Psoralen as a Photosensitizers for Photodynamic Therapy by Means of In Vitro Cherenkov Light.

Possible enhancements of DNA damage with light of different wavelengths and ionizing radiation (Rhenium-188-a high energy beta emitter (Re-188)) on plasmid DNA and FaDu cells via psoralen were investigated. The biophysical experimental setup could also be used to investigate additional DNA damage due to photodynamic effects, resulting from Cherenkov light. Conformational changes of plasmid DNA due to DNA damage were detected and quantified by gel electrophoresis and fluorescent staining. The clonogene survival of the FaDu cells was analyzed with colony formation assays. Dimethyl sulfoxide was chosen as a chemical modulator, and Re-188 was used to evaluate the radiotoxicity and light (UVC: λ = 254 nm and UVA: λ = 366 nm) to determine the phototoxicity. Psoralen did not show chemotoxic effects on the plasmid DNA or FaDu cells. After additional treatment with light (only 366 nm-not seen with 254 nm), a concentration-dependent increase in single strand breaks (SSBs) was visible, resulting in a decrease in the survival fraction due to the photochemical activation of psoralen. Whilst UVC light was phototoxic, UVA light did not conclude in DNA strand breaks. Re-188 showed typical radiotoxic effects with SSBs, double strand breaks, and an overall reduced cell survival for both the plasmid DNA and FaDu cells. While psoralen and UVA light showed an increased toxicity on plasmid DNA and human cancer cells, Re-188, in combination with psoralen, did not provoke additional DNA damage via Cherenkov light.

Radioimmunotherapy in Combination with Reduced-Intensity Conditioning for Allogeneic Hematopoietic Cell Transplantation in Patients with Advanced Multiple Myeloma.

Radioimmunotherapy (RIT) has the potential to reduce the incidence of relapse after allogeneic hematopoietic cell transplantation (allo-HCT) in patients with advanced-stage multiple myeloma (MM). In this study, we evaluated the efficacy of RIT in combination with chemotherapy-based reduced-intensity conditioning (RIC). RIT was based on the coupling of an anti-CD66 antibody to the beta emitter 188-rhenium (188-re) for targeted bone marrow irradiation. Between 2012 and 2018, 30 patients with MM, most of them heavily pretreated with various therapies including proteasome inhibitors, immunomodulatory drugs, anti-CD38 antibodies, and autologous hematopoietic cell transplantation (auto-HCT), were treated with a RIT-RIC combination before allo-HCT. In addition to a fludarabine plus melphalan- or treosulfan-based RIC, a median dose of 18.1 Gy (interquartile range [IQR], 14.6 to 24.1 Gy) was applied to the bone marrow. After a median duration of follow-up for surviving patients of 2.1 years (IQR, 1.3 to 3.0 years), the 2-year progression-free survival and overall survival rates were 43% (95% confidence interval [CI], 26% to 73%) and 55% (95% CI, 38% to 79%), respectively. The 2-year nonrelapse mortality and cumulative incidence of progression were 17% (95% CI, 3% to 30%) and 46% (95% CI, 25% to 67%), respectively. Renal toxicity and mucositis were the most frequent extramedullary side effects. In conclusion, the addition of RIT to RIC was safe and feasible and resulted in promising outcomes compared with those previously reported for RIC-based allo-HCT without the addition of RIT in patients with relapsed/refractory MM. Nevertheless, despite the addition of RIT, relapse after allo-HCT remained a major determinant of therapeutic failure. Therefore, the development of novel RIT strategies (eg, dual targeting strategies or combinations with adapter chimeric antigen receptor T cell-based therapies) is needed.

Twins in spirit part II: DOTATATE and high-affinity DOTATATE--the clinical experience.

