3 research outputs found
Domains in islet amyloid polypeptide important in fibril formation and toxicity
Islet amyloid deposits are a characteristic pathologic lesion o f the pancreas in type 2
diabetes and are composed primarily of the islet beta cell peptide islet amyloid polypeptide
(IAPP or amylin). Islet amyloid fibrils are toxic to insulin-producing beta cells. Heparan
sulfate proteoglycans (HSPGs) are also a component of islet amyloid in vivo and accelerate
amyloid fibril formation in vitro. