Endocrinology - Adrenals Flashcards
What are the three zones of the adrenal cortex?
The adrenal cortex consists of the zona glomerulosa, the source of aldosterone (mineralocorticoid), zona fasciculata, accounting for 75% of the adrenal cortex and producing corticosteroids (glucocorticoids) and zona reticularis a narrow juxtamedullary ring of cells producing sex steroids.
What are structure are the adrenal steroids synthesised from?
All the steroids of the adrenal cortex are synthesised from cholesterol and are chemical modifications of it.
Cholesterol, progesterone, the glucocorticoids and the mineralocorticoids are all 21 carbon steroids.
Androgens are 19 carbon steroids and oestrogens (produced primarily in the ovaries) are 18 carbon steroids.
What is the basis of the specialisation of different layers of the adrenal cortex to synthesise and secrete particular steroid hormones?
Different layers of the adrenal cortex contain different enzymes responsible for the chemical modification of cholesterol. For example, the zona reticularis primarily produces androgens becayse they contain the 17,20-lyase enzyme. On the other hand, the zona glomerulosa produces aldosterone because it contains aldosterone synthase.
Where does the cholesterol needed for steroid hormone synthesis come from?
Cholesterol is the precursor to ALL steroid hormones. Most of the cholesterol is provided to the adrenal cortex via the circulation, and small amounts are synthesised de novo within the adrenal cortical cells. Cholesterol circulates bound to low density lipoproteins (LDLs). There are receptors for these lipoproteins in the membranes of adrenocortical cells. The lipoprotein-cholesterol complex binds and is enters the cell by endocytosis. Once inside the cell, cholesterol is stored in cytoplasmic vesicles until needed for steroid hormone synthesis.
What do the enzymes that catalyse cholesterol into active steroid hormones need?
They require cytochrome P450 complex, molecular oxygen and NADPH which serves as a hydrogen donor for the reducing steps. A flavoprotein enzyme called adrenodoxin reductase and an iron containing protein called adrenodoxin are intermediaries in the transfer of hydrogen from NADPH to the cyt-P450 complex enzymes.
After cholesterol what do all steroid hormones get converted to?
The first step in each pathway is the conversion of cholesterol to pregnenolone, catalysed by cholesterol desmolase. Thus, all layers of the adrenal cortex contain cholesterol desmolase. Cholesterol desmolase is the rate limiting enzyme in the pathway and is stimulated by ACTH.
What is the major glucocorticoid produced by the zona fasciculata of the adrenal cortex?
The major glucocorticoid in produced in humans is cortisol (hydrocortisone). Thus, zona fasciculata contains all the enzymes required to convert cholesterol to cortisol. These are:
1) cholesterol desmolase: converts cholesterol to pregnenolone
2) 17 alpha hydroxylase: which hydroxylates pregnenalone into 17-hydroxypregnenolone
3) 3 beta hydroxysteroid dehydrogenase: which converts 17-hydroxypregnenolone to 17-hydroxyprogesterone
4) 21 beta hydroxylase and 11 beta hydroxylase which hydroxylate C11 and C21 residues to produce the final product, cortisol
Interestingly, these do not have to happen in this exact order. For example, hydroxylation at C17 can occur before or after the action of 3 beta hydroxysteroid dehydrogenase.
Other than cortisol, what other steroid in the pathway has glucocorticoid activity? Why is this important?
Corticosterone also has glucocorticoid activity. If the 17 alpha hydroxylase step is blocked, the zona fasciculata can still produce corticosterone without any deleterious effects. Thus, cortisol is NOT absolutely necessary for life provided corticosterone is being produced.
However, blocks at the cholesterol desmolase, 3 beta hydroxysteroid dehydrogenase, 21 beta hydroxylase or 11 beta hydroxylase steps are devastating because they prevent the production of cortisol and corticosterone. In these cases, death will ensue without appropriate hormone replacement therapy.
What drugs significantly interfere with glucocorticoid biosynthesis?
Metyrapone and ketoconazole. Metyrapone inhibits 11 beta hydroxylase activity, the last step in cortisol synthesis. Ketoconazole inhibits several steps in the pathway including cholesterol desmolase, the first step.
What are the main androgens produced by the adrenal cortex?
DHEA and androstenedione are androgenic steroids produced by the zona reticularis. These compounds have weak androgenic activity, but in the testes are converted to testosterone, a more potent androgen. The precursors of adrenal androgens are 17 hydroxypregnenolone and 17 hydroxyprogesterone which are converted to androgens by the removal of the C20, 21 side chains.