PURPOSE: Over recent decades interest in diagnosis and treatment of neuroendocrine tumours (NET) has steadily grown. The basis for diagnosis and therapy of NET with radiolabelled somatostatin (hsst) analogues is the variable overexpression of hsst receptors (hsst1-5 receptors). We hypothesized that radiometal derivatives of DOTA-iodo-Tyr(3)-octreotide analogues might be excellent candidates for somatostatin receptor imaging. We therefore explored the diagnostic potential of (68)Ga-DOTA-iodo-Tyr(3)-octreotate [(68)Ga-DOTA,3-iodo-Tyr(3),Thr(8)]octreotide ((68)Ga-HA-DOTATATE; HA, high-affinity) compared to the established (68)Ga-DOTA-Tyr(3)-octreotate ((68)Ga-DOTATATE) in vivo. METHODS: The study included 23 patients with known somatostatin receptor-positive metastases from NETs, thyroid cancer or glomus tumours who were investigated with both (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE. A patient-based and a lesion-based comparative analysis was carried out of normal tissue distribution and lesion detectability in a qualitative and a semiquantitative manner. RESULTS: (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE showed comparable uptake in the liver (SUVmean 8.9 ± 2.2 vs. 9.3 ± 2.5, n.s.), renal cortex (SUVmean 13.3 ± 3.9 vs. 14.5 ± 3.7, n.s.) and spleen (SUVmean 24.0 ± 6.7 vs. 22.9 ± 7.3, n.s.). A somewhat higher pituitary uptake was found with (68)Ga-HA-DOTATATE (SUVmean 6.3 ± 1.8 vs. 5.4 ± 2.1, p < 0.05). On a lesion-by-lesion basis a total of 344 lesions were detected. (68)Ga-HA-DOTATATE demonstrated 328 lesions (95.3% of total lesions seen), and (68)Ga-DOTATATE demonstrated 332 lesions (96.4%). The mean SUVmax of all lesions was not significantly different between (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE (17.8 ± 11.4 vs. 16.7 ± 10.7, n.s.). CONCLUSION: Our analysis demonstrated very good concordance between (68)Ga-HA-DOTATATE and (68)Ga-DOTATATE PET data. As the availability and use of (68)Ga-HA-DOTATATE is not governed by patent restrictions it may be an attractive alternative to other (68)Ga-labelled hsst analogues.

Gamma camera imaging characteristics of 166Ho and 99mTc used in Selective Internal Radiation Therapy.

BACKGROUND: The administration of a 166Ho scout dose is available as an alternative to 99mTc particles for pre-treatment imaging in Selective Internal Radiation Therapy (SIRT). It has been reported that the 166Ho scout dose may be more accurate for the prediction of microsphere distribution and the associated therapy planning. The aim of the current study is to compare the scintigraphic imaging characteristics of both isotopes, considering the objectives of the pre-treatment imaging using clinically geared phantoms. METHODS: Planar and SPECT/CT images were obtained using a NEMA image quality phantom in different phantom setups and another body-shaped phantom with several inserts. The influence of collimator type, count statistics, dead time effects, isotope properties and patient obesity on spatial resolution, contrast recovery and the detectability of small activity accumulations was investigated. Furthermore, the effects of the imaging characteristics on personalized dosimetry are discussed. RESULTS: The images with 99mTc showed up to 3 mm better spatial resolution, up to two times higher contrast recovery and significantly lower image noise than those with 166Ho. The contrast-to-noise ratio was up to five times higher for 99mTc than for 166Ho. Only when using 99mTc all activity-filled spheres could be distinguished from the activity-filled background. The measurements mimicking an obese patient resulted in a degraded image quality for both isotopes. CONCLUSIONS: Our measurements demonstrate better scintigraphic imaging properties for 99mTc compared to 166Ho in terms of spatial resolution, contrast recovery, image noise, and lesion detectability. While the 166Ho scout dose promises better prediction of the microsphere distribution, it is important to consider the inferior imaging characteristics of 166Ho, which may affect individualized treatment planning in SIRT.

Alpha-Emitting Radionuclides: Current Status and Future Perspectives.

