Hemopoietic-specific Sf3b1-K700E knock-in mice display the splicing defect seen in human MDS but develop anemia without ring sideroblasts.

Heterozygous somatic mutations affecting the spliceosome gene SF3B1 drive age-related clonal hematopoiesis, myelodysplastic syndromes (MDS) and other neoplasms. To study their role in such disorders, we generated knock-in mice with hematopoietic-specific expression of Sf3b1-K700E, the commonest type of SF3B1 mutation in MDS. Sf3b1K700E/+ animals had impaired erythropoiesis and progressive anemia without ringed sideroblasts, as well as reduced hematopoietic stem cell numbers and host-repopulating fitness. To understand the molecular basis of these observations, we analyzed global RNA splicing in Sf3b1K700E/+ hematopoietic cells. Aberrant splicing was associated with the usage of cryptic 3' splice and branchpoint sites, as described for human SF3B1 mutants. However, we found a little overlap between aberrantly spliced mRNAs in mouse versus human, suggesting that anemia may be a consequence of globally disrupted splicing. Furthermore, the murine orthologues of genes associated with ring sideroblasts in human MDS, including Abcb7 and Tmem14c, were not aberrantly spliced in Sf3b1K700E/+ mice. Our findings demonstrate that, despite significant differences in affected transcripts, there is overlap in the phenotypes associated with SF3B1-K700E between human and mouse. Future studies should focus on understanding the basis of these similarities and differences as a means of deciphering the consequences of spliceosome gene mutations in MDS.

A DNA target-enrichment approach to detect mutations, copy number changes and immunoglobulin translocations in multiple myeloma.

Genomic lesions are not investigated during routine diagnostic workup for multiple myeloma (MM). Cytogenetic studies are performed to assess prognosis but with limited impact on therapeutic decisions. Recently, several recurrently mutated genes have been described, but their clinical value remains to be defined. Therefore, clinical-grade strategies to investigate the genomic landscape of myeloma samples are needed to integrate new and old prognostic markers. We developed a target-enrichment strategy followed by next-generation sequencing (NGS) to streamline simultaneous analysis of gene mutations, copy number changes and immunoglobulin heavy chain (IGH) translocations in MM in a high-throughput manner, and validated it in a panel of cell lines. We identified 548 likely oncogenic mutations in 182 genes. By integrating published data sets of NGS in MM, we retrieved a list of genes with significant relevance to myeloma and found that the mutational spectrum of primary samples and MM cell lines is partially overlapping. Gains and losses of chromosomes, chromosomal segments and gene loci were identified with accuracy comparable to conventional arrays, allowing identification of lesions with known prognostic significance. Furthermore, we identified IGH translocations with high positive and negative predictive value. Our approach could allow the identification of novel biomarkers with clinical relevance in myeloma.

Simultaneous localized 915 MHz external and interstitial microwave hyperthermia to heat tumors greater than 3 cm in depth.

A technique for heating superficial lesions extending to a depth greater than 3 cm using 915 MHz electromagnetic energy is described. It is well established that with external microwave applicators operating at 915 MHz, it is only possible to heat adequately up to a maximum depth of 3 cm. However, it is demonstrated that by implanting additional layers of interstitial microwave antennas (1.5 cm spacing) at required depths greater than 3 cm, and by simultaneously exciting these applicators as well as an external applicator, it is possible to extend the depth of heating at this frequency. A large neck node was successfully heated when this combined technique was used. The details of the method and the equipment used are described. Specific Absorption Rate (SAR) values and temperature distributions obtained during the four treatments delivered to the patient are also presented. This technique can be used in situations where it is not possible to perform extensive interstitial implantation, and in institutions where hyperthermia equipment for heating deeper lesions is not available.

Characteristics of improved microwave interstitial antennas for local hyperthermia.

The heating potentials of two newly-developed microwave interstitial antennas are reported in this paper. The longitudinal (parallel to the antenna) and transverse (over a plane perpendicular to the antenna) specific absorption rate (SAR) distributions of single and an array of four parallel antennas were measured in a muscle equivalent phantom and their performance characterized at 915 MHz in terms of the following parameters: peak depth (location of the profile peak with respect to the surface), 50% HL (effective heating length over which SAR greater than 50% of the peak normalized SAR), dead length (axial length at the antenna tip with SAR less than 50% of peak normalized SAR), and the variations of the specific absorption rate pattern relative to the depth of insertion. The results are analyzed and discussed in terms of these parameters and other factors important in the clinical use of these antennas for effective interstitial hyperthermia.

Adverse impact of multileaf collimator field shaping on lens dose in children with acute leukemia receiving cranial irradiation.

PURPOSE: This study was designed to investigate the impact of multileaf collimator (MLC) on lens dose in children with leukemia undergoing cranial irradiation. METHODS AND MATERIALS: This is a prospective study utilizing three common cranial irradiation techniques. Technique A uses a half-beam, nondivergent radiation field. Technique B has the anterior divergent field edge at the lateral bony canthus. Technique C is similar to B, but with a field collimator angle. Thermoluminescent dosimeter (TLD) lens dose measurements were obtained in children and phantom with all three techniques. RESULTS: Seventeen children were studied. Lens dose measurements were obtained in 14 children with technique A using MLC and blocks. In 7 of 14 children, dose measurements were obtained with MLC only. One child was treated with technique B and 2 children were treated with C, with MLC +/- blocks. In all 3 techniques, with MLC alone, the lens dose increased by 64%, 119%, and 72%, respectively. Similar results were obtained in phantom. CONCLUSION: This study demonstrates that independent of irradiation technique, additional custom blocking is required to maximally protect the lens with MLC shaped fields. This is due to the lack of conformity between MLC and the desired field edge at the lateral bony canthus.

Evaluation of an ingestible telemetric temperature sensor for deep hyperthermia applications.

We have investigated the potential of an ingestible thermometric system (ITS) for use with a deep heating system. The ingestible sensor contains a temperature-sensitive quartz crystal oscillator. The telemetered signal is inductively coupled by a radiofrequency coil system to an external receiver. The sensors, covered with a protective silicon coating, are 10 mm in diameter and 20 mm long and are energized by an internal silver-oxide battery. Experimental studies were carried out to investigate the accuracy of the system and the extent of reliable operation of these sensors in an electromagnetic environment. Different measurements were repeated for five sensors. Calibration accuracy was verified by comparison with a Bowman probe in the temperature range 30 degrees C to 55 degrees C. Linear regression analysis of individual pill readings indicated a correlation within +/- 0.4 degrees C at 95% prediction intervals in the clinical temperature range of 35 degrees C to 50 degrees C. Further work is required to improve this accuracy to meet the quality assurance guidelines of +/- 0.2 degrees C suggested by the Hyperthermia Physics Center. Response times were determined by the exponential fit of heat-up and cool-down curves for each pill. All curves had correlation coefficients greater than 0.98. Time (mean +/- SE) to achieve 90% response during heat-up was 115 +/- sec. Time to cool-down to 10% of initial temperature was 114 +/- 4 sec. The effect of the external antenna and sensor spacing and the angle of orientation of the sensor relative to the antenna plane were also studied. Electromagnetic interference effects were studied by placing the sensor with a Bowman probe in a cylindrical saline phantom for the tests in an annular phase array applicator. Different power levels at three frequencies--80, 100, and 120 MHz--were used. Accurate temperature readings could not be obtained when the electromagnetic power was on because of interference effects with the receiver. However, the temperatures read with the ITS immediately after the electromagnetic power was switched off correlated well with the Bowman probe readings across the power categories and the three frequencies used. The phantom was heated to steady state, with a Bowman probe placed at the central axis of the cylinder used as control. During the heat-up period and the steady state, the mean difference (+/- SE) between the ITS and Bowman probe was 0.12 degrees C (+/- 0.05 degrees C).(ABSTRACT TRUNCATED AT 400 WORDS)

Initial results for automated computational modeling of patient-specific electromagnetic hyperthermia.

