Diagnostic Accuracy of Salivary Metanephrines in Pheochromocytomas and Paragangliomas.

BACKGROUND: Measurements of plasma free metanephrines are recommended for diagnosing pheochromocytomas and paragangliomas (PPGL). Metanephrines can be detected in saliva with LC-MS/MS with sufficient analytical sensitivity and precision. Because collecting saliva is noninvasive and less cumbersome than plasma or urine sampling, we assessed the diagnostic accuracy of salivary metanephrines in diagnosing PPGL. METHODS: This 2-center study included 118 healthy participants (44 men; mean age: 33 years (range: 19--74 years)), 44 patients with PPGL, and 54 patients suspected of PPGL. Metanephrines were quantified in plasma and saliva using LC-MS/MS. Diagnostic accuracy; correlation between plasma and salivary metanephrines; and potential factors influencing salivary metanephrines, including age, sex, and posture during sampling, were assessed. RESULTS: Salivary metanephrines were significantly higher in patients with PPGL compared with healthy participants (metanephrine (MN): 0.19 vs 0.09 nmol/L, P < 0.001; normetanephrine (NMN): 2.90 vs 0.49 nmol/L, P < 0.001). The diagnostic sensitivity and specificity of salivary metanephrines were 89% and 87%, respectively. Diagnostic accuracy of salivary metanephrines was 88%, with an area under the ROC curve of 0.880. We found a significant correlation between plasma and salivary metanephrines (Pearson correlation coefficient: MN, 0.86, P < 0.001; NMN, 0.83, P < 0.001). Salivary NMN concentrations were higher when collected in a seated position compared with supine (P < 0.001) and increased with age (P < 0.001). CONCLUSIONS: Salivary metanephrines are a promising tool in the biochemical diagnosis of PPGL. Salivary metanephrines correlate with plasma free metanephrines and are increased in patients with PPGL. At this time, however, salivary metanephrines cannot replace measurement of plasma free metanephrines.

Efficacy of α-Blockers on Hemodynamic Control during Pheochromocytoma Resection: A Randomized Controlled Trial.

CONTEXT: Pretreatment with α-adrenergic receptor blockers is recommended to prevent hemodynamic instability during resection of a pheochromocytoma or sympathetic paraganglioma (PPGL). OBJECTIVE: To determine which type of α-adrenergic receptor blocker provides the best efficacy. DESIGN: Randomized controlled open-label trial (PRESCRIPT; ClinicalTrials.gov NCT01379898). SETTING: Multicenter study including 9 centers in The Netherlands. PATIENTS: 134 patients with nonmetastatic PPGL. INTERVENTION: Phenoxybenzamine or doxazosin starting 2 to 3 weeks before surgery using a blood pressure targeted titration schedule. Intraoperative hemodynamic management was standardized. MAIN OUTCOME MEASURES: Primary efficacy endpoint was the cumulative intraoperative time outside the blood pressure target range (ie, SBP >160 mmHg or MAP <60 mmHg) expressed as a percentage of total surgical procedure time. Secondary efficacy endpoint was the value on a hemodynamic instability score. RESULTS: Median cumulative time outside blood pressure targets was 11.1% (interquartile range [IQR]: 4.3-20.6] in the phenoxybenzamine group compared to 12.2% (5.3-20.2)] in the doxazosin group (P = .75, r = 0.03). The hemodynamic instability score was 38.0 (28.8-58.0) and 50.0 (35.3-63.8) in the phenoxybenzamine and doxazosin group, respectively (P = .02, r = 0.20). The 30-day cardiovascular complication rate was 8.8% and 6.9% in the phenoxybenzamine and doxazosin group, respectively (P = .68). There was no mortality after 30 days. CONCLUSIONS: The duration of blood pressure outside the target range during resection of a PPGL was not different after preoperative treatment with either phenoxybenzamine or doxazosin. Phenoxybenzamine was more effective in preventing intraoperative hemodynamic instability, but it could not be established whether this was associated with a better clinical outcome.

Clinical implications of the oncometabolite succinate in SDHx-mutation carriers.

