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Primary Pigmented Nodular Adrenocortical Disease via the PDE11A Gene

  • Summary and Pricing
  • Clinical Features and Genetics
  • Citations
  • Methods
  • Ordering/Specimens
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TEST METHODS

Sequencing

Test Code Test Copy GenesIndividual Gene PriceCPT Code Copy CPT Codes
1292 PDE11A$1250.00 81479 Add to Order
Targeted Testing

For ordering targeted known variants, please proceed to our Targeted Variants landing page.

Turnaround Time

The great majority of tests are completed within 18 days.

Clinical Sensitivity
The clinical sensitivity of PDE11A germline mutations in patients with PPNAD is currently unknown as only a small number of individual cases have been reported.

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Clinical Features
Primary pigmented nodular adrenocortical disease (PPNAD) is a rare disorder that causes adrenocorticotropic hormone (ATCH)-independent adrenal hyperplasia resulting in Cushing Syndrome (Manipadam et al. 2011). Cushing syndrome causes the body to have prolonged exposure to cortisol leading to obesity, severe fatigue, weak muscles, high blood pressure, high blood sugar, irritability, and anxiety. Cushing syndrome has a prevalence of 1/26,000, and PPNAD represents 2% of these cases (http://www.orpha.net/). PPNAD is often associated with Carney's complex, however, isolated PPNAD is also known to occur less frequently (Stratakis 2009). In PPNAD, the adrenal glands are small to normal sized and harbor multiple small cortical pigmented nodules. Most cases of PPNAD occur in the second and third decades of life, but it can also occur very early (e.g. 2-3 years of age), and it appears to be more frequent in females (Almeida and Stratakis 2010).
Genetics
Primary pigmented nodular adrenocortical disease is inherited in an autosomal dominant manner and can be caused by germline mutations in the PRKAR1A and PDE11A genes (Gaillard et al. 2010). The PRKAR1A gene encodes a tumor suppressor that acts as a cyclic-AMP-dependent signaling molecule. The signaling molecule phosphorylates many downstream targets that are involved in transcription, metabolism, cell cycle progression and apoptosis (Rothenbuhler and Stratakis 2010). PDE11A catalyzes the hydrolysis of cAMP and cGMP and is expressed in several endocrine tissues (Almeida and Stratakis 2010).  Both PDE11A and PRKAR1A repress the activity of protein kinase A, which is involved in cellular proliferation, and both may act in concert, when mutated, to promote adrenal cortical cell proliferation (Bar-Lev and Annes 2012).
Testing Strategy
The dual 3', 5'-cyclic-AMP and -GMP phosphodiesterase 11A is encoded by 20 exons from the PDE11A gene on chromosome 2q31.2. Testing is accomplished by amplifying each coding exon and ~20 bp of adjacent noncoding sequence, then determining the nucleotide sequence using standard Sanger dideoxy sequencing methods and a capillary electrophoresis instrument. We will also sequence any single exon (Test #100) in family members of patients with a known mutation or to confirm research results.
Indications for Test
Individuals with a clinical presentation of Primary Pigmented Nodular Adrenocortical Disease and individuals with a family history of disease.

Gene

Official Gene Symbol OMIM ID
PDE11A 604961
Inheritance Abbreviation
Autosomal Dominant AD
Autosomal Recessive AR
X-Linked XL
Mitochondrial MT

Related Tests

Name
Carney Complex (CNC) via the PRKAR1A Gene
Primary Macronodular Adrenal Hyperplasia via the ARMC5 Gene

CONTACTS

Genetic Counselors
Geneticist
Citations
  • Almeida MQ, Stratakis CA. 2010. Carney complex and other conditions associated with micronodular adrenal hyperplasias. Best Practice & Research Clinical Endocrinology & Metabolism 24: 907–914. PubMed ID: 21115159
  • Bar-Lev A, Annes JP. 2012. Genetics of adrenocortical disease: an update. Current Opinion in Endocrinology & Diabetes and Obesity 19: 159–167. PubMed ID: 22476103
  • Gaillard RC, Bertherat J, Carney JA. 2010. Familial micronodular adrenocortical disease, Cushing syndrome, and mutations of the gene encoding phosphodiesterase 11A4 (PDE11A). PubMed ID: 20351491
  • Manipadam MT, Abraham R, Sen S, Simon A. 2011. Primary pigmented nodular adrenocortical disease. J Indian Assoc Pediatr Surg 16: 160–162. PubMed ID: 22121318
  • Orphanet
  • Rothenbuhler A, Stratakis CA. 2010. Clinical and molecular genetics of Carney complex. Best Practice & Research Clinical Endocrinology & Metabolism 24: 389–399. PubMed ID: 20833331
  • Stratakis CA. 2009. New genes and/or molecular pathways associated with adrenal hyperplasias and related adrenocortical tumors. Molecular and Cellular Endocrinology 300: 152–157. PubMed ID: 19063937
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TEST METHODS

Bi-Directional Sanger Sequencing

Test Procedure

Nomenclature for sequence variants was from the Human Genome Variation Society (http://www.hgvs.org).  As required, DNA is extracted from the patient specimen.  PCR is used to amplify the indicated exons plus additional flanking non-coding sequence.  After cleaning of the PCR products, cycle sequencing is carried out using the ABI Big Dye Terminator v.3.0 kit.  Products are resolved by electrophoresis on an ABI 3730xl capillary sequencer.  In most cases, sequencing is performed in both forward and reverse directions; in some cases, sequencing is performed twice in either the forward or reverse directions.  In nearly all cases, the full coding region of each exon as well as 20 bases of non-coding DNA flanking the exon are sequenced.

