of structural subclasses were analyzed with the visual analytics software ToxPi, yielding a qualitative and quantitative assessment of PFAS activity in realms closely linked with carcinogenicity. A comprehensive literature search was also conducted to check the consistency of analyses with other mechanistic data streams. The PFAS were found to induce a vast range of biological perturbations, in line with several of the International Agency for Research on Cancer-defined key carcinogen characteristics. Patterns observed varied by length of fluorine-bonded chains and/or functional group within and between each key characteristic, suggesting some structure-based variability in activity. In general, the major conclusions drawn from the analysis, that is, the most notable activities being modulation of receptor-mediated effects and induction of oxidative stress, were supported by literature findings. The study helps enhance understanding of the mechanistic pathways that underlie the potential carcinogenicity of various PFAS and hence could assist in hazard identification and risk assessment for this emerging and relevant class of environmental toxicants.

Evaluation of chemotherapeutic and cancer-protective properties of sphingosine and C2-ceramide in a human breast stem cell derived carcinogenesis model.

The overall goal of the present study was to evaluate the chemotherapeutic and cancer­protective properties of D­erythro­sphingosine (sphingosine) and C2­ceramide using a human breast epithelial cell (HBEC) culture system, which represents multiple­stages of breast carcinogenesis. The HBEC model includes Type I HBECs (normal stem), Type II HBECs (normal differentiated) and transformed cells (immortal/non­tumorigenic cells and tumorigenic cells, which are transformed from the same parental normal stem cells). The results of the present study indicate that sphingosine preferentially inhibits proliferation and causes death of normal stem cells (Type I), tumorigenic cells, and MCF7 breast cancer cells, but not normal differentiated cells (Type II). In contrast to the selective anti­proliferative effects of sphingosine, C2­ceramide inhibits proliferation of normal differentiated cells as well as normal stem cells, tumorigenic cells, and MCF7 cancer cells with similar potency. Both sphingosine and C2­ceramide induce apoptosis in tumorigenic cells. Among the sphingosine stereoisomers (D­erythro, D­threo, L­erythro, and L­threo) and sphinganine that were tested, L­erythro­sphingosine most potently inhibits proliferation of tumorigenic cells. The inhibition of breast tumorigenic/cancer cell proliferation by sphingosine was accompanied by inhibition of telomerase activity. Sphingosine at non­cytotoxic concentrations, but not C2­ceramide, induces differentiation of normal stem cells (Type I), thereby reducing the number of stem cells that are more susceptible to neoplastic transformation. To the best of our knowledge, the present study demonstrates one of the first results that sphingosine can be a potential chemotherapeutic and cancer­protective agent, whereas C2­ceramide is not an ideal chemotherapeutic and cancer­protective agent due to its anti­proliferative effects on Type II HBECs and its inability to induce the differentiation of Type I to Type II HBECs.

A quantitative study of susceptibility and additional frequency shift of three common materials in MRI.

PURPOSE: This work quantifies magnetic susceptibilities and additional frequency shifts derived from different samples. METHODS: Twenty samples inside long straws were imaged with a multiecho susceptibility weighted imaging and analyzed with two approaches for comparisons. One approach applied our complex image summation around a spherical or cylindrical object method to phase distributions outside straws. The other approach utilized phase values inside each straw from two orientations. Both methods quantified susceptibilities of each sample at each echo time. The R2* value of each sample was measured too. Uncertainty of each measurement was also estimated. RESULTS: Quantified susceptibilities from complex image summation around a spherical or cylindrical object are consistent within uncertainties between different echo times. However, this is not the case for the other method. Nonetheless, most quantified susceptibilities are consistent between these two methods. Phase values due to additional frequency shifts in some of ferritin and nanoparticle samples have been identified. Only R2* values quantified from low concentration nanoparticle samples agree with the predictions from the static dephasing theory. CONCLUSION: This work suggests that using the sample sizes and phase values only outside samples can correctly quantify the susceptibilities of those samples. With the presence of a possible additional frequency shift inside a material, it will not be suitable to obtain susceptibility maps without taking that into account. Magn Reson Med 76:1263-1269, 2016. © 2015 Wiley Periodicals, Inc.

Susceptibility and size quantification of small human veins from an MRI method.

