The most important decision in the diagnosis of Cushing syndrome is deciding which patients need to begin a diagnostic evaluation. With the growing epidemic of obesity and metabolic syndrome (central obesity with hypertension and insulin resistance), many patients have a Cushingoid phenotype, but most do not have Cushing syndrome.
Important considerations include female sex, as women have a higher proportion of Cushing syndrome, unexplained hypertension (particularly in young patients), new onset of glucose intolerance or diabetes, unexplained weight gain, unexplained fractures (due to premature osteoporosis), hirsutism, menstrual irregularities, and unexplained proximal muscle weakness. Cushing syndrome creates a hypercoagulable state and is associated with an increased risk of venous thromboembolic disease, such as deep vein thrombosis and pulmonary embolism.[1]Lacroix A, Feelders RA, Stratakis CA, et al. Cushing's syndrome. Lancet. 2015 Aug 29;386(9996):913-27.
http://www.ncbi.nlm.nih.gov/pubmed/26004339?tool=bestpractice.com
History
All patients with suspected Cushing syndrome should have a complete history to exclude the use of oral, injectable, inhaled, or topical glucocorticoids manifesting as iatrogenic disease. At a minimum, patients with the following conditions should go on to be screened:[23]Nieman LK, Biller BM, Findling JW, et al. The diagnosis of Cushing's syndrome: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. 2008 May;93(5):1526-40.
https://academic.oup.com/jcem/article/93/5/1526/2598096
http://www.ncbi.nlm.nih.gov/pubmed/18334580?tool=bestpractice.com
[24]Brzana J, Yedinak CG, Hameed N, et al. Polycystic ovarian syndrome and Cushing's syndrome: a persistent diagnostic quandary. Eur J Obstet Gynecol Reprod Biol. 2014 Apr;175:145-8.
http://www.ncbi.nlm.nih.gov/pubmed/24491275?tool=bestpractice.com
Features unusual for age (i.e., osteoporosis or hypertension in young patients)
Less-usual features, such as unexplained psychiatric symptoms (including depression)
Unexplained nephrolithiasis
Multiple or progressive symptoms
Polycystic ovary syndrome
Pituitary adenomas
Adrenal adenomas.[25]Zeiger MA, Thompson GB, Duh QY, et al. American Association of Clinical Endocrinologists and American Association of Endocrine Surgeons medical guidelines for the management of adrenal incidentalomas: executive summary of recommendations. Endocr Pract. 2009 Jul-Aug;15(5):450-3.
http://www.ncbi.nlm.nih.gov/pubmed/19632968?tool=bestpractice.com
Physical
Clinical features suggesting Cushing syndrome include:
Progressive proximal muscle weakness
Bruising without obvious trauma
Facial plethora or rounding
Violaceous striae
Supraclavicular fat pad
Dorsocervical fat pad.
Children exhibiting weight gain with linear decreased growth velocity should also be considered. In addition, many patients have increased frequency of acne on the face, back, and chest.
Rapid virilisation in females (rapid-onset or increased hirsutism, voice deepening, and clitoral enlargement) in the setting of cortisol excess suggest adrenal carcinoma, which is associated with a 50% to 60% chance of Cushing syndrome.[10]Fassnacht M, Dekkers OM, Else T, et al. European Society of Endocrinology clinical practice guidelines on the management of adrenocortical carcinoma in adults, in collaboration with the European Network for the Study of Adrenal Tumors. Eur J Endocrinol. 2018 Oct 1;179(4):G1-46.
https://eje.bioscientifica.com/view/journals/eje/179/4/EJE-18-0608.xml
http://www.ncbi.nlm.nih.gov/pubmed/30299884?tool=bestpractice.com
Initial biochemical testing
One of the following three high-sensitivity tests, or a combination, should be used as a first-line diagnostic test in patients with suspected Cushing syndrome:[23]Nieman LK, Biller BM, Findling JW, et al. The diagnosis of Cushing's syndrome: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. 2008 May;93(5):1526-40.