Developments in finite-difference time-domain (FD-TD) computational modeling of Maxwell's equations, super-computer technology, and computed tomography (CT) imagery open the possibility of accurate numerical simulation of electromagnetic (EM) wave interactions with specific, complex, biological tissue structures. One application of this technology is in the area of treatment planning for EM hyperthermia. In this paper, we report the first highly automated CT image segmentation and interpolation scheme applied to model patient-specific EM hyperthermia. This novel system is based on sophisticated tools from the artificial intelligence, computer vision, and computer graphics disciplines. It permits CT-based patient-specific hyperthermia models to be constructed without tedious manual contouring on digitizing pads or CRT screens. The system permits in principle near real-time assistance in hyperthermia treatment planning. We apply this system to interpret actual patient CT data, reconstructing a 3-D model of the human thigh from a collection of 29 serial CT images at 10 mm intervals. Then, using FD-TD, we obtain 2-D and 3-D models of EM hyperthermia of this thigh due to a waveguide applicator. We find that different results are obtained from the 2-D and 3-D models, and conclude that full 3-D tissue models are required for future clinical usage.

Re-irradiation and external hyperthermia in locally advanced, radiation recurrent, hormone refractory prostate cancer: a preliminary report.

The purpose of this report is to present the preliminary results of re-irradiation and external hyperthermia in patients with locally advanced, previously irradiated, hormone refractory prostate cancer. Three consecutive patients with symptomatic, locally advanced, previously irradiated and hormone refractory prostate cancer were treated with further irradiation (30.6-50 Gy) and external hyperthermia (5-8 treatments). All patients had complete resolution of symptoms lasting for 12-24 months. Significant tumour shrinkage, including complete tumour response, was demonstrated by CT and endoscopy. In one case, at 2 years after re-treatment, there is continued tumour regression and bone regeneration in the pelvis. Two patients had local control of tumour, which continued until most recent follow-up at 12 months and more than 24 months, respectively. Another case developed local recurrence at 17 months. At most recent follow-up, no patient has experienced significant treatment-related side effects. In these patients with no other therapeutic alternatives, re-irradiation and hyperthermia can provide durable tumour response for more than a year, resulting in significant improvement in quality of life. Further clinical studies are warranted.

A clinical microwave hyperthermia system with multipoint real-time thermal dosimetry.

A clinical hyperthermia system using a 915 MHz microwave generator and incorporating multipoint thermocouple thermometry is described. Temperatures can be monitored simultaneously at 16 points and measurements displayed on a visual display unit and a plotter. The power output of the generator is adjusted under computer control to maintain a constant predetermined temperature in a chosen control channel. A clinically useful feature of the system is the ability to determine the effective cumulative thermal dose delivered to the tissue at points monitored in real time. The basis for the thermal dose calculation is discussed in detail. The calculated dose parameter is displayed for each of the 16 channels during the treatment and updated every 30 s. This real-time display of a thermal dose parameter has made possible a more homogeneous heating of tumours.

Regional hyperthermia for extensive pelvic tumours using an annular phased array applicator: a feasibility study.

Twenty patients with extensive pelvic tumours have been treated with radiotherapy and regional hyperthermia using an annular phased array applicator (APA). The majority of patients had bulky disease with a mean tumour volume of 206 cm3. In 63 of 67 attempted treatments heating was achieved, temperatures in excess of 42 degrees C being reached in the pelvis in 78%. Heat doses, however, were low, only 41% of treatments exceeding 5 min equivalent at 43 degrees C. Treatment was usually limited by acute toxicity and, although this was marked, it was of short duration and no long-term sequelae have been noted. With gain in operator experience, later treatments have achieved improved thermal doses. Response was difficult to assess but we have documented responses in six of 12 assessable patients. Palliation has been achieved in all patients with symptoms. Preliminary attempts to shift the region of maximum heating by phase variations in the APA output are promising, and this technique should improve the quality of treatments and reduce toxicity. The APA is thus capable of inducing pelvic regional hyperthermia, and a randomised controlled study is now planned to assess the value of pelvic hyperthermia as a treatment modality.

