In vitro comparison of renal handling and uptake of two somatostatin receptor-specific peptides labeled with indium-111

Objective  Radiolabeled receptor-specific somatostatin analogs labeled with gamma- or beta-emitting radionuclides are useful for scintigraphic imaging and/or therapy of selected neuroendocrine tumors. However, significant renal uptake may result in radiotoxicological injury of the kidney and can limit clinical application of the agents. The aim of the study was to analyze renal handling, rate, and mechanism of renal accumulation of two somatostatin receptor-targeted peptides, [DOTA⁰, Tyr³, Thr⁸]-octreotide (DOTA-TATE) and [DOTA⁰, 1-Nal³]-octreotide (DOTA-NOC), labeled with indium-111 using in vitro methods. Methods  The perfused rat kidney and freshly isolated rat renal cells were used as experimental models. The perfusion was performed in a recirculation regimen at constant pressure with solution containing bovine albumin, erythrocytes, and a mixture of essential substrates. The renal cells were isolated from rat kidneys using two-phase collagenase perfusion. Accumulation studies were used to evaluate the renal uptake of the peptides and to compare their accumulation with that of passively or actively transported model drugs. The influence of selected inhibitors of receptor-mediated endocytosis and the inhibition of energy-dependent transport processes on the uptake were also investigated using isolated renal cells. Results  The renal clearance of ¹¹¹In-DOTA-NOC in the perfused rat kidney was significantly lower than that of ¹¹¹In-DOTA-TATE. Reverse situation was found in the case of renal retention. Pretreatment of the perfused kidney with maleate markedly decreased the renal retention. ¹¹¹In-DOTA-NOC was accumulated in the isolated renal cells at a higher rate than ¹¹¹In-DOTA-TATE (ratio 3: 1). The uptake of the radiopeptides in renal cells was higher than the uptake of not only the passively transported sucrose but also actively transported and accumulated methylglucose. The rank order of potency to inhibit the uptake by active endocytosis was approximately aprotinin > maleate > lysine. The uptake of the radiopeptides in the renal cells was temperature dependent. Conclusions  Both in vitro methods showed a higher renal accumulation of ¹¹¹In-DOTA-NOC in comparison with ¹¹¹In-DOTA-TATE. The renal uptake was partly decreased by inhibitors of receptor-mediated endocytosis and by a block of energy-dependent processes. A significant participation of active transport processes in renal accumulation of the studied peptides was confirmed.     

In vivo proton MR spectroscopy of primary and nodal nasopharyngeal carcinoma

BACKGROUND AND PURPOSE: The aim of this study was to determine the feasibility of performing in vivo proton ((1)H) MR spectroscopy of nasopharyngeal carcinoma (NPC) and to document the (1)H spectrum of this cancer. METHODS: Twenty-seven patients with NPC lesions >1 cm(3) underwent localized (1)H MR spectroscopy performed at 1.5 T. Water-suppressed spectra from both primary tumors (nine cases) and metastatic nodes (18 cases) were obtained at TE 136 and 272. Spectra were analyzed in the time domain by using a nonlinear least squares fitting algorithm with incorporation of previous knowledge. Choline (Cho)/creatine (Cr) ratios for primary NPC and metastatic nodes were calculated and compared. Spectra from normal neck muscle of five volunteers were acquired as control data. RESULTS: (1)H MR spectroscopy was successfully obtained in seven (78%) of nine primary tumors and 16 (89%) of 18 metastatic nodes. Intense lipid signals in the range of 0.89 to 2.02 ppm were observed in 95% of spectra at TE 136 and 91% of spectra at TE 272. At TE 136, Cho/Cr for metastatic nodes (5.3 +/- 1.6) was significantly higher than the ratio for primary (2.6 +/- 0.5) NPC lesions (P =.02). Cho/Cr ratios for NPC lesions were higher than those for normal neck muscles, for which values ranged from 0 to 0.97 and 0 to 1.1 at TE 136 and 272, respectively. CONCLUSION: (1)H MR spectroscopy is a feasible technique for the evaluation of NPC tumors >1 cm(3). Cho/Cr ratios for the lesions were high compared with those for normal neck muscle.     