https://academic.oup.com/jcem/article/93/5/1526/2598096
http://www.ncbi.nlm.nih.gov/pubmed/18334580?tool=bestpractice.com
[26]Fleseriu M, Auchus R, Bancos I, et al. Consensus on diagnosis and management of Cushing's disease: a guideline update. Lancet Diabetes Endocrinol. 2021 Dec;9(12):847-75.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8743006
http://www.ncbi.nlm.nih.gov/pubmed/34687601?tool=bestpractice.com
[27]Nieman L. Cushing's: update on signs, symptoms and biochemical screening. Eur J Endocrinol. 2015 Oct;173(4):M33-8.
https://eje.bioscientifica.com/view/journals/eje/173/4/M33.xml
http://www.ncbi.nlm.nih.gov/pubmed/26156970?tool=bestpractice.com
Late-night salivary cortisol
Overnight 1 mg dexamethasone suppression testing
24-hour urinary free cortisol
The 48-hour 2 mg dexamethasone suppression test is rarely used in isolation but can be used in combination with other tests. Although all of the included diagnostic tests are highly sensitive and specific, dexamethasone suppression testing was found to be the most sensitive while 24-hour urinary free cortisol was less sensitive.[28]Galm BP, Qiao N, Klibanski A, et al. Accuracy of laboratory tests for the diagnosis of Cushing syndrome. J Clin Endocrinol Metab. 2020 Jun 1;105(6):2081-94.
https://www.doi.org/10.1210/clinem/dgaa105
http://www.ncbi.nlm.nih.gov/pubmed/32133504?tool=bestpractice.com
Confirmation of biochemical testing
To increase diagnostic accuracy, the initial high-sensitivity test should be repeated.[23]Nieman LK, Biller BM, Findling JW, et al. The diagnosis of Cushing's syndrome: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. 2008 May;93(5):1526-40.
https://academic.oup.com/jcem/article/93/5/1526/2598096
http://www.ncbi.nlm.nih.gov/pubmed/18334580?tool=bestpractice.com
For example, late-night salivary cortisol samples should be obtained on two separate nights.[29]Carroll TB, Raff H, Findling JW. Late-night salivary cortisol for the diagnosis of Cushing syndrome: a meta-analysis. Endocr Pract. 2009 May-Jun;15(4):335-42.
http://www.ncbi.nlm.nih.gov/pubmed/19502211?tool=bestpractice.com
[30]Raff H. Utility of salivary cortisol measurements in Cushing's syndrome and adrenal insufficiency. J Clin Endocrinol Metab. 2009 Oct;94(10):3647-55.
https://academic.oup.com/jcem/article/94/10/3647/2596462
http://www.ncbi.nlm.nih.gov/pubmed/19602555?tool=bestpractice.com
[31]Zhang Q, Dou J, Gu W, et al. Reassessing the reliability of the salivary cortisol assay for the diagnosis of Cushing syndrome. J Int Med Res. 2013 Oct;41(5):1387-94.
https://journals.sagepub.com/doi/full/10.1177/0300060513498017
http://www.ncbi.nlm.nih.gov/pubmed/24065452?tool=bestpractice.com
Similarly, at least two 24-hour urinary free cortisol samples should be collected.[32]Petersenn S, Newell-Price J, Findling JW, et al. High variability in baseline urinary free cortisol values in patients with Cushing's disease. Clin Endocrinol (Oxf). 2014 Feb;80(2):261-9.
https://onlinelibrary.wiley.com/doi/full/10.1111/cen.12259
http://www.ncbi.nlm.nih.gov/pubmed/23746264?tool=bestpractice.com
Alternatively, a second high-sensitivity test may be performed: for example, supplementing a late-night salivary cortisol test with a 24-hour urinary free cortisol measurement.[33]Elias PC, Martinez EZ, Barone BF, et al. Late-night salivary cortisol has a better performance than urinary free cortisol in the diagnosis of Cushing's syndrome. J Clin Endocrinol Metab. 2014 Jun;99(6):2045-51.