SU-E-T-570: Improvement to the Histogram Analysis in Radiation Therapy (HART): An Open Source Software System for the Multi-Dimensional Dose- Volume Histogram Analysis in Digital Image Communication in Medicine - Radiation Therapy (DICOM-RT) Treatment Plans.

PURPOSE: Histogram Analysis in Radiation Therapy (HART) is an efficient and accurate dose-volume histogram (DVH) computational tool in radiotherapy research. Several applications of the program have been presented previously (J Appl Clin Med Phys 11(1): 3013, 2010; Med Phys 38(6), p.3678, 2011) for the Radiation Therapy Oncology Group (RTOG) users. The program has been further developed to incorporate various types of DVH analysis features to support the research using DICOM-RT plans. The main objective of this work was to present the improvement and compatibility of the program for the DICOM-RT plans. METHODS AND MATERIALS: MATLAB based codes were primarily designed to read and write a simpler HART format from the standard DICOM-RT data objects exported from the Xio treatment planning system (CMS Inc., St. Louis, MO). This format employed an optimal polynomial fitting technique to interpolate the co-ordinates of the contours in the regions-of-interest. The format was efficient for the (a) precise extraction of the cumulative DVH (cDVH) and spatial DVH (sDVH; x-,y-, and z-DVHs respectively) data- statistics, (b) universal-plan indices evaluation, (c) biological modeling based outcome analyses (BMOA), (d) radiobiological dose-response modeling, and (e) physical parameterization modules. The fundamental DVH statistics were validated using the DVH statistics extracted from the Computational Environment for Radiotherapy Research program. RESULTS: HART offers various types of DVH computational functionalities, several plan evaluation and radiobiological outcome analysis modules in a user- friendly software package for the RTOG and DICOM-RT planners. The cDVH and BMOA modules were found to be the most applicable features for the global researchers. CONCLUSIONS: HART is a novel and universal multi-dimensional DVH analysis tool for the radiation therapy research. We further expect to develop HART for the space-time DVH analysis and proton therapy applications. The software is available online (http://www2.uic.edu/∼apyaku1) for the radiotherapy research. This work was partially supported by NIH-NIDCD grant.

Hyperthermia and radiation in the treatment of superficial malignancy: an analysis of treatment parameters, response and toxicity.

A total of 41 superficial tumours have been treated with radiation alone (21 lesions) or in combination with hyperthermia (20 lesions) in a non-randomized study on 16 patients. A minimum tumour heat dose in excess of 30 min equivalent at 43 degrees C was achieved in 46 of the 80 (57 per cent) hyperthermia treatments. Small lesions received better quality heat treatments and were more likely to achieve a complete response. There is a significant increase in the response of lesions treated with the combined modality compared with radiation alone (P less than 0.01). Similar results were obtained when matched lesions with internal controls were analysed separately. There was an increased incidence of severe skin reactions in the hyperthermia-treated group with the reaction tending to develop more quickly. There were three instances of late fibrosis in the hyperthermia group. There is a significant correlation between the severity of the skin reaction and the average maximum skin heat dose per treatment.

Dual-antenna applicator for hyperthermia of tumours at intermediate depth.

A dual-antenna applicator with 21 x 26 cm2 aperture, that is fully loaded and operates at 74 MHz, was developed at the Mallinckrodt Institute of Radiology. By placing two antennas into an applicator capable of propagating TE10 mode, a significant enlargement of heating pattern was achieved without an increase in applicator dimensions. When antennas are placed symmetrically about a parallel to the antenna axis of symmetry, the sensitivity of the applicator input impedance to variations of load impedance reduces. Stable coupling of the RF power to the treatment area may be provided. Twenty patients with eccentrically located tumours were treated using this device.

Potential for localized, adjustable deep heating in soft-tissue environments with a 30-beam ultrasonic hyperthermia system.

