Comprehensive Re-Sequencing of Adrenal Aldosterone Producing Lesions Reveal Three Somatic Mutations near the KCNJ5 Potassium Channel Selectivity Filter

Åkerström, Tobias; Crona, Joakim; Verdugo, Alberto Delgado; Starker, Lee F.; Cupisti, Kenko; Willenberg, Holger S.; Knoefel, Wolfram Trudo; Saeger, Wolfgang; Feller, Alfred C.; Ip, Julian; Soon, Patsy S.; Anlauf, Martin; Alesina, Pier Francesco; Schmid, Kurt Werner; Decaussin, Myriam; Levillain, P.; Wängberg, Bo; Peix, Jean‐Louis; Robinson, Bruce G.; Zedenius, Jan; Bäckdahl, Martin; Caramuta, Stefano; Iwen, K. Alexander; Botling, Johan; Stålberg, Peter; Kraimps, Jean‐Louis; Dralle, Henning; Hellman, Per; Sidhu, Stan; Westin, Gunnar; Lehnert, Hendrik; Walz, Martin K.; Åkerström, Göran; Carling, Tobias; Choi, Murim; Lifton, Richard P.; Björklund, Peyman

doi:10.1371/journal.pone.0041926

Cited by 161 publications

(131 citation statements)

References 29 publications

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“…To assess whether APCCs harbored somatic mutations seen in APA, we developed a custom Ion Torrent AmpliSeq Panel (APA_v1) with 310 multiplexed amplicons targeting the entire coding sequences of genes with described somatic mutations in APA (ATP1A1, ATP2B3, CACNA1D, and KCNJ5) as well as genes shown to harbor germ-line or somatic variants associated with adrenal hyperplasia [phosphodiesterase 11A (PDE11A), phosphodiesterase 8B (PDE8B), and protein kinase, cAMP-dependent, regulatory, type Iα (PRKAR1A)] (7,10,13,(18)(19)(20)(21)(22)(23)(24)(25)(26). NGS was performed on APCCs and paired control adrenal tissue (adjacent ZF and/or medulla), both of which were isolated from formalin-fixed, paraffin-embedded (FFPE) adrenal samples (marked P in Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…The custom Ion AmpliSeq Panel was designed to target genes previously shown to be mutated in APA or other adrenal hyperplasias/ neoplasms (APA_v1 Panel). The APA_v1 Panel contains 310 independent pairs of forward and reverse primers targeting the entire coding regions of ATP1A1, ATB2B3, KCNJ5, and CACNA1D as well as genes shown to harbor germ-line or somatic variants associated with adrenal hyperplasia (PDE11A, PDE8B, and PRKAR1A) (7,10,13,(18)(19)(20)(21)(22)(23)(24)(25)(26). Templates were prepared using the Ion PI Template OT2 200 Kit v2 on the Ion One Touch 2 according to the manufacturer's instructions (Life Technologies).…”

Section: Methodsmentioning

confidence: 99%

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Aldosterone-stimulating somatic gene mutations are common in normal adrenal glands

Nishimoto

Tomlins

Kuick

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

269

273

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Primary aldosteronism (PA) represents the most common cause of secondary hypertension, but little is known regarding its adrenal cellular origins. Recently, aldosterone-producing cell clusters (APCCs) with high expression of aldosterone synthase (CYP11B2) were found in both normal and PA adrenal tissue. PA-causing aldosteroneproducing adenomas (APAs) harbor mutations in genes encoding ion channels/pumps that alter intracellular calcium homeostasis and cause renin-independent aldosterone production through increased CYP11B2 expression. Herein, we hypothesized that APCCs have APArelated aldosterone-stimulating somatic gene mutations. APCCs were studied in 42 normal adrenals from kidney donors. To clarify APCC molecular characteristics, we used microarrays to compare the APCC transcriptome with conventional adrenocortical zones [zona glomerulosa (ZG), zona fasciculata, and zona reticularis]. The APCC transcriptome was most similar to ZG but with an enhanced capacity to produce aldosterone. To determine if APCCs harbored APA-related mutations, we performed targeted next generation sequencing of DNA from 23 APCCs and adjacent normal adrenal tissue isolated from both formalin-fixed, paraffin-embedded, and frozen tissues. Known aldosterone driver mutations were identified in 8 of 23 (35%) APCCs, including mutations in calcium channel, voltage-dependent, L-type, α1D-subunit (CACNA1D; 6 of 23 APCCs) and ATPase, Na + /K + transporting, α1-polypeptide (ATP1A1; 2 of 23 APCCs), which were not observed in the adjacent normal adrenal tissue. Overall, we show three major findings: (i) APCCs are common in normal adrenals, (ii) APCCs harbor somatic mutations known to cause excess aldosterone production, and (iii) the mutation spectrum of aldosteronedriving mutations is different in APCCs from that seen in APA. These results provide molecular support for APCC as a precursor of PA.primary aldosteronism | aldosterone | adrenal | somatic mutations | aldosterone-producing cell cluster P rimary aldosteronism (PA) accounts for 8% of hypertension and is the most common adrenal disease (1-4). PA patients can be classified into those with aldosterone-producing adenomas (APAs), idiopathic hyperaldosteronism, or familial hyperaldosteronism (FH), which is further divided into FH types 1-3 (FHI-FHIII) (5). In 1992, FHI was shown to result from a gene fusion of cytochrome P450, family 11, subfamily B, polypeptide 2 (CYP11B2: aldosterone synthase) and cytochrome P450, family 11, subfamily B, polypeptide 1 (CYP11B1; cortisol synthase) that resulted in zona fasciculata (ZF) expression of CYP11B2 and excess aldosterone production (6). For almost two decades after the original report, no other genetic abnormalities were identified in the other forms of PA.First reported in 2011, exome sequencing identified a series of germ-line and somatic mutations in genes that altered adrenal cell intracellular Ca 2+ in PA. The most common mutations are somatic mutations of the gene encoding the potassium inwardly rectifying channel, subfamily J, member 5 (KCNJ5)...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Aldosterone-stimulating somatic gene mutations are common in normal adrenal glands

Nishimoto

Tomlins

Kuick

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

269

273

View full text Add to dashboard Cite

show abstract

“…Both cortisol-and aldosterone-producing adenomas are more frequent in the left adrenal gland and are more prevalent in females than in males (18), yet the underlying pathophysiology remains unclear. In the case of aldosterone-producing adenomas, KCNJ5 mutations appear to fully account for the female preponderance (25), and this effect is not seen in patients with CACNA1D or ATPase mutations (20,22). In our cohort, there were only three male individuals among the subjects with pure cortisolproducing adenomas, all with CTNNB1 mutations (in one case combined with a GNAS mutation), so it seems possible that a similar gender effect as for KCNJ5 mutations applies to PRKACA and tumors without known mutations.…”

Section: Discussionmentioning

confidence: 99%

PRKACA mutations in cortisol-producing adenomas and adrenal hyperplasia: a single-center study of 60 cases

Thiel

Reis

Haase

et al. 2015

European Journal of Endocrinology

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Objective: Cortisol excess due to adrenal adenomas or hyperplasia causes Cushing's syndrome. Recent genetic studies have identified a somatic PRKACA L206R mutation as a cause of cortisol-producing adenomas. We aimed to compare the clinical features of PRKACA-mutant lesions with those of CTNNB1 mutations, and to search for similar mutations in unilateral hyperplasia or tumors co-secreting aldosterone. Design, patients, and methods: In this study, 60 patients with cortisol excess who had adrenalectomies at our institution between 1992 and 2013 were assessed, and somatic mutations were determined by Sanger sequencing. A total of 36 patients had overt Cushing's syndrome, the remainder were subclinical: 59 cases were adenomas (three bilateral) and one was classified as hyperplasia. Four tumors had proven co-secretion of aldosterone.Results: Among cortisol-secreting unilateral lesions without evidence of co-secretion (nZ52), we identified somatic mutations in PRKACA (L206R) in 23.1%, CTNNB1 (S45P, S45F) in 23.1%, GNAS (R201C) in 5.8%, and CTNNB1CGNAS (S45P, R201H) in 1.9%. PRKACA and GNAS mutations were mutually exclusive. Of the co-secreting tumors, two (50%) had mutations in KCNJ5 (G151R and L168R). The hyperplastic gland showed a PRKACA L206R mutation, while patients with bilateral adenomas did not have known somatic mutations. PRKACA-mutant lesions were associated with younger age, overt Cushing's syndrome, and higher cortisol levels vs non-PRKACA-mutant or CTNNB1-mutant lesions. CTNNB1 mutations were more significantly associated with right than left lesions. Conclusions: PRKACA L206R is present not only in adenomas, but also in unilateral hyperplasia and is associated with more severe autonomous cortisol secretion. Bilateral adenomas may be caused by yet-unknown germline mutations.

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“…10,13,14,[18][19][20][21][22] First, somatic KCNJ5 mutations were more common in female than male patients. Second, in the presence of KCNJ5 mutations, larger tumor size, higher urinary aldosterone, higher aldosterone-renin ratio, and lateralization index were observed, which suggested that excess aldosterone was related to the mutated APA.…”

mentioning

confidence: 99%

“…This finding is in agreement with the result of a previous report. 20 We further investigated the function of the novel somatic KCNJ5 mutation (T148-T149insR). Our findings indicate that this mutation is capable of increasing aldosterone production, as several previously identified mutations (W126R, 23 G151R, 23 T158A, 24 and Ins T149 25 ).…”

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confidence: 99%

Clinical Characteristics of Somatic Mutations in Chinese Patients With Aldosterone-Producing Adenoma

et al. 2015

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Recent studies have shown that somatic mutations in the KCNJ5, ATP1A1, ATP2B3, and CACNA1D genes are associated with the pathogenesis of aldosterone-producing adenoma. Clinical profile and biochemical characteristics of the mutations in Chinese patients with aldosterone-producing adenoma remain unclear. In this study, we performed DNA sequencing in 168 Chinese patients with aldosterone-producing adenoma and found 129 somatic mutations in KCNJ5, 4 in ATP1A1, 1 in ATP2B3, and 1 in CACNA1D. KCNJ5 mutations were more prevalent in female patients and were associated with larger adenomas, higher aldosterone excretion, and lower minimal serum K + concentration. More interestingly, we identified a novel somatic KCNJ5 mutation (c.445-446insGAA, p.T148-T149insR) that could enhance CYP11B2 mRNA upregulation and aldosterone release. This mutation could also cause membrane depolarization and intercellular Ca 2+ increase. In conclusion, somatic KCNJ5 mutations are conspicuously more popular than mutations of other genes in aldosterone-producing adenomas of Chinese patients. The T148-T149insR mutation in KCNJ5 may influence K + channel selectivity and autonomous aldosterone production.

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Comprehensive Re-Sequencing of Adrenal Aldosterone Producing Lesions Reveal Three Somatic Mutations near the KCNJ5 Potassium Channel Selectivity Filter

Cited by 161 publications

References 29 publications

Aldosterone-stimulating somatic gene mutations are common in normal adrenal glands

Aldosterone-stimulating somatic gene mutations are common in normal adrenal glands

PRKACA mutations in cortisol-producing adenomas and adrenal hyperplasia: a single-center study of 60 cases

Clinical Characteristics of Somatic Mutations in Chinese Patients With Aldosterone-Producing Adenoma

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