In vivo Studies of 111In-Labelled canine Lymphocytes

[¹¹¹In]oxine was used to label canine periperal blood lymphocytes from two normal animals and one animal with breast carcinoma. Using 150 μCi of [¹¹¹In]oxine to label 50 × 106 cells yielded a labeling efficiency between 40 and 75% and a viability of about 95%. Gamma camera images obtained 24–44 h after injection showed uptake in the subcutaneous lymph nodes of the head, neck, axilla, pelvis and popliteal regions. Uptake in the liver, spleen, bowel, and bone marrow was also seen. In the animal bearing breast carcinoma, uptake occurred in the tumor, lungs and in a draining sinus in the left axilla. These results suggest that ¹¹¹In labeled lymphocytes are useful in visualizing regional lymph nodes and may be valuable in assessing tumor recognition by these cells in vivo.     

Solid renal mass in the cancer patient: second primary renal cell carcinoma versus renal metastasis

Most renal metastases are asymptomatic, occur with widespread metastatic disease, and are too small to be detected with computed tomography (CT). Rarely they form large masses. These are typically angiographically hypovascular and show only minimal CT contrast enhancement. Renal carcinoma as a second primary malignancy in the cancer patient is 4.5 times more common than mass-like renal metastases and demonstrates two CT contrast enhancement patterns. The latter include either minimal enhancement or irregular regions of intense enhancement. These CT contrast enhancement patterns of both renal carcinoma and metastasis can be used to direct the further diagnostic evaluation of these masses and distinguish between a renal metastasis or a second primary renal carcinoma in the cancer patient.     

肾透明细胞癌与肾乳头状癌、嫌色细胞癌MRI表现的对照研究

目的 探讨MR动态增强扫描对肾癌亚型的鉴别诊断价值.方法 搜集77例经病理证实的肾癌患者资料,其中透明细胞癌(CCRCC)55例,乳头状癌(PRCC)14例,嫌色细胞癌(CRCC)8例,回顾性分析各亚型肿瘤患者MR平扫及动态增强扫描表现并与病理对照,根据肿瘤及肾皮质增强前后的皮质期、实质期及延迟期信号变化,分别进行百分比测量、肿瘤-肾皮质增强指数计算,并采用单因素方差分析和LSD法进行比较.结果 CRCC多数信号均匀(7/8);CCRCC及PRCC多数信号不均(分别为51/55和13/14)、常见坏死(36/55和7/14),PRCC最常见出血(9/14)及囊变(9/14).动态增强各期CCRCC强化程度最高,强化模式呈"快进快退",CRCC轻至中度强化,PRCC强化最轻,两者均呈渐进性延迟强化.CCRCC、PRCC及CRCC皮质期信号变化分别为(296.15±60.27)%、(79.70±18.84)%和(119.56±40.76)%,实质期分别为(236.33±58.31)%、(122.81±27.35)%和(163.06±33.91)%,延迟期分别为(216.83±46.72)%、(117.55±20.63)%和(179.72±32.89)%;三者皮质期的肿瘤-皮质增强指数分别为1.26±0.34、0.33±0.12及0.54±0.10,实质期分别为0.92±0.23、0.41±0.23及0.62±0.15,延迟期分别为0.76±0.14、0.35±0.11及0.69±0.12,各亚型增强各期的信号变化(F值分别为940.931、124.515、38.194,P值均＜0.01)、肿瘤-皮质增强指数(F值分别为798.625、78.308、73.699,P值均＜0.01)差异均有统计学意义.3种亚型的MRI表现与病理学所见基本相符.结论 CCRCC、PRCC及CRCC的MRI动态增强有一定特征性的表现,与其病理特点密切相关,在肾癌亚型的鉴别诊断上有着较高的临床应用价值.