The use of radionuclides for targeted endoradiotherapy is a rapidly growing field in oncology. In particular, the focus on the biological effects of different radiation qualities is an important factor in understanding and implementing new therapies. Together with the combined approach of imaging and therapy, therapeutic nuclear medicine has recently made great progress. A particular area of research is the use of alpha-emitting radionuclides, which have unique physical properties associated with outstanding advantages, e.g., for single tumor cell targeting. Here, recent results and open questions regarding the production of alpha-emitting isotopes as well as their chemical combination with carrier molecules and clinical experience from compassionate use reports and clinical trials are discussed.

Investigation of Photodynamic Therapy Promoted by Cherenkov Light Activated Photosensitizers-New Aspects and Revelations.

This work investigates the proposed enhanced efficacy of photodynamic therapy (PDT) by activating photosensitizers (PSs) with Cherenkov light (CL). The approaches of Yoon et al. to test the effect of CL with external radiation were taken up and refined. The results were used to transfer the applied scheme from external radiation therapy to radionuclide therapy in nuclear medicine. Here, the CL for the activation of the PSs (psoralen and trioxsalen) is generated by the ionizing radiation from rhenium-188 (a high-energy beta-emitter, Re-188). In vitro cell survival studies were performed on FaDu, B16 and 4T1 cells. A characterization of the PSs (absorbance measurement and gel electrophoresis) and the CL produced by Re-188 (luminescence measurement) was performed as well as a comparison of clonogenic assays with and without PSs. The methods of Yoon et al. were reproduced with a beam line at our facility to validate their results. In our studies with different concentrations of PS and considering the negative controls without PS, the statements of Yoon et al. regarding the positive effect of CL could not be confirmed. There are slight differences in survival fractions, but they are not significant when considering the differences in the controls. Gel electrophoresis showed a dominance of trioxsalen over psoralen in conclusion of single and double strand breaks in plasmid DNA, suggesting a superiority of trioxsalen as a PS (when irradiated with UVA). In addition, absorption measurements showed that these PSs do not need to be shielded from ambient light during the experiment. An observational test setup for a PDT nuclear medicine approach was found. The CL spectrum of Re-188 was measured. Fluctuating inconclusive results from clonogenic assays were found.

18F-Choline, 11C-choline and 11C-acetate PET/CT: comparative analysis for imaging prostate cancer patients.

Prostate cancer (PCA) is the second most common tumour in men worldwide. Whereas prostate specific antigen (PSA) is an established biochemical marker, the optimal imaging method for all clinical scenarios has not yet been found. With the rising number of PET centres there is an increasing availability and use of (18)F-/(11)C-choline or (11)C-acetate for staging of PCA. However, to date no final conclusion has been reached as to whether acetate or choline tracers should be preferred. In this review we provide an overview of the performance of choline and acetate PET for staging the primary and recurrent disease and lymph nodes in PCA, based on the literature of the last 10 years. Although predominantly choline has been used rather than acetate, both tracers performed in a similar manner in published studies. Choline as well as acetate have insufficient diagnostic accuracy for the staging of the primary tumour, due to a minimum detectable tumour size of 5 mm and inability to differentiate PCA from benign prostate hyperplasia, chronic prostatitis and high-grade intraepithelial neoplasia. Regarding lymph node staging, choline tracers have demonstrated a high specificity. Unfortunately, the sensitivity is only moderate. For staging recurrent disease, sensitivity depends on the level of serum PSA (PSA should be >2 ng/ml). This applies to both choline and acetate. However, despite these limitations, a significant number of patients with recurrent disease can benefit from PET imaging by a change in treatment planning.

Up-Regulation of PSMA Expression In Vitro as Potential Application in Prostate Cancer Therapy.