In men, adrenal androgens are of little significance as the testes produce their own testosterone from cholesterol and do not require the adrenal precursors. In females, however, the adrenal cortex is a major source of androgenic compounds.
The zona reticularis also produces a very small amount of testosterone and 17 beta oestradiol, although the main sources for these hormones are the testes and ovaries respectively.
What is meant by the term 17-ketosteroids?
Adrenal androgens have a ketone group at C17 that distinguishes them from cortisol, aldosterone and testosterone. Thus, tha major adrenal androgens are called 17 ketosteroids which can be measured in the urine.
What is the major mineralocorticoid produced by the zona glomerulosa of the adrenal cortex?
The major mineralocorticoid in the body is aldosterone, which is synthesised ONLY by the zona glomerulosa. The steps required to convert cholesterol to corticosterone are identical to those in zona fasciculata and the addition of aldosterone synthase in the zona glomerulosa converts corticosterone to aldosterone.
Why does the zona glomerulosa not produce corticosteroids?
There are 2 reasons for this:
i) corticosterone, a glucocorticoid, is converted to aldosterone because this zone contains aldosterone synthase, and
ii) the zona glomerulosa lacks 17 alpha hydroxylase and therefore is unable to produce cortisol from progesterone
Is aldosterone the only steroid with mineralocorticoid activity?
No. 11-deoxycorticosterone (DOC) and corticosterone also have mineralocorticoid activity. Thus, if the mineralocorticoid pathway is blocked BELOW the level of DOC (e.g. absence of 11 beta hydroxylase or aldosterone synthase) mineralocorticoids will continue to be produced. However, if the pathway is blocked ABOVE the level of DOC (e.g. absence of 21 beta hydroxylase) then no mineralocorticoids will be produced.
Differentiate Cushing’s syndrome from Cushing’s disease
Cushing’s syndrome refers to hypercortisolaemia and its associated signs and symptoms from ANY cause.
Cushing’s disease refers specifically to hypercortisolaemia due to ACTH production by a pituitary adenoma.
The most common cause of Cushing’s syndrome is iatrogenic, owing to exogenous steroid use to treat a number of conditions. If one excludes iatrogenic hypercortisolaemia, the most common cause of Cushing’s syndrome is Cushing’s disease, which accounts for approximately 2/3’s of all cases.
What are the signs and symptoms of Cushing’s disease?
Atrophic, thin skin; easy bruising and purple striae on abdomen, axilla, hips, and thighs
Weight gain or central obesity
Dorsocervical (buffalo hump) and supraclavicular fat accumulation
Moon facies
Menstrual irregularities
Hirsutism
Diabetes or insulin resistance
Muscle weakness
Hypertension
Increased susceptibility to infection
Frequent fungal infections
Osteoporosis or osteopenia
Psychiatric symptom such as depression, mood changes, and even psychosis
Hypercoagulable state
List the four steps involved in diagnosing Cushing’s syndrome
Step 1: screen for and document hypercortisolaemia with a 24 hour urine cortisol or dexamethasone suppression test (DST)
Step 2: differentiate between ACTH dependent and ACTH independent causes by measuring ACTH
Step 3: distinguish pituitary Cushing’s disease from ectopic ACTH secretion with a pituitary MRI and/or inferior petrosal sinus sampling (IPSS)
Step 4: surgically resect the tumour once identified
When should one screen for Cushing’s syndrome?
The main indication is the in the presence of multiple symptoms of Cushing’s syndrome or worsening features of Cushing’s. However, the symptoms that could be associated with Cushing’s syndrome - e.g. depression, weight gain, hypertension, diabetes, osteoporosis - are common. Consider screening adults with weight gain, an abnormal fat distribution, proximal muscle weakness, large purple striae and new cognitive depression.
In young people, be suspicious if there is non traumatic fractures, cutaneous atrophy or hypertension.
How should one screen for Cushing’s syndrome?
The first line test of choice is the low dose (1 mg) overnight dexamethasone suppression test (DST).
24 hour urine free cortisol levels or late night salivary cortisol levels can also be undertaken.
How is the DST performed?
1mg of dexamethasone is given at 11pm and the serum cortisol is measured at 8am the following morning. If the patient does not have Cushing’s, the 8am cortisol is suppressed (due to negative feedback on the pituitary) to <1.8ug/dL.