Abstract：

Objective To investigate the differential diagnostic features of subtypes of renal cell carcinoma(RCC) using dynamic contrast-enhanced MRI(DCE-MRI).Methods The MRI appearances of 77 RCCs, including 55 clear cell RCCs(CCRCC),14 papillary RCCs(PRCC) and 8 chromophobe RCCs(CRCC), were retrospectively analyzed and compared with findings of pathology. DCE-MRI was conducted in each case after intravenous administration of contrast agent. Region of interest measurements (cortical, nephrographic and delayed Phases) of signals within tumor and uninvolved renal cortex were used to calculate percentage signal intensity change and tumor-to-cortex enhancement index, and the data was analyzed by AVONA and t test. Results On unenhanced and enhanced MRI, most CRCCs showed homogeneous signal(7/8). CCRCC and PRCC often show inhomogenous signal with necrosis(36/55, 7/14). Hemorrhage and cystic degeneration were often found in PRCC (9/14). On the cortical, nephrographic and delayed phase images, CCRCCs showed greater signal intensity change[(296.15±60.27)%, (236.33±58.31)% and (216.83±46.72)%,respectively than PRCCs (79.70±18.84)%, (122.81±27.35)% and (117.55±20.63)%, respectively], and CRCCs showed intermediate change [(119.56±40.76)%, (163.06±33.91)% and (179.72±32.89)%, respectively].A phenomenon of quick staining and quick fainting was observed in CCRCCs. Both of CRCCs and PRCCs showed delayed enhancement. The tumor-to-cortex enhancement index at the cortical, nephrographic and delayed phases was highest for CCRCCs (1.26±0.34, 0.92±0.23 and 0.76±0.14, respectively), lowest for PRCCs (0.33±0.12, 0.41±0.23 and 0.35±0.11, respectively), and intermediate for CRCCs (0.54±0.10, 0.62±0.15 and 0.69±0.12, respectively,P＜0.01). The degree of enhancement was significantly different among the 3 subtypes at the every contrast enhanced phase (F=940.931, 124.515 and 38.194, P＜0.01), so was the tumor-to-cortex enhancement index(F=798.625,78.308 and 73.699, P＜0.01). There was a good consistency between MR appearances of the 3 RCC subtypes and pathological characteristics. Conclusion DCE-MRI could distinctly show imaging features of CCRCC, PRCC and CRCC, which were related to their pathological characteristics, and these features were helpful in predicting a specific subtype of RCC.     

In vivo migration of 111In radiolabeled mouse-spleen lymphocytes and tumor cells

Mouse-spleen lymphocytes, mouse mammary adenocarcinoma cells (MMA-67) and mouse lymphoma cells (S49.1) were radiolabeled with 111In and their in vivo migration patterns studied over a 24-72 h period after i.v. injection into syngeneic mice. All three cell lines showed different in vivo migration patterns. Initially, some trapping of the cells occurred in the lungs at 15 min. More MMA-67 cells were initially trapped in the lungs than S49.1 cells or lymphocytes. At 24 h, most of the 111In labeled cells had left the lungs and accumulated in various tissues and organs. At 48 and 72 h, only slight changes in the localization patterns of the 111In radiolabeled cells in vivo were noted when compared to the 24 h patterns. About 35% of the injected 111In was excreted from the animals over the 72 h period of these experiments. In vitro studies compared the growth and/or viability of 111In radiolabeled lymphocytes or tumor cells to nonradiolabeled lymphocytes or tumor cells. The percentage of 111In released from the cells in culture was also determined. Similar values for growth and/or viability were obtained between non-radiolabeled cells and MMA-67 cells radiolabeled with 1.35 dpm 111In per cell; S49.1 cells, 0.51 dpm111In per cell; and lymphocytes, 0.04 dpm 111In per cell.     

In vivo SPECT/CT imaging of human orthotopic ovarian carcinoma xenografts with 111In-labeled monoclonal antibodies

Epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor 3 (VEGFR-3) are expressed in the tumor area during the progression of ovarian carcinoma. Monoclonal antibodies developed against these receptors are potential diagnostic molecules for in vivo imaging of ovarian carcinoma.