Possibilities to improve the therapeutic efficacy of Lu-177-PSMA-617 radionuclide therapy by modulation of target expression are being investigated. Knowledge on regulatory factors that promote prostate cancer (PCa) progression may contribute to targeting prostate cancer more effectively. We aimed at the stimulation of PCa cell lines using the substances 5-aza-2'-deoxycitidine (5-aza-dC) and valproic acid (VPA) to achieve increased prostate-specific membrane antigen (PSMA) expression. PC3, PC3-PSMA, and LNCaP cells were incubated with varying concentrations of 5-aza-dC and VPA to investigate the cell-bound activity of Lu-177-PSMA-617. Stimulation effects on both the genetically modified cell line PC3-PSMA and the endogenously PSMA-expressing LNCaP cells were demonstrated by increased cellular uptake of the radioligand. For PC3-PSMA cells, the fraction of cell-bound radioactivity was enhanced by about 20-fold compared to that of the unstimulated cells. Our study reveals an increased radioligand uptake mediated by stimulation for both PC3-PSMA and LNCaP cell lines. In perspective of an enhanced PSMA expression, the present study might contribute to advanced radionuclide therapy approaches that improve the therapeutic efficacy, as well as combined treatment options.

Determination of Radiation Exposure of Individuals in the Population by Patients after Radioiodine Therapy - Comparison of two Measurement Systems. / Ermittlung der Strahlenexposition von Einzelpersonen in der Bevölkerung durch Patienten nach einer Radiojodtherapie ­ Vergleich von 2 Messsystemen.

According to the requirements of radiation protection legislation, patients may only be discharged from the nuclear medicine therapy ward if it is ensured that the cumulative radiation exposure of the population is below 1 mSv per year. In the present study, dose measurements of patients after radioiodine therapy (RIT) and their relatives are to be used to prove that the radiation exposure resulting from the medical application is low and that the legal framework conditions are complied with. Furthermore, the results allow conclusions to be drawn about the measurement accuracy of the dosimeters used. METHODS: In 147 patients after RIT and their relatives, the dosage was measured over 14 days with different measuring systems. Finger ring dosimeters (FRD) were worn during the whole day, furthermore the dose was determined by non-official OSL and TLD dosimeters during the sleep phase. RESULTS: 88 data sets were used for the final analysis. With the FRD, dose values between 0.1-50 mSv were determined for the patients. As expected, the finger ring dose of the relatives was significantly lower, averaging 0.75 mSv compared to 10 mSv for the patient. For the TLD and OSL used in the sleep phase, the measured values were in the same range. The reproducibility of the measurement results was significantly better for the OSL than for the TLD. CONCLUSION: Despite method-related measurement uncertainties, it can be concluded that the exposure dose of patients' relatives after radioiodine therapy is low and that the legal requirements are met. Moreover, the now official OSL dosimeters represent a more accurate and for the chosen measurement task better suited measurement system than the TLD. KEY POINTS: · The exposure dose of patients' relatives after radioiodine therapy is low.. · The requirements of radiation protection legislation after discharge from the nuclear medicine therapy ward are complied with. · OSL dosimeters are a accurate and for the measurement task suited system. CITATION FORMAT: · Hartmann H, Andreeff M, Claußnitzer J et al. Determination of Radiation Exposure of Individuals in the Population by Patients after Radioiodine Therapy - Comparison of two Measurement Systems. Fortschr Röntgenstr 2023; 195: 605 - 612.

Combining Cisplatin with Different Radiation Qualities-Interpretation of Cytotoxic Effects In Vitro by Isobolographic Analysis.

BACKGROUND: The combination of platinum-containing cytostatic drugs with different radiation qualities has been studied for years. Despite their massive side effects, these drugs still belong to the therapeutic portfolio in cancer treatment. To overcome the disadvantages of cisplatin, our study investigated the cytotoxic effects of combining radionuclides with cisplatin. METHODS: FaDu cells were treated with cisplatin (concentration ≈ 2 µM) and additionally irradiated after two hours with the alpha-emitter 223Ra, the beta-emitter 188Re as well as external X-rays using dose ranges of 2-6 Gy. Cell survival was followed by colony formation assays and plotted against cisplatin concentration and radiation dose. The results were interpreted by isobolograms. RESULTS: Isobolographic analyses revealed a supra-additive cytotoxic effect for the combination of cisplatin and 223Ra. A sub-additive effect was observed for the combination of cisplatin and 188Re, whereas a protective effect was found for the combination with X-rays. CONCLUSIONS: The combination of cisplatin and 223Ra may have the potential to create a successfully working therapy scheme for various therapy approaches, whereas the combination with 188Re as well as single-dose X-ray treatment did not lead to a detectable radiosensitizing effect. Thus, the combination with alpha-emitters might be advantageous and, therefore, should be followed in future studies when combined with cytostatic drugs.