Succinate dehydrogenase (SDH) mutations lead to the accumulation of succinate, which acts as an oncometabolite. Germline SDHx mutations predispose to paraganglioma (PGL) and pheochromocytoma (PCC), as well as to renal cell carcinoma and gastro-intestinal stromal tumors. The SDHx genes were the first tumor suppressor genes discovered which encode for a mitochondrial enzyme, thereby supporting Otto Warburg's hypothesis in 1926 that a direct link existed between mitochondrial dysfunction and cancer. Accumulation of succinate is the hallmark of tumorigenesis in PGL and PCC. Succinate accumulation inhibits several α-ketoglutarate dioxygenases, thereby inducing the pseudohypoxia pathway and causing epigenetic changes. Moreover, SDH loss as a consequence of SDHx mutations can lead to reprogramming of cell metabolism. Metabolomics can be used as a diagnostic tool, as succinate and other metabolites can be measured in tumor tissue, plasma and urine with different techniques. Furthermore, these pathophysiological characteristics provide insight into therapeutic targets for metastatic disease. This review provides an overview of the pathophysiology and clinical implications of oncometabolite succinate in SDHx mutations.

Emerging role of dopamine in neovascularization of pheochromocytoma and paraganglioma.

Dopamine is a catecholamine that acts both as a neurotransmitter and as a hormone, exerting its functions via dopamine (DA) receptors that are present in a broad variety of organs and cells throughout the body. In the circulation, DA is primarily stored in and transported by blood platelets. Recently, the important contribution of DA in the regulation of angiogenesis has been recognized. In vitro and in vivo studies have shown that DA inhibits angiogenesis through activation of the DA receptor type 2. Overproduction of catecholamines is the biochemical hallmark of pheochromocytoma (PCC) and paraganglioma (PGL). The increased production of DA has been shown to be an independent predictor of malignancy in these tumors. The precise relationship underlying the association between DA production and PCC and PGL behavior needs further clarification. Herein, we review the biochemical and physiologic aspects of DA with a focus on its relations with VEGF and hypoxia inducible factor related angiogenesis pathways, with special emphasis on DA producing PCC and PGL.-Osinga, T. E., Links, T. P., Dullaart, R. P. F., Pacak, K., van der Horst-Schrivers, A. N. A., Kerstens, M. N., Kema, I. P. Emerging role of dopamine in neovascularization of pheochromocytoma and paraganglioma.

Calculating the optimal surveillance for head and neck paraganglioma in SDHB-mutation carriers.

Germline mutations of the gene encoding succinate dehydrogenase subunit B (SDHB) predispose to head-and-neck-paraganglioma (HNPGL), sympathetic PGL, pheochromocytoma and renal cell carcinoma for which regular surveillance is required. SDHB-associated tumors harbor germline and somatic mutations, consistent with Knudson's two-hit hypothesis. To assess the penetrance and optimal surveillance for different manifestations of SDHB mutation carriers. This study included all SDHB mutation carriers who were followed at the Department of Endocrinology at the University Medical Center of Groningen. Kaplan-Meier curves were used to assess the penetrance. Poisson process was used to assess the optimal age to start surveillance and intervals. Ninety-one SDHB-mutation carriers (38 men and 53 women) were included. Twenty-seven mutation carriers (30 %) had manifestations, with an overall penetrance 35 % at the age of 60 years. We calculated that optimal surveillance for HNPGL could start from an age of 27 years with an interval of 3.2 years. This study underscores the relatively low penetrance of disease in SDHB mutation carriers. Use of the Poisson approach provides a more accurate estimation of the age to initiate surveillance and length of intervals for HNPGL. These results may give rise to reconsider the current guidelines regarding the screening of these mutation carriers.

No influence of antihypertensive agents on plasma free metanephrines.

BACKGROUND: Hypertension can be the predominant sign of pheochromocytoma (PCC) and sympathetic paraganglioma (sPGL) and screening for PCC/sPGL is often performed in patients who are already being treated with antihypertensive agents. There is very little information about the influence of antihypertensive drugs on plasma free metanephrines. The aim of this study was to determine whether commonly prescribed antihypertensive drugs can falsely elevate plasma free metanephrines concentrations measured by LC-MS/MS analysis. METHODS: In a prospective study we included patients with newly diagnosed hypertension, who started monotherapy with an antihypertensive agent (i.e. ß-blocker, thiazide diuretic or angiotensin-converting enzyme (ACE) inhibitor). Plasma free metanephrine (MN) and normetanephrine (NMN) levels were measured before and one month after the start of the medication quantified by LC-MS/MS. RESULTS: Between 2009 and 2014, 39 patients were included (ß-blocker n=13, thiazide diuretic n=14 and ACE inhibitor n=12). In the whole group, the median plasma free MN and NMN concentrations at baseline were 0.19 [0.17-0.26] nmol/L and 0.56 [0.38-0.95] nmol/L. One month after the start of antihypertensive treatment, the median plasma free MN and NMN concentrations were comparable; 0.20 [0-16-0.24] nmol/L and 0.63 [0.39-0.75] nmol/L, respectively (P=0.43 and P=0.39). Separate analysis for each of the three antihypertensive agents examined did not reveal any significant changes in the median plasma free MN and NMN concentrations. CONCLUSIONS: The measurement of plasma free MN and NMN with LC-MS/MS is not affected by use of ß-blockers, diuretics and ACE inhibitors. Withdrawal of these drugs prior to the quantification of plasma metanephrines is therefore not necessary.