https://academic.oup.com/jcem/article/99/6/2045/2537654
http://www.ncbi.nlm.nih.gov/pubmed/24628557?tool=bestpractice.com
Testing of late-night salivary cortisol is more accurate at initial diagnosis, but late-night salivary cortisol can frequently be normal in patients with recurrent/persistent disease after pituitary surgery, requiring more samples for testing.[34]Sandouk Z, Johnston P, Bunch D, et al. Variability of late-night salivary cortisol in Cushing disease: a prospective study. J Clin Endocrinol Metab. 2018 Mar 1;103(3):983-90.
https://academic.oup.com/jcem/article/103/3/983/4797116
http://www.ncbi.nlm.nih.gov/pubmed/29329418?tool=bestpractice.com
Patients with initial normal biochemical testing are unlikely to have Cushing syndrome. If signs or symptoms progress, or intermittent Cushing syndrome is suspected, repeat biochemical testing can be performed after 6 months or at a time when cortisol hypersecretion is assumed.[23]Nieman LK, Biller BM, Findling JW, et al. The diagnosis of Cushing's syndrome: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. 2008 May;93(5):1526-40.
https://academic.oup.com/jcem/article/93/5/1526/2598096
http://www.ncbi.nlm.nih.gov/pubmed/18334580?tool=bestpractice.com
Patients with any abnormal initial biochemical testing require further investigation and referral to an endocrinologist should be considered. Before proceeding with any further evaluation, physiological causes of hypercortisolism should be excluded. These conditions include: psychiatric disorders including depression, alcohol use disorder, physical stress, malnutrition, pregnancy, and perhaps class III obesity (BMI 40 or above) or metabolic syndrome.[26]Fleseriu M, Auchus R, Bancos I, et al. Consensus on diagnosis and management of Cushing's disease: a guideline update. Lancet Diabetes Endocrinol. 2021 Dec;9(12):847-75.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8743006
http://www.ncbi.nlm.nih.gov/pubmed/34687601?tool=bestpractice.com
[35]Keller J, Flores B, Gomez RG, et al. Cortisol circadian rhythm alterations in psychotic major depression. Biol Psychiatry. 2006 Aug 1;60(3):275-81.
http://www.ncbi.nlm.nih.gov/pubmed/16458262?tool=bestpractice.com
[36]Carroll BJ, Cassidy F, Naftolowitz D, et al. Pathophysiology of hypercortisolism in depression. Acta Psychiatr Scand Suppl. 2007;(433):90-103.
http://www.ncbi.nlm.nih.gov/pubmed/17280575?tool=bestpractice.com
[37]Raff H, Raff JL, Findling JW. Late-night salivary cortisol as a screening test for Cushing's syndrome. J Clin Endocrinol Metab. 1998 Aug;83(8):2681-6.
https://academic.oup.com/jcem/article/83/8/2681/2660457
http://www.ncbi.nlm.nih.gov/pubmed/9709931?tool=bestpractice.com
Urine pregnancy testing should be considered to exclude pregnancy, and glucose testing may reveal concomitant glucose intolerance or diabetes.
If multiple additional tests are abnormal, the patient has Cushing syndrome and differential diagnostic testing should be undertaken. Greatly elevated dehydroepiandrosterone sulfate levels are suggestive of adrenocortical carcinoma, but are neither sensitive nor specific.
Algorithm for diagnosis of Cushing syndrome
Opens in new window[26]Fleseriu M, Auchus R, Bancos I, et al. Consensus on diagnosis and management of Cushing's disease: a guideline update. Lancet Diabetes Endocrinol. 2021 Dec;9(12):847-75.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8743006
http://www.ncbi.nlm.nih.gov/pubmed/34687601?tool=bestpractice.com
Initial differential diagnostic testing
Once hypercortisolism has been established, further testing to determine the aetiology should be sought.