Analytical Validity

As of March 2016, we compared 17.37 Mb of Sanger DNA sequence generated at PreventionGenetics to NextGen sequence generated in other labs. We detected only 4 errors in our Sanger sequences, and these were all due to allele dropout during PCR. For Proficiency Testing, both external and internal, in the 12 years of our lab operation we have Sanger sequenced roughly 8,800 PCR amplicons. Only one error has been identified, and this was due to sequence analysis error.

Our Sanger sequencing is capable of detecting virtually all nucleotide substitutions within the PCR amplicons. Similarly, we detect essentially all heterozygous or homozygous deletions within the amplicons. Homozygous deletions which overlap one or more PCR primer annealing sites are detectable as PCR failure. Heterozygous deletions which overlap one or more PCR primer annealing sites are usually not detected (see Analytical Limitations). All heterozygous insertions within the amplicons up to about 100 nucleotides in length appear to be detectable. Larger heterozygous insertions may not be detected. All homozygous insertions within the amplicons up to about 300 nucleotides in length appear to be detectable. Larger homozygous insertions may masquerade as homozygous deletions (PCR failure).

Analytical Limitations

In exons where our sequencing did not reveal any variation between the two alleles, we cannot be certain that we were able to PCR amplify both of the patient’s alleles. Occasionally, a patient may carry an allele which does not amplify, due for example to a deletion or a large insertion. In these cases, the report contains no information about the second allele.

Similarly, our sequencing tests have almost no power to detect duplications, triplications, etc. of the gene sequences.

In most cases, only the indicated exons and roughly 20 bp of flanking non-coding sequence on each side are analyzed. Test reports contain little or no information about other portions of the gene, including many regulatory regions.

In nearly all cases, we are unable to determine the phase of sequence variants. In particular, when we find two likely causative mutations for recessive disorders, we cannot be certain that the mutations are on different alleles.

Our ability to detect minor sequence variants, due for example to somatic mosaicism is limited. Sequence variants that are present in less than 50% of the patient’s nucleated cells may not be detected.

Runs of mononucleotide repeats (eg (A)n or (T)n) with n >8 in the reference sequence are generally not analyzed because of strand slippage during PCR and cycle sequencing.

Unless otherwise indicated, the sequence data that we report are based on DNA isolated from a specific tissue (usually leukocytes). Test reports contain no information about gene sequences in other tissues.

Order Kits

Ordering Options


myPrevent - Online Ordering
  • The test can be added to your online orders in the Summary and Pricing section.
  • Once the test has been added log in to myPrevent to fill out an online requisition form.
REQUISITION FORM
  • A completed requisition form must accompany all specimens.
  • Billing information along with specimen and shipping instructions are within the requisition form.
  • All testing must be ordered by a qualified healthcare provider.

SPECIMEN TYPES
WHOLE BLOOD

(Delivery accepted Monday - Saturday)

  • Collect 3 ml -5 ml (5 ml preferred) of whole blood in EDTA (purple top tube) or ACD (yellow top tube). For Test #500-DNA Banking only, collect 10 ml -20 ml of whole blood.
  • For small babies, we require a minimum of 1 ml of blood.
  • Only one blood tube is required for multiple tests.
  • Ship blood tubes at room temperature in an insulated container. Do not freeze blood.
  • During hot weather, include a frozen ice pack in the shipping container. Place a paper towel or other thin material between the ice pack and the blood tube.
  • In cold weather, include an unfrozen ice pack in the shipping container as insulation.
  • At room temperature, blood specimen is stable for up to 48 hours.
  • If refrigerated, blood specimen is stable for up to one week.
  • Label the tube with the patient name, date of birth and/or ID number.

DNA

(Delivery accepted Monday - Saturday)

  • Send in screw cap tube at least 5 µg -10 µg of purified DNA at a concentration of at least 20 µg/ml for NGS and Sanger tests and at least 5 µg of purified DNA at a concentration of at least 100 µg/ml for gene-centric aCGH, MLPA, and CMA tests, minimum 2 µg for limited specimens.
  • For requests requiring more than one test, send an additional 5 µg DNA per test ordered when possible.
  • DNA may be shipped at room temperature.
  • Label the tube with the composition of the solute, DNA concentration as well as the patient’s name, date of birth, and/or ID number.
  • We only accept genomic DNA for testing. We do NOT accept products of whole genome amplification reactions or other amplification reactions.

CELL CULTURE

(Delivery preferred Monday - Thursday)

  • PreventionGenetics should be notified in advance of arrival of a cell culture.
  • Culture and send at least two T25 flasks of confluent cells.
  • Some panels may require additional flasks (dependent on size of genes, amount of Sanger sequencing required, etc.). Multiple test requests may also require additional flasks. Please contact us for details.
  • Send specimens in insulated, shatterproof container overnight.
  • Cell cultures may be shipped at room temperature or refrigerated.
  • Label the flasks with the patient name, date of birth, and/or ID number.
  • We strongly recommend maintaining a local back-up culture. We do not culture cells.
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