Successful Combination of Olaparib and 225Ac-Dotatate in a Patient with Neuroendocrine Tumor G3 and BRCA Mutation.

Based on the results of the NETTER-1 trial, peptide receptor radionuclide therapy with Lutetium-177 (177Lu) - DOTATATE is authorized for the treatment of neuroendocrine tumors (NET) grade 1 (G1) and grade 2 (G2) of the intestine. After the failure of 177Lu-DOTATATE therapy, targeted alpha-particle therapy (TAT) may be a possible treatment option. Here, we present a patient with cancer of unknown primary NET G2 later G3. The patient was referred to our hospital with urosepsis due to a second-degree urinary retention. After stent insertion, a contrast-enhanced computed tomography revealed a huge pelvic tumor without metastases. Initially, the patient had undergone surgical treatment. Later the patient developed liver metastasis and was treated by 177Lu-DOTATATE therapy and four lines of systemic therapy. A disease progression was observed and with the knowledge of a germline BRCA1 mutation, the patient was treated with TAT (Actinium-225 [225Ac]-DOTATATE) combined with olaparib. The patient achieved a significant treatment response for 12 months indicating that a combination therapy with an alpha emitter and olaparib demands further investigations in clinical trials.

Epigenetic-Like Stimulation of Receptor Expression in SSTR2 Transfected HEK293 Cells as a New Therapeutic Strategy.

The aim of the study was to increase the uptake of the SSTR2-targeted radioligand Lu-177-DOTATATE using the DNA methyltransferase inhibitor (DNMTi) 5-aza-2'-deoxycytidine (5-aza-dC) and the histone deacetylase inhibitor (HDACi) valproic acid (VPA). The HEKsst2 and PC3 cells were incubated with variable concentrations of 5-aza-dC and VPA to investigate the uptake of Lu-177-DOTATATE. Cell survival, subsequent to external X-rays (0.6 or 1.2 Gy) and a 24 h incubation with 57.5 or 136 kBq/mL Lu-177-DOTATATE, was investigated via colony formation assay to examine the effect of the epidrugs. In the case of stimulated HEKsst2 cells, the uptake of Lu-177-DOTATATE increased by a factor of 28 in comparison to the unstimulated cells. Further, stimulated HEKsst2 cells demonstrated lower survival fractions (factor 4). The survival fractions of the PC3 cells remained almost unchanged. VPA and 5-aza-dC did not induce changes to the intrinsic radiosensitivity of the cells after X-ray irradiation. Clear stimulatory effects on HEKsst2 cells were demonstrated by increased cell uptake of the radioligand and enhanced SST2 receptor quantity. In conclusion, the investigated approach is suitable to stimulate the somatostatin receptor expression and thus the uptake of Lu-177-DOTATATE, enabling a more efficient treatment for patients with poor response to peptide radionuclide therapy (PRRT).

Ac-EAZY! Towards GMP-Compliant Module Syntheses of 225Ac-Labeled Peptides for Clinical Application.