Methods

Biodistribution of the monoclonal antibodies cetuximab against EGFR and mF4-31C1 against VEGFR-3 was studied in nude mice with orthotopic SKOV-3m human ovarian carcinoma xenografts. The biodistribution of ¹¹¹Indium-labeled antibodies was followed up to 48 h postinjection using combined SPECT and CT imaging modality. Organ samples were collected postmortem and specific organ activity was measured. Accumulation of the intravenously injected antibodies in the tumor tissue and lymph nodes was verified using immunohistology.

Results

Imaging studies with SPECT/CT showed clear accumulation of both antibodies into tumor area. The tumor uptake was 8.78±0.74 %ID/g for cetuximab and 5.77±0.62 %ID/g for mF4-31C1 after 48 h postinjection. Cetuximab had lower liver tropism and faster tumor homing rate. In addition, after 48 h two of five tumor-bearing mice showed a clear accumulation of the In-labeled mF4-31C1 at the left axillary area. Both intravenously administered antibodies could also be detected from the tumor sections by immunohistological staining but only mF4-31C1 forms in the lymph nodes.

Conclusion

These results demonstrate the accumulation of EGFR- and VEGFR-3-specific antibodies in orthotopic ovarian carcinoma tumors. Systemically administered they had slow pharmacokinetics which is typical for antibodies. Accumulation of mF4-31C1 antibody in the lymph nodes suggests the remote activation of VEGFR-3 by the primary tumor.     

Multimodality approach to staging renal cell carcinoma

Renal imaging has dramatically improved since the introduction of ultrasound (US), computed tomography (CT), and most recently magnetic resonance (MR) imaging. US and MR imaging are ideal for patients with compromised renal function preventing administration of iodinated contrast material or those who have experienced reactions to contrast. Staging errors occur due to limitations in assessing microscopic tumor invasion of the renal capsule and perinephric fat, detecting metastatic deposits in normal sized lymph nodes and differentiating inflammatory hyperplastic lymph nodes from neoplastic ones. These limitations are shared by US, CT, and MR imaging. Vascular invasion by tumor can be evaluated by all imaging modalities including venography. The advantages and limitations of each examination will be presented.     

In vivo use of a radioiodinated somatostatin analogue: dynamics, metabolism, and binding to somatostatin receptor-positive tumors in man.

Somatostatin analogues, labeled with gamma-emitting radionuclides, are of potential value in the localization of somatostatin receptor-positive tumors with gamma camera imaging. We investigated the application in man of a radioiodinated analogue of somatostatin, 123I-Tyr-3-octreotide, which has similar biologic characteristics as the native peptide. The radiopharmaceutical is cleared rapidly from the circulation (up to 85% of the dose after 10 min) mainly by the liver. Liver radioactivity is rapidly excreted into the biliary system. Until 3 hr after injection, radioactivity in the circulation is mainly in the form of 123I-Tyr-3-octreotide. Thereafter, plasma samples contain increasing proportions of free iodide. Similarly, during the first hours after injection, radioactivity in the urine exists mainly in the form of the unchanged peptide. Thereafter, a progressive increase in radioiodide excretion is observed, indicating degradation of the radiopharmaceutical in vivo. Fecal excretion of radioactivity amounts to only a few percent of the dose. The calculated median effective dose equivalent is comparable with values for applications of other 123I-radiopharmaceuticals (0.019 mSv/MBq).     