Morning plasma adrenocorticotrophic hormone (ACTH) is the test of choice for differentiating ACTH-dependent from ACTH-independent Cushing syndrome. Suppressed ACTH levels (<1 picomol/L [<5 picograms/mL]) suggest ACTH-independent Cushing syndrome (although assays differ between different laboratories), and further investigations should include imaging of the adrenal glands to identify adrenal pathology causing hypercortisolism, such as an adenoma.
Unsuppressed ACTH levels (>4 picomol/L [>20 picograms/mL]) suggest ACTH-dependent Cushing syndrome (although assays differ between different laboratories).[1]Lacroix A, Feelders RA, Stratakis CA, et al. Cushing's syndrome. Lancet. 2015 Aug 29;386(9996):913-27.
http://www.ncbi.nlm.nih.gov/pubmed/26004339?tool=bestpractice.com
Further investigations in such patients should include pituitary/sellar magnetic resonance imaging (MRI) to identify an ACTH-secreting pituitary adenoma. A spoiled-gradient echo 3D T1 sequence has been shown to have higher sensitivity than dynamic MRI for detecting and localising pituitary micro-adenomas in patients with Cushing syndrome.[38]Grober Y, Grober H, Wintermark M, et al. Comparison of MRI techniques for detecting microadenomas in Cushing's disease. J Neurosurg. 2017 Apr 28:1-7.
http://www.ncbi.nlm.nih.gov/pubmed/28452619?tool=bestpractice.com
Patients with ACTH-dependent Cushing syndrome and an adenoma ≥10 mm on MRI should proceed to treatment. Some clinicians prefer additional biochemical confirmation with high-dose dexamethasone suppression testing before initiating surgical therapy.[39]Newell-Price J, Trainer P, Besser M, et al. The diagnosis and differential diagnosis of Cushing's syndrome and pseudo-Cushing's states. Endocr Rev. 1998 Oct;19(5):647-72.
https://academic.oup.com/edrv/article/19/5/647/2530832
http://www.ncbi.nlm.nih.gov/pubmed/9793762?tool=bestpractice.com
The use of high-dose dexamethasone suppression testing is an area of debate because of its variable sensitivity and specificity.[40]Aron DC, Raff H, Findling JW. Effectiveness versus efficacy: the limited value in clinical practice of high dose dexamethasone suppression testing in the differential diagnosis of adrenocorticotropin-dependent Cushing's syndrome. J Clin Endocrinol Metab. 1997 Jun;82(6):1780-5.
https://academic.oup.com/jcem/article/82/6/1780/2656494
http://www.ncbi.nlm.nih.gov/pubmed/9177382?tool=bestpractice.com
Inferior petrosal sinus sampling
Patients with ACTH-dependent Cushing syndrome have either Cushing's disease (90% to 95%) or ectopic ACTH production (5% to 10%). Those without definitive lesions on MRI should undergo inferior petrosal sinus sampling (IPSS); this should be carried out in a specialised centre because of potential patient risk.[26]Fleseriu M, Auchus R, Bancos I, et al. Consensus on diagnosis and management of Cushing's disease: a guideline update. Lancet Diabetes Endocrinol. 2021 Dec;9(12):847-75.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8743006
http://www.ncbi.nlm.nih.gov/pubmed/34687601?tool=bestpractice.com
Patients with adenomas 6-9 mm on MRI should have IPSS to confirm the diagnosis.[26]Fleseriu M, Auchus R, Bancos I, et al. Consensus on diagnosis and management of Cushing's disease: a guideline update. Lancet Diabetes Endocrinol. 2021 Dec;9(12):847-75.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8743006
http://www.ncbi.nlm.nih.gov/pubmed/34687601?tool=bestpractice.com
Up to 40% of patients with Cushing's disease will not have visible lesions on pituitary/sellar MRI. Patients without definitive lesions on MRI should also undergo inferior petrosal sinus sampling.[1]Lacroix A, Feelders RA, Stratakis CA, et al. Cushing's syndrome. Lancet. 2015 Aug 29;386(9996):913-27.