Mass spectrometric quantification of salivary metanephrines-A study in healthy subjects.

BACKGROUND: Determination of metanephrine (MN), normetanephrine (NMN), and 3-methoxytyramine (3-MT) in saliva may offer potential diagnostic advantages in diagnosing pheochromocytoma. METHODS: In this preliminary study, we determined metanephrine concentrations in saliva of healthy subjects and the relationship with simultaneously measured plasma metanephrines. We also studied the possible influence of pre-analytical conditions such as a collection device, awakening, posture, and eating on the salivary metanephrine levels. RESULTS: Eleven healthy subjects were included. Fasting blood and saliva samples were collected in seated position and after 30min of horizontal rest. Plasma and salivary MN, NMN, and 3-MT concentrations were determined using a high-performance liquid chromatography tandem mass spectrometric technique (LC-MS/MS) with automated solid phase extraction sample preparation. Metanephrines were detectable in saliva from all participants both in seated and supine position. No significant correlations were observed between the MN, NMN, and 3-MT concentrations in saliva and plasma in seated or supine position. Furthermore, there was no difference between MN, NMN, and 3-MT samples collected with or without a collection device. CONCLUSION: Metanephrines can be detected in saliva with LC-MS/MS with sufficient sensitivity and precision. Our findings warrant evaluation of salivary metanephrine measurement as a novel laboratory tool in the work-up of patients suspected of having a pheochromocytoma.

Dopamine concentration in blood platelets is elevated in patients with head and neck paragangliomas.

BACKGROUND: Plasma 3-methoxytyramine (3-MT), a metabolite of dopamine, is elevated in up to 28% of patients with head and neck paragangliomas (HNPGLs). As free dopamine is incorporated in circulating platelets, we determined dopamine concentration in platelets in patients with a HNPGL. METHODS: A single center cohort study was performed between 2012 and 2014. Thirty-six patients with a HNPGL were compared to healthy controls (68 for dopamine in platelets and 120 for plasma 3-MT). RESULTS: Dopamine concentration in platelets was elevated in HNPGL patients compared to healthy controls (median [interquartile ranges] 0.48 [0.32-0.82] pmol/109 platelets vs. 0.31 [0.24-0.47] pmol/109 platelets; p<0.05), whereas plasma 3-MT concentration did not differ between both groups (0.06 [0.06-0.08] nmol/L vs. 0.06 [0.06-0.06] nmol/L; p=0.119). Based on 68 healthy controls, the reference interval for dopamine concentration in platelets was 0.12-0.97 pmol/109 platelets. Six (16.7%) patients with a HNPGL demonstrated an increased dopamine concentration in platelets compared to three (8.3%) patients with an increased plasma 3-MT level (p=0.053). The sensitivity and specificity were 16.7% and 98.5% for platelet dopamine and 8.3% and 97.5% for plasma 3-MT concentration (p=0.37). CONCLUSIONS: Dopamine concentration in platelets is elevated in patients with a HNPGL compared to healthy subjects, and may be a novel biomarker for dopamine producing paraganglioma.

Catecholamine-Synthesizing Enzymes Are Expressed in Parasympathetic Head and Neck Paraganglioma Tissue.