Use of indium-111 pentetreotide somatostatin receptor scintigraphy to detect recurrent thyroid carcinoma in patients without detectable iodine uptake

We conducted a prospective evaluation of somatostatin receptor scintigraphy (SRS) for the diagnosis of recurrent vesicular or papillary thyroid carcinoma in 16 patients with no detectable iodine uptake. SRS was performed 1, 4 and 24 h after intravenous injection of 137–200 MBq of indium-111 pentetreotide. Results were interpreted in terms of assumed presence of tumoral tissue: there were three true-positives (19%), one false-positive (6%) and 12 false-negatives (75%). The three true-positive patients had multiple lesions visible on computerized tomography. SRS was negative in all patients with a high thyroglobulin level alone. In addition, we analyzed the consequences of interpretative criteria and somatostatin receptor expression variability for SRS positivity as well as the risk of false-positives. We conclude that when iodine uptake cannot be demonstrated in patients with suspected recurrence of differentiated thyroid carcinoma, SRS would not appear to contribute to diagnosis, and that interpretative criteria commonly used for tumours with a high receptor density may be too restrictive for tumours with a low receptor density. Received 28 January and in revised form 16 April 1998     

In vivo viability of 111In-labelled granulocytes demonstrated in a sham-dialysis model

In seven febrile patients undergoing regular dialysis treatment, sham-dialysis with a dialyzer equipped with a cuprophane membrane was performed during a routine 111In-oxine white blood cell scan with "pure" granulocytes isolated on a discontinuous gradient (Percoll/plasma: n = 5; Metrizamide/plasma: n = 2). The patients were in contact with the cuprophane membrane over 45 or 90 min. Twenty-five (+/- 5) minutes after the start of the dialysis, the peripheral leucocyte count (59 +/- 22%) and the 111In activity in the peripheral blood decreased to a minimum of their initial range (64 +/- 21%) because of leucocyte activation. The activity over both lungs increased symmetrically (29 +/- 15%), the spleen activity decreased (14.8 +/- 8%) and the liver activity remained constant. At the end of the dialysis (45-90 min post-injection), the number of circulating neutrophils, peripheral 111In activity and activity distribution in the organs regained their initial level. In conclusion, these data confirm previously described data concerning the sequestration of neutrophils into the lung during dialysis treatment. The data demonstrate the origin of the activated neutrophils in the circulation and the spleen. The identical behaviour of 111In-oxine-labelled and unlabelled granulocytes is demonstrated. Additionally, the accumulation of activity in the spleen is due not to opsonized cells but to sequestrated neutrophils, which are able to migrate from the spleen after adequate activation. The migration of activated cells into the lung explains the diagnostic difficulties posed by diffuse lung uptake in leucocyte scans in patients with leucocyte-activating diseases.     

In vivo evaluation of 111In-labeled T-lymphocyte homing in experimental colitis.

Blockade of lymphocyte recruitment to the intestinal mucosa is considered a useful therapy for inflammatory bowel disease (IBD) and anti-alpha4 antibodies have clinical benefit in patients with active Crohn's disease. The aim of this study was to evaluate a scintigraphic technique to assess lymphocyte homing to the colon in 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced experimental colitis (TNBS colitis) in vivo. METHODS: TNBS-sensitized and nonsensitized murine total lymphocytes or CD4+ lymphocytes were radiolabeled with 111In-oxinate. Cells were injected into control mice (n = 5) or mice with TNBS colitis (n = 5). Specific abdominal radioactive uptake was determined by SPECT using a dedicated pinhole system 48 h after cell transfer. Radioactive colon uptake was correlated with histology and colon weight as parameters of inflammation. RESULTS: The radioactive colon uptake was most evident in mice with TNBS colitis that received sensitized lymphocytes (uptake ratio [mean +/- SEM], 0.51 +/- 0.03 vs. 0.22 +/- 0.04; P = 0.004). The sensitized 111In-labeled lymphocytes exacerbated colitis compared with nonsensitized lymphocytes. The colon uptake correlated well with both colon weight and histologic score (R2 = 0.836 and 0.933, respectively). The use of purified 111In labeled CD4+ lymphocytes resulted in a similar scintigraphic pattern. Administration of an anti-alpha4 antibody decreased radioactivity colon uptake of the (111)In-labeled cells compared with the control antibody in mice with TNBS colitis (uptake ratio, 0.72 +/- 0.14 to 0.33 +/- 0.03; P = 0.012). CONCLUSION: Animal pinhole SPECT can be applied for temporal and spatial analysis of the lymphocyte homing process in experimental colitis. This technique makes possible the in vivo evaluation of therapeutic efficacy of new drugs that interfere with lymphocyte migration. Moreover, colon uptake of radioactivity can be used as a parameter of disease activity in experimental colitis.     