http://www.ncbi.nlm.nih.gov/pubmed/26004339?tool=bestpractice.com
[41]Raff H, Findling JW. A physiologic approach to diagnosis of the Cushing syndrome. Ann Intern Med. 2003 Jun 17;138(12):980-91.
http://www.ncbi.nlm.nih.gov/pubmed/12809455?tool=bestpractice.com
IPSS is the only test with sufficient diagnostic accuracy to differentiate Cushing's disease from ectopic ACTH production.[42]Oldfield EH, Doppman JL, Nieman LK, et al. Petrosal sinus sampling with and without corticotropin-releasing hormone for the differential diagnosis of Cushing's syndrome. N Engl J Med. 1991 Sep 26;325(13):897-905.
https://www.nejm.org/doi/full/10.1056/NEJM199109263251301
http://www.ncbi.nlm.nih.gov/pubmed/1652686?tool=bestpractice.com
[43]Findling JW, Kehoe ME, Shaker JL, et al. Routine inferior petrosal sinus sampling in the differential diagnosis of adrenocorticotropin (ACTH)-dependent Cushing's syndrome: early recognition of the occult ectopic ACTH syndrome. J Clin Endocrinol Metab. 1991 Aug;73(2):408-13.
http://www.ncbi.nlm.nih.gov/pubmed/1649842?tool=bestpractice.com
Patients with an IPSS central/peripheral gradient >2:1 or 3:1 after corticotrophin-releasing hormone (CRH) stimulation have Cushing's disease and can undergo therapy. Those without a gradient are likely to have ectopic ACTH production.[41]Raff H, Findling JW. A physiologic approach to diagnosis of the Cushing syndrome. Ann Intern Med. 2003 Jun 17;138(12):980-91.
http://www.ncbi.nlm.nih.gov/pubmed/12809455?tool=bestpractice.com
Tests for ectopic ACTH syndrome
Patients without an IPSS central/peripheral gradient >2:1 or 3:1 after CRH stimulation should be investigated for ectopic ACTH secretion. This evaluation generally includes computed tomography (CT) scanning of the chest, abdomen, and pelvis to look for a tumour secreting ACTH. The most common tumours that secrete ACTH are bronchial or thymic carcinoids. Other neuroendocrine tumours include islet cell and medullary thyroid cancer.[43]Findling JW, Kehoe ME, Shaker JL, et al. Routine inferior petrosal sinus sampling in the differential diagnosis of adrenocorticotropin (ACTH)-dependent Cushing's syndrome: early recognition of the occult ectopic ACTH syndrome. J Clin Endocrinol Metab. 1991 Aug;73(2):408-13.
http://www.ncbi.nlm.nih.gov/pubmed/1649842?tool=bestpractice.com
An MRI of the chest may be helpful in selected cases; other imaging modalities such as fluorodeoxyglucose positron emission tomography, gallium-68 DOTATATE positron emission tomography/CT, and octreotide scanning may be considered in some patients.[44]Doppman JL, Pass HI, Nieman LK, et al. Detection of ACTH-producing bronchial carcinoid tumors: MR imaging vs CT. AJR Am J Roentgenol. 1991 Jan;156(1):39-43.
https://www.ajronline.org/doi/pdf/10.2214/ajr.156.1.1845787
http://www.ncbi.nlm.nih.gov/pubmed/1845787?tool=bestpractice.com
[45]Pacak K, Ilias I, Chen CC, et al. The role of (18)F-fluorodeoxyglucose positron emission tomography and (111)In-diethylenetriaminepentaacetate-D-Phe-pentetreotide scintigraphy in the localization of ectopic adrenocorticotropin-secreting tumors causing Cushing's syndrome. J Clin Endocrinol Metab. 2004 May;89(5):2214-21.