BACKGROUND/AIM: Increased dopamine production may be a feature of head and neck paraganglioma (HNPGL). 18F-fluorodihydroxyphenylalanine positron emission tomography scintigraphy has a high sensitivity for detecting HNPGLs. These observations strongly suggest that HNPGLs have the capacity for L-3,4-dihydroxyphenylalanine uptake and conversion towards dopamine. Therefore, our aim was to demonstrate the presence of catecholamine-synthesizing enzymes, i.e. tyrosine hydroxylase (TH), aromatic L-amino acid decarboxylase (AADC) and dopamine ß-hydroxylase (DBH) in HNPGL tissue. METHODS: A single-center study was performed among patients who underwent surgery for HNPGL at a single university referral center between 1994 and 2012. HNPGL tissue was immunohistochemically stained for TH, AADC and DBH. Data on paraganglioma-associated germline mutations, preoperative biochemical phenotype and imaging studies were retrieved. Catecholamine excess was defined as preoperative plasma and/or urinary levels of metanephrine, normetanephrine or 3-methoxytyramine above the upper reference limit. RESULTS: Nineteen HNPGLs from 18 patients were evaluated. All tumor tissues (100%) stained positive for AADC, 6 (32%) for TH and 2 (11%) for DBH. Of 3 HNPGLs staining positive for DBH, 2 were also positive for AADC and TH. Catecholamine excess was only present in 1 patient (5%). The HNPGLs of this single patient only showed positive staining for AADC. CONCLUSIONS: Catecholamine-synthesizing enzymes, in particular AADC, are expressed in the majority of HNPGL tissues.

Unilateral and bilateral adrenalectomy for pheochromocytoma requires adjustment of urinary and plasma metanephrine reference ranges.

CONTEXT: Follow-up after adrenalectomy for pheochromocytoma is recommended because of a recurrence risk. During follow-up, plasma and/or urinary metanephrine (MN) and normetanephrine (NMN) are interpreted using reference ranges obtained in healthy subjects. OBJECTIVE: Because adrenalectomy may decrease epinephrine production, we compared MN and NMN concentrations in patients after adrenalectomy to concentrations in a healthy reference population. DESIGN: A single-center cohort study was performed in pheochromocytoma patients after adrenalectomy between 1980 and 2011. SUBJECTS: Seventy patients after unilateral and 24 after bilateral adrenalectomy were included. MAIN OUTCOME MEASURES: Plasma-free and urinary-deconjugated MN and NMN determined at 3 to 6 months and annually until 5 years after adrenalectomy were compared with concentrations in a reference population. Data are presented in median (interquartile range). RESULTS: Urinary and plasma MN concentrations 3 to 6 months after unilateral adrenalectomy were lower compared with the reference population (39 [31-53] µmol/mol creatinine and 0.14 [0.09-0.18] nmol/L vs 61 [49-74] µmol/mol creatinine and 0.18 [0.13-0.23] nmol/L, respectively, both P < .05). Urinary MN after bilateral adrenalectomy was reduced even further (7 [1-22] µmol/mol creatinine; P < .05). Urinary and plasma NMN were higher after unilateral adrenalectomy (151 [117-189] µmol/mol creatinine and 0.78 [0.59-1.00] nmol/L vs 114 [98-176] µmol/mol creatinine and 0.53 [0.41-0.70] nmol/L; both P < .05). Urinary NMN after bilateral adrenalectomy was higher (177 [106-238] µmol/mol creatinine; P < .05). Changes in urinary and plasma MNs persisted during follow-up. CONCLUSION: Concentrations of MN are decreased, whereas NMN concentrations are increased after unilateral and bilateral adrenalectomy. Adjusted reference values for MN and NMN are needed in the postsurgical follow-up of pheochromocytoma patients.

Dopamine excess in patients with head and neck paragangliomas.

AIM: This study aimed to determine the prevalence of excess dopamine in relation to clinical symptoms and nuclear imaging in head and neck paraganglioma (PGL) patients. PATIENTS AND METHODS: Thirty-six consecutive patients with head and neck PGLs, evaluated between 1993 and 2009, were included. Clinical symptoms, dopamine excess (urinary 3-methoxytyramine (3-MT) or dopamine and/or plasma dopamine or 3-MT) and (nor)epinephrine excess (urinary (nor)metanephrine) as well as (111)In-octreotide and (123)I-metaiodobenzylguanide (MIBG) scintigraphy were documented. RESULTS: Dopamine excess was found in seven patients (19.4%), but was unrelated to clinical signs and symptoms. Excretion of other catecholamines was unremarkable, except in one patient with adrenal pheochromocytoma. (123)I-MIBG uptake (present in 36.1% of patients) was associated with dopamine excess (p = 0.03). CONCLUSION: Dopamine excess is present in a considerable percentage of patients with head and neck PGL, and its measurement may be useful in follow-up. Measurement of other catecholamines is necessary to rule out co-existent pheochromocytoma.