In vivo imaging of rat lymphocytes with an indium 111-labelled anti-T cell monoclonal antibody: a comparison with indium 111-labelled lymphocytes

An indium 111-labelled mouse anti-rat T cell monoclonal antibody, MRC OX-19, was injected intravenously into rats to establish the usefulness of radiolabelled anti-lymphocyte antibodies in imaging lymphoid tissues. Antibody binding in vivo, measured by immunofluorescence analysis of cell suspensions made from lymphoid tissues, was detectable on lymphocytes in blood, spleen and lymph nodes. The extent of binding was time and antibody-dose dependent. Doses of antibody above 80 g/kg body weight resulted in modulation, i.e. loss of CD 5 (T 1) molecules from the cell surface, although the cells remained in the circulation. Modulation was demonstrable within 2 h and for at least 24 h after a single injection of antibody. Intravenous injection of¹¹¹In-MRC OX-19 resulted in levels of in vivo binding comparable with those seen with unlabelled antibody. Scintillation imaging showed early splenic localisation persisting over 48h, a more gradual localisation in the lymph nodes seen clearly at 24 h and a steady background. Comparison of the in vivo distribution of labelled antibody and¹¹¹In-tropolone-labelled lymphocytes showed that both could be used for external imaging of lymphocytes by scintillation camera.     

In vivo imaging of rat lymphocytes with an indium 111-labelled anti-T cell monoclonal antibody: a comparison with indium 111-labelled lymphocytes

An indium 111-labelled mouse anti-rat T cell monoclonal antibody, MRC OX-19, was injected intravenously into rats to establish the usefulness of radiolabelled anti-lymphocyte antibodies in imaging lymphoid tissues. Antibody binding in vivo, measured by immunofluorescence analysis of cell suspensions made from lymphoid tissues, was detectable on lymphocytes in blood, spleen and lymph nodes. The extent of binding was time and antibody-dose dependent. Doses of antibody above 80 micrograms/kg body weight resulted in modulation, i.e. loss of CD 5 (T1) molecules from the cell surface, although the cells remained in the circulation. Modulation was demonstrable within 2 h and for at least 24 h after a single injection of antibody. Intravenous injection of 111In-MRC OX-19 resulted in levels of in vivo binding comparable with those seen with unlabelled antibody. Scintillation imaging showed early splenic localisation persisting over 48 h, a more gradual localisation in the lymph nodes seen clearly at 24 h and a steady background. Comparison of the in vivo distribution of labelled antibody and 111In-tropolone-labelled lymphocytes showed that both could be used for external imaging of lymphocytes by scintillation camera.     

Staging of renal cell carcinoma

As in other malignant tumors, prognosis in renal cell carcinoma (RCC) depends on tumor extent and metastasis at the time of primary diagnosis. Staging systems formalize the way in which the extent of RCC is being described and classified. Primary staging of RCC aims at evaluating surgical options. Since surgical excision, which is the mainstay of therapy in non-metastatic RCC, and, recently, minimally invasive ablation methods have evolved significantly over the last decades, staging systems continue to evolve along the way. The 40-year-old Robson classification has been replaced with the TNM classification of RCC, because the latter adapts more easily to changing patterns of diagnosis and therapy. Modern cross-sectional imaging methods, such as multidetector-row computed tomography (MDCT), and magnetic resonance imaging (MRI), perform highly in T-staging of local tumor extent and M-staging of distant metastasis. However, both MDCT and MRI perform poorly in N-staging of lymphadenopathy. At present, 18-F-desoxy-glucose positron emission tomography (FDG-PET) appears to be unreliable in the detection of RCC and its metastasis. This overview of current radiological and surgical literature attempts to describe how modern staging systems for RCC are organized, and which radiological and surgical developments currently influence the way in which primary staging and prognosis of RCC depend on one another.