https://academic.oup.com/jcem/article/89/5/2214/2844406
http://www.ncbi.nlm.nih.gov/pubmed/15126544?tool=bestpractice.com
[46]Panagiotidis E, Alshammari A, Michopoulou S, et al. Comparison of the impact of 68Ga-DOTATATE and 18F-FDG PET/CT on clinical management in patients with neuroendocrine tumors. J Nucl Med. 2017 Jan;58(1):91-6.
http://jnm.snmjournals.org/content/58/1/91.long
http://www.ncbi.nlm.nih.gov/pubmed/27516446?tool=bestpractice.com
[47]Santhanam P, Taieb D, Giovanella L, et al. PET imaging in ectopic Cushing syndrome: a systematic review. Endocrine. 2015 Nov;50(2):297-305.
http://www.ncbi.nlm.nih.gov/pubmed/26206753?tool=bestpractice.com
[48]Barrio M, Czernin J, Fanti S, et al. The impact of somatostatin receptor-directed PET/CT on the management of patients with neuroendocrine tumor: a systematic review and meta-analysis. J Nucl Med. 2017 May;58(5):756-61.
http://www.ncbi.nlm.nih.gov/pubmed/28082438?tool=bestpractice.com
Mild autonomous Cushing syndrome
These patients have an incidentally discovered adrenal adenoma producing mildly excessive cortisol (previously known as subclinical Cushing syndrome). Mild cortisol excess requires a specialised approach and may be more difficult to diagnose as a function of the inherent nature of the disease.[49]Delivanis DA, Athimulam S, Bancos I. Modern management of mild autonomous cortisol secretion. Clin Pharmacol Ther. 2019 Dec;106(6):1209-21.
http://www.ncbi.nlm.nih.gov/pubmed/31206616?tool=bestpractice.com
In patients with incidentally discovered adrenal nodules without clinical features of Cushing syndrome, use the 1 mg dexamethasone suppression test as the initial diagnostic test because urinary free cortisol and late-night salivary cortisol have a lower sensitivity in these patients.[27]Nieman L. Cushing's: update on signs, symptoms and biochemical screening. Eur J Endocrinol. 2015 Oct;173(4):M33-8.
https://eje.bioscientifica.com/view/journals/eje/173/4/M33.xml
http://www.ncbi.nlm.nih.gov/pubmed/26156970?tool=bestpractice.com
[50]Masserini B, Morelli V, Bergamaschi S, et al. The limited role of midnight salivary cortisol levels in the diagnosis of subclinical hypercortisolism in patients with adrenal incidentaloma. Eur J Endocrinol. 2009 Jan;160(1):87-92.
https://eje.bioscientifica.com/view/journals/eje/160/1/87.xml
http://www.ncbi.nlm.nih.gov/pubmed/18835977?tool=bestpractice.com
[51]Pivonello R, De Martino MC, Colao A. How should patients with adrenal incidentalomas be followed up? Lancet Diabetes Endocrinol. 2014 May;2(5):352-4.
http://www.ncbi.nlm.nih.gov/pubmed/24795241?tool=bestpractice.com
[52]Fassnacht M, Arlt W, Bancos I, et al. Management of adrenal incidentalomas: European Society of Endocrinology clinical practice guideline in collaboration with the European Network for the Study of Adrenal Tumors. Eur J Endocrinol. 2016 Aug;175(2):G1-34.
https://eje.bioscientifica.com/doi/10.1530/EJE-16-0467
http://www.ncbi.nlm.nih.gov/pubmed/27390021?tool=bestpractice.com
One meta-analysis has shown that patients with bilateral adrenal incidentalomas present a higher prevalence of subclinical Cushing syndrome compared with patients with unilateral adrenal incidentalomas.[53]Paschou SA, Kandaraki E, Dimitropoulou F, et al. Subclinical Cushing's syndrome in patients with bilateral compared to unilateral adrenal incidentalomas: a systematic review and meta-analysis. Endocrine. 2016 Feb;51(2):225-35.
http://www.ncbi.nlm.nih.gov/pubmed/26498946?tool=bestpractice.com