Recently a method called CISSCO (Complex Image Summation around a Spherical or a Cylindrical Object) was introduced for accurately quantifying the susceptibility and the radius of any narrow cylindrical object at any orientation using a typical two-echo gradient echo sequence. This work further optimizes the method for quantifying oxygen saturation in small cerebral veins in the human brain. The revised method is first validated through numerical simulations and then applied to data from phantom and human brain. The effect of phase high pass filtering on the quantified parameters is studied and procedures for mitigating its adverse effects are suggested. Uncertainty of each measurement is estimated from the error propagation method. It is shown that the revised method allows for accurate quantification of both the vessel size and its oxygen saturation even in the case of a low SNR (signal to noise ratio) in the vein. The results are self consistent across different veins within a given subject with a variation of less than 6%. Finally, imaging parameters and some procedures are suggested for accurate susceptibility and radius quantifications of small human veins.

An improved method for susceptibility and radius quantification of cylindrical objects from MRI.

A new method is developed to measure the magnetic susceptibilities and radii of small cylinder-like objects at arbitrary orientations accurately. This method for most biological substances only requires a standard gradient echo sequence with one or two echo times, depending on the orientation of an object relative to the main magnetic field. For objects oriented at the magic angle, however, this method is not applicable. As a byproduct of this method, the cross-sectional area as well as signals inside and outside the object can be determined. The uncertainty of each measurement is estimated from the error propagation method. Partial volume, dephasing, and phase aliasing effects are naturally included in the equations of this method. A number of simulations, phantom, and pilot in-vivo human studies are carried out to validate the theory. When the maximal phase value at the boundary of a given cylindrical object is larger than 3 radians, and the phase inside the object is more than 1 radian, the susceptibility can be accurately quantified within 15%. The radius of the object can be determined to subpixel accuracy. This is the case when the signal-to-noise ratio inside the object is about 6:1 or higher and the radius of the object is about one pixel or larger. These conditions are realistic when considering medullary and pial veins for example.

Magnetic moment quantifications of small spherical objects in MRI.

PURPOSE: The purpose of this work is to develop a method for accurately quantifying effective magnetic moments of spherical-like small objects from magnetic resonance imaging (MRI). A standard 3D gradient echo sequence with only one echo time is intended for our approach to measure the effective magnetic moment of a given object of interest. METHODS: Our method sums over complex MR signals around the object and equates those sums to equations derived from the magnetostatic theory. With those equations, our method is able to determine the center of the object with subpixel precision. By rewriting those equations, the effective magnetic moment of the object becomes the only unknown to be solved. Each quantified effective magnetic moment has an uncertainty that is derived from the error propagation method. If the volume of the object can be measured from spin echo images, the susceptibility difference between the object and its surrounding can be further quantified from the effective magnetic moment. Numerical simulations, a variety of glass beads in phantom studies with different MR imaging parameters from a 1.5T machine, and measurements from a SQUID (superconducting quantum interference device) based magnetometer have been conducted to test the robustness of our method. RESULTS: Quantified effective magnetic moments and susceptibility differences from different imaging parameters and methods all agree with each other within two standard deviations of estimated uncertainties. CONCLUSION: An MRI method is developed to accurately quantify the effective magnetic moment of a given small object of interest. Most results are accurate within 10% of true values, and roughly half of the total results are accurate within 5% of true values using very reasonable imaging parameters. Our method is minimally affected by the partial volume, dephasing, and phase aliasing effects. Our next goal is to apply this method to in vivo studies.

Evaluation of a commercial cardiac motion phantom for dual-energy chest radiography.

Misregistration due to cardiac motion causes artifacts in two-exposure dual-energy subtraction images, in both the soft-tissue-only image and the bone-only image. Two previous investigations have attempted to avoid misregistration artifacts by using cardiac gating of the first and second exposures. The severity of misregistration was affected by the heart rate, the time interval between the low- and high-energy exposures, the total duration of the two exposures, and the phase of the cardiac cycle at the start of the exposure sequence. We sought to determine whether a commercial phantom with a simulated beating heart can be used to investigate the factors affecting misregistration in dual-energy chest radiography. We made dual-energy images of the phantom in postero-anterior orientation using the indirect digital radiography system (GE XQ/i). We acquired digital images at heart rates between 40 beats per minute and 120 beats per minute and transferred them to a computer, where the area of the artifact on the silhouette of the heart was measured from both soft-tissue-only and bone-only images. For comparison, we measured misregistration in clinical dual-energy subtraction images by the same method. Generally speaking, without synchronization of the exposure sequence with the cardiac cycle, the area of the misregistration artifact increased with heart rate for both the phantom and clinical images. However, the phantom exaggerated the magnitude of misregistration relative to clinical images. Although this phantom was designed for horizontal operation and computed tomography imaging, it can be used in an upright configuration to simulate heart motion for investigation of dual-energy misregistration artifacts and control.