The application of 225Ac (half-life T1/2 = 9.92 d) dramatically reduces the activity used for peptide receptor radionuclide therapy by a factor of 1000 in comparison to 90Y, 177Lu or 188Re while maintaining the therapeutic outcome. Additionally, the range of alpha particles of 225Ac and its daughter nuclides in tissue is much lower (47-85 µm for alpha energies Eα = 5.8-8.4 MeV), which results in a very precise dose deposition within the tumor. DOTA-conjugated commercially available peptides used for endoradiotherapy, which can readily be labeled with 177Lu or 90Y, can also accommodate 225Ac. The benefits are lower doses in normal tissue for the patient, dose reduction of the employees and environment and less shielding material. The low availability of 225Ac activity is preventing its application in clinical practice. Overcoming this barrier would open a broad field of 225Ac therapy. Independent which production pathway of 225Ac proves the most feasible, the use of automated synthesis and feasible and reproducible patient doses are needed. The Modular-Lab EAZY is one example of a GMP-compliant system, and the cassettes used for synthesis are small. Therefore, also the waste after the synthesis can be minimized. In this work, two different automated setups with different purification systems are presented. In its final configuration, three masterbatches were performed on the ML EAZY for DOTA-TATE and PSMA-I&T, respectively, fulfilling all quality criteria with final radiochemical yields of 80-90% for the 225Ac-labeled peptides.

[Development of a system for risk analysis in treatment with unsealed radioactive substances in nuclear medicine]. / Entwicklung eines Systems zur Risikoanalyse bei der Behandlung mit offenen radioaktiven Stoffen in der Nuklearmedizin.

PURPOSE: With respect to the amendment of the Radiation Protection law it is required to estimate the risks of treatments with ionizing radiation regarding patient safety. This task is in the responsibility of a medical physics expert. MATERIAL AND METHODS: The risks were estimated using the Failure Mode and Effects Analysis (FMEA) as suggested by the Federal Office for Radiation Protection (BfS) and the scientific societies. For Radioiodine Therapy, Radiosynoviorthesis, Peptide Receptor Radionuclide Therapy, and Selective Internal Radiation Therapy, respectively, the involved processes were analyzed, structured and scored. This was done both individually by all people involved in the process and by all participants together. For processes with risk priority numbers > 125 countermeasures were introduced. The risk priority number (RPZ) was calculated as the product of severity of the event, the probability of the occurrence of the event and the detection probability. RESULTS: The greatest risks were mistaken identity, incorrect estimations of activity and organ masses and a lack of compliance of the patients. The individual risk estimation revealed a high variability between the different professions, that was reduced significantly by discussion led by an external moderator. For the highest RPZ countermeasures were formulated which impact needs to be reviewed in the future. CONCLUSIONS: Nuclear medical therapies were assessed as very safe, the revealed risks are very low in respect to patient safety. FMEA method was a useful tool to identify processes with potential for optimization. The chosen procedure can be easily adopted in other nuclear medicine facilities considering structure specific aspects like technical and personal equipment and procedures. Then further risks might be detected or the here identified risk might be assessed differently.

Unexpected Bone Metastases in 99mTc-Pertechnetate Scan of Recurrent Goiter.

A 68-year-old woman who underwent a thyroidectomy for bilateral goiter 20 years ago was referred to our department. The examination revealed a newly occurred thyroid nodule in the right central lobe. Unexpectedly the Tc-pertechnetate scan revealed several extrathyroidal foci right-sided supraclavicular, beside a cold thyroid nodule in the right lower lobe. Consecutive surgery and histology confirmed the suspected diagnosis of follicular thyroid cancer with multiple bone metastases. Whole-body scintigraphy performed after the following radioiodine therapy indicated disseminated osseous metastases.

Brain Metastases of a Neuroendocrine Tumor Visualized by 68Ga-DOTATATE PET/CT.

A 65-year-old woman with a long history of metastasized, highly differentiated (G1) neuroendocrine tumor of pancreatic origin presented for follow-up Ga-DOTATATE PET/CT after 7 peptide receptor radiotherapies. In the previous scan, she already had discrete intracranial tracer accumulations, which were massively progressive in the recent Ga-DOTATATE PET/CT 8 months later. This case illustrates that cerebral metastases in neuroendocrine tumors may occur many years after initial diagnosis, and their somatostatin receptor expression may rise within a few months to a level that reasonably justifies further peptide receptor radiotherapy.