Initial heating rates (degrees C/min) along parallel tracks at depths of 1-14 cm in a static, muscle-like phantom were determined from time-temperature profiles obtained with 'Helios', a 30-beam ultrasonic hyperthermia system developed by Varian Associates. Data were taken at a single operating frequency of 556 kHz, for different sets of focal plane ring diameters of the four-ring array applicator, different levels of transducer driving power and two different focal plane depths, 6 cm and 9 cm. In each experiment, at each point of temperature measurement, analysis of temperature versus time data over a 2 min heating interval permitted separation of the desired phantom heating from artefactual heating resulting primarily from absorption of transverse (shear) waves produced at phantom-metal probe catheter interfaces. The results of the studies conducted suggest that in a non-translating carriage mode, Helios can produce axially and laterally localized deep heating in soft tissues for tissue volumes of lateral dimension up to a minimum of 4 cm and tissue depths of at least 11 cm. The results obtained also suggest that Helios can produce laterally localized heating to tissue depths of at least 11 cm without excessive heating of superficial soft-tissue layers, for tissue volumes of lateral dimension up to a minimum of 8 cm. The methodology used in the phantom studies was applied to the production of localized heating in the right lobe of the liver of adult pigs. Temperature versus time profiles obtained in the in vivo studies indicated that, for the set of system parameters employed, concentration of ultrasonic power at greater depths in the liver (e.g. 10.5 cm versus 5 cm) could be achieved, suggesting that Helios should be able to produce localized heating of targeted hepatic volumes when its operating parameters are selected in accordance with effective treatment planning techniques.

Regional hyperthermia in the treatment of recurrent deep-seated tumors: preliminary analysis of phase I trial.

This is a preliminary report on a phase I pilot study of regional hyperthermia of advanced tumors, using annular phased array system (APAS) in combination with radiation therapy and/or chemotherapy. All patients were treated without anesthesia. In all hyperthermia sessions, physiological parameters such as blood pressure, cardiac rate, respiratory rate, and EKG were monitored throughout the treatment. Tumor temperature was monitored with intratumoral probe with an additional probe either in the esophagus or rectum for monitoring systemic temperature. Present reports are on analysis of the first 30 patients: a total of 139 treatment sessions were analyzed. The results indicate local discomfort, anxiety, systemic temperature elevation, and tachycardia were the predominant factors limiting the number of sessions, the duration of sessions, and the deposition of power, in 18, 6, 6, and 2 patients, respectively. The complications were rare. No injuries to visceral organs clearly attributed to the hyperthermia have been recorded.

Physical predictors of adequate hyperthermia with the annular phased array.

One hundred and fifty-two hyperthermia sessions in 36 consecutive patients treated with the BSD 1000 annular phased array system (APAS) are reviewed with regard to physical predictors of quality of hyperthermia. Although central tumour temperatures exceeding 42 degrees C were momentarily obtained in 62% of the sessions, it frequently proved difficult to maintain the patient at temperature for prolonged periods of time. The time to reach target temperature was negatively associated with quality of hyperthermia. Thus, of 25 sessions which required over 25 min to attain a temperature of 42 degrees C, only one was adequate (defined as central tumour temperature greater than or equal to 42 degrees C maintained for at least 30 min) as opposed to 28/69 adequate sessions when 42 degrees C was reached in less than 25 min. Physical parameters measured in the first 3 min of the session found to be associated with adequate hyperthermia include an initial rate of temperature rise at the tumour site exceeding 0.4 degrees C/min achieved with a net forward power less than 1500 W. Only three of 57 sessions not meeting these criteria were adequate. Treatment policy recommendations and recommendations for future research are made.

Effects of hyperthermia and iodine-131-labeled anticarcinoembryonic antigen monoclonal antibody on human tumor xenografts in nude mice.

