Harvey Cushing Treated the First Known Patient With Carney Complex

Figures

• 

  Figure 1.. (A) (top to bottom) Photo of GBS demonstrating his enlarged upper extremities in relation to his chest. Profile of patient GBS with clinical signs of growth hormone excess including prognathism and coarse facial features; also evident are skin tags and freckling, the latter we now realize as part of CNC [7]. (B) Skeleton of patient GBS (left) demonstrating enlarged thorax, extremities, and skull compared with another acromegalic patient from the same collection (right) [7]. (C) (top to bottom) Gross specimen of adrenal gland: Scattered throughout are pigmented nodules consistent with PPNAD. A histological slide demonstrates the intersection of four nodules (80×) [7] (D) (top to bottom) Original sketch from Dr. Cushing’s Brigham notes of the intracranial tumor [6]. Gross specimen of the same pituitary adenoma extending into the right temporal lobe. Histology showed large cells disposed in sheets (600×) [7]. Sampling of the same slice of archived brain tissue today, at our laboratory. (E) (left to right) Dr. Cushing’s sketch of the inferior brain demonstrating the pituitary adenoma in relation to the optic chiasm [6]. Gross specimen from the autopsy demonstrated that the pituitary…

• 

  Figure 2.. Sanger sequencing of the DNA extracted from tissue preserved in 10% buffered neutral formalin since 1914. A guanine-to-adenine missense mutation, leading to p.Arg74His, on exon 3 of the PRKAR1A gene was present in the heterozygous state in germline DNA; loss of heterozygosity is evident in the tumor DNA.

• 

  Figure 3.. p.Arg74His mutation of PRKAR1A alters CREB phosphorylation in basal state and in response to cAMP. The cells bearing the mutation had a higher amount of pCREB than those transfected with WT PRKAR1A after stimulation with cAMP (+cAMP) (P = 0.002); pCREB levels were not significantly different at baseline (–cAMP) (P = 0.18). (A) pCREB (Ser133), CREB, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein levels were quantified by western blot in Hek293 cells after 30-minute induction with 5 µM cAMP or unsupplemented media. Hek293 cells were first transfected (24 hours) with 1 μg WT PRKAR1A, PRKAR1A with R74H mt, or empty vector (EV) plasmid; nontransfected (NT) cells served as an additional control. Transfection experiments were repeated three times with two to three biological replicates per experiment, and western blots for each transfection experiment were repeated twice. (B) Relative pCREB, CREB, and glyceraldehyde 3-phosphate dehydrogenase protein levels were determined by densitometry, and the ratio of phosphorylated to total CREB protein was normalized to the loading control glyceraldehyde 3-phosphate dehydrogenase. Values are mean ± standard error of the mean. *P <…