How is the 24 hour urine free cortisol done?
By collecting urine over 24 hours to measure free cortisol. To ensure an accurate result, the urine collection should be confirmed with a complete and simultaneous urine creatinine excretion. Urinary cortisol should be normal in patients without Cushing’s syndrome. This test should be repeated if there is a normal first result but a high index of suspicion.
What is the late night salivary cortisol test?
Measurement of cortisol levels in salivary samples collected late at night. Some clinicians advocate this test because it is easy to perform and the salivary and plasma cortisol levels closely correlate. Normal cortisol levels should be low at night, confirming normal diurnal variation.
Cushing’s patients have abnormally high late night levels. This test is more useful for patients who have episodic hypercortisolaemia, with most clinicians favouring the DST or 24 hour urine free cortisol.
What second line tests can be used if the DST or 24 urinary free cortisol is normal?
1) 48h dexamethasone suppression test: give dexamethasone 0.5mg/6h PO for 2d. Measure cortisol at 0 and 48h. Again in Cushing’s syndrome there is a failure to suppress cortisol
2) 48h high dose dexamethasone suppression test: 2mg/6h may distinguish pituitary (suppression) from other causes (no/part suppression)
3) Midnight cortisol
Once hypercortisolaemia is confirmed what is the next step in evaluating a patient with Cushing’s syndrome?
After ruling out ingestion of exogenous steroids, the next step is to differentiate between an ACTH dependent (80%) and an ACTH independent (20%) disease. ACTH dependent diseases are associated with pituitary adenoma (80%), ectopic ACTH (20%) and CRH hypersecretion (rare). ACTH independent disease is associated with adrenal adenoma, adrenal carcinoma, nodular dysplasia and McCune-Albright syndrome (rare).
Measurement of ACTH and cortisol. If ACTH >10pg/mL, the patient most likely has an ACTH dependent cause of Cushing’s syndrome. In addition, an ACTH >10pg/mL after corticotrophin releasing hormone (CRH) administration suggests ACTH dependency (specifically due to a pituitary cause as cortisol rises with pituitary disease but not with ectopic ACTH).
If ACTH is undetectable how should the patient be investigated?
In this instance the likely cause is an adrenal tumour. A CT of the adrenal glands is the next step. If no mass is located then proceed to adrenal vein sampling or adrenal scintigraphy (radiolabelled cholesterol derivative).
Once ACTH-dependent Cushing’s syndrome has been confirmed, what is the final step in making the biochemical diagnosis?
A high-dose (8-mg) DST or CRH test to differentiate between a corticotrope adenoma and an ectopic ACTH-secreting tumor. Patients with a pituitary source of ACTH retain suppressibility of cortisol to high-dose dexamethasone, whereas patients with ectopic ACTH tumors do not (because the tumour is autonomous).
Cortisol rises with pituitary disease following CRH administration, whereas it remains the same with ectopic disease.
How is the dexamethasone test confirmed?
If tests indicate that cortisol responds to manipulation, Cushing’s disease is likely. Image the pituitary (MRI) and consider IPSS. This test takes advantage of the concentration gradient between pituitary venous drainage via the inferior petrosal sinus (IPSS) and peripheral venous values of ACTH to further determine whether an ACTH producing corticotropic adenoma is present in the pituitary. The inclusion of CRH adds greater sensitivity to the test.
Explain how IPSS is performed?
Samples of ACTH and cortisol are obtained simultaneously from the IPS (central) and from a peripheral site (e.g., inferior vena cava [IVC]). In patients with Cushing’s disease, the central/peripheral ratio (C/P = IPS/IVC ratio) of ACTH > 2. In patients with ectopic ACTH, the ratio < 2 and selective venous sampling (e.g., of the pulmonary, pancreatic, or intestinal beds) may localize the ectopic tumor.
What is pseudo Cushing’s?
A clinical state characterized by mild overactivity of the hypothalamic-pituitary-adrenal axis that is not associated with true Cushing’s syndrome (hypercortisolemia) typically seen in a variety of psychiatric states (depression, anxiety), alcoholism, uncontrolled diabetes, and severe obesity. The dexamethasone-CRH stimulation test can be used to help distinguish this disorder from true Cushing’s syndrome. Alternatively, an elevated midnight plasma cortisol level rules out pseudo-Cushing’s because, unlike patients with true Cushing’s syndrome, patients with pseudo-Cushing’s retain the diurnal rhythm of cortisol secretion.