Quantifying effective magnetic moments of narrow cylindrical objects in MRI.

A new procedure for accurately measuring effective magnetic moments of long cylinders is presented. Partial volume, dephasing and phase aliasing effects are naturally included and overcome in our approach. Images from a typical gradient echo sequence at one single echo time are usually sufficient to quantify the effective magnetic moment of a cylindrical-like object. Only pixels in the neighborhood of the object are needed. Our approach can accurately quantify the magnetic moments and distinguish subpixel changes of cross sections between cylindrical objects. Uncertainties of our procedure are studied through the error propagation method. Images acquired with different parameters are used to test the robustness of our method. Alternate approaches and their limitations to extract magnet moments of objects with different orientations are also discussed. Our method has the potential to be applied to any long object whose cross section is close to a disk.

A complex sum method of quantifying susceptibilities in cylindrical objects: the first step toward quantitative diagnosis of small objects in MRI.

A complex sum method of quantifying the magnetic susceptibility of a long, narrow cylinder embedded in a uniform medium has been developed. The radius of the cylinder can be as small as one pixel. The susceptibility inside the object is extracted from the magnetic resonance complex images, using two concentric circles around the axis of the cylinder. The numerical simulations of this complex sum method are in good agreement with the phantom studies. Specifically, the method was tested with a susceptibility difference of -9 ppm to mimic air/tissue interface in the human body at 1.5 T with an echo time of 5 ms. Phantom studies using an air-filled cylinder in a solidified gel have shown that the susceptibility of the gel cannot be determined by the usual least-squares-fit method but can be determined by the complex sum method to within 5-10% of the expected value.

New approaches in identifying drugs to inactivate oncogene products.

With an information explosion on the molecular mechanism of oncogenesis, the completion of the human genome sequence project, and the advances in genomic and proteomic methods, many therapeutic targets for various cancers have been identified. It is timely that a number of new drug development techniques have been developed in this last decade. Candidate drug targets can now be efficiently validated with RNA interference and transgenic animals studies. Combinatorial chemistry provides large numbers of chemical compounds for drug lead discovery and optimization. High throughput assays and high content cell-based assays, in conjunction with sophisticated robotics, are now available for screening large numbers of compounds. Based on X-ray crystallographic structure data, drug leads can be discovered through in silico screening of virtual libraries. By applying these various drug discovery techniques, it is anticipated that more potent and specific anti-cancer agents will be discovered within the next decade.

The Tyr-kinase inhibitor AG879, that blocks the ETK-PAK1 interaction, suppresses the RAS-induced PAK1 activation and malignant transformation.

AG 879 has been widely used as a Tyr kinase inhibitor specific for ErbB2 and FLK-1, a VEGF receptor. The IC(50) for both ErbB2 and FLK-1 is around 1 microM. AG 879, in combination of PP1 (an inhibitor specific for Src kinase family), suppresses almost completely the growth of RAS-induced sarcomas in nude mice. In this paper we demonstrate that AG 879 even at 10 nM blocks the specific interaction between the Tyr-kinase ETK and PAK1 (a CDC42/ Rac-dependent Ser/Thr kinase) in cell culture. This interaction is essential for both the RAS-induced PAK1 activation and transformation of NIH 3T3 fibroblasts. However, AG 879 at 10 nM does not inhibit either the purified ETK or PAK1 directly in vitro, suggesting that this drug blocks the ETK-PAK1 pathway by targeting a highly sensitive kinase upstream of ETK. Although the Tyr-kinases Src and FAK are known to activate ETK directly, Src is insensitive to AG 879, and FAK is inhibited by 100 nM AG 879, but not by 10 nM AG879. The structure-function relationship analysis of AG 879 derivatives has revealed that both thio and tert-butyl groups of AG 879, but not (thio) amide group, are essential for its biological function (blocking the ETK-PAK1 pathway), suggesting that through the (thio) amide group, AG 879 can be covalently linked to agarose beads to form a bioactive affinity ligand useful for identifying the primary target of this drug.