BACKGROUND: Many studies have demonstrated synergistic interaction between hyperthermia and radiation. This study was undertaken to determine whether hyperthermia could enhance the effect of radioimmunotherapy (RIT) in the treatment of human colon adenocarcinoma xenografts in nude mice. METHODS: The experiments were conducted in two parts. During the first part of the study, preliminary information was obtained regarding the effect of various temperatures (41 degrees C, 42 degrees C, and 43 degrees C for 45 minutes) and iodine-131-labeled anticarcinoembryonic antigen (CEA) monoclonal antibodies (RMoAb) with administered activity ranging from 130 +/- 19 microCi to 546 +/- 19 microCi on tumor regrowth delay (TRD) and volume doubling time. This information was used in Part 2 of the study, which included four groups of mice: (1) a control group, (2) a group treated with hyperthermia, (3) a group treated with RMoAb, and (4) a group treated with a combination of RMoAb and hyperthermia. RESULTS: Maximum and significantly increased TRD was observed in the group treated with RMoAb and hyperthermia (slope, 0.057) compared with the control group (slope, 0.322), the hyperthermia-treated group (slope, 0.302), and the group treated with RMoAb alone (slope, 0.098). The ratio of the slopes between the groups treated with RMoAb and those treated with RMoAb and hyperthermia was 1.72. No correlation was detected between the percent of antibody uptake in the tumor and tumor regression in the groups treated with heat and RMoAb and those treated with RMoAb alone. CONCLUSIONS: The results of these experiments show that hyperthermia increased the effectiveness of iodine-131-labeled anti-CEA monoclonal antibodies against human colon carcinoma xenografts in nude mice. This study offers a rationale for combining hyperthermia and low-dose radiation produced from RIT in clinical practice.

Phase I/II trial of combined 131I anti-CEA monoclonal antibody and hyperthermia in patients with advanced colorectal adenocarcinoma.

BACKGROUND: This pilot project was undertaken to evaluate the toxicity of and tumor response to combined 131I anti-carcinoembryonic antigen monoclonal antibody (131I anti-CEA RMoAb) and hyperthermia in patients with metastatic colorectal adenocarcinoma. METHODS: Nine patients who had colorectal carcinoma with liver metastases were enrolled in this study. Intact 131I anti-CEA RMoAb was used (the specific antibody was IMMU-4, provided by Immunomedics, Inc., Morris Plains, NJ). During the diagnostic phase, dosimetry revealed that the tumor site received a higher radiation dose than the surrounding normal tissues in only six patients. These six, who were treated with radioimmunotherapy and hyperthermia, were the basis of this study. The first three patients were treated with 30 mCi/m2 of 131I anti-CEA RMoAb, and the next three received 60 mCi/m2. Pharmacokinetic clearance data were reported for all nine patients. RESULTS: Thermometry data revealed an average T90 of 40.3 (+/- 1.4 degrees C) and T50 of 41.1 (+/- 1.2 degrees C). The average thermal dose equivalent at 42.5 degrees C was 34.5 (+/- 21.5) minutes. The average Tmin, Tmax, and Tmeam were 40 (+/- 1.2 degrees C), 42.4 (+/- 0.7 degrees C), and 41.1 (+/- 1.1 degrees C), respectively. The pharmacokinetic clearance data of antibody showed monoexponential plasma clearances in all patients except one, in whom a biexponential plasma clearance was observed. In general, similar plasma and whole-body clearances as well as similar urinary excretions were observed when diagnostic and therapeutic phases for each patient were compared. Two of the six patients showed a marked improvement in their symptoms; five patients showed a drop in carcinoembryonic antigen levels. A follow-up computed tomography scan one month after treatment showed no change in tumor volume in five patients; one patient showed a partial response. Three patients developed toxicity, two developed moderate thrombocytopenia (39,000 and 58,000), and the other patient developed hematoma resulting from the insertion of a catheter for thermometry. CONCLUSIONS: It is feasible to combine hyperthermia and radiolabeled monoclonal antibodies, and the combination was well tolerated by these patients. The interaction between hyperthermia and low dose rate radioimmunotherapy is complex. Further studies are necessary to explore the use of this combined modality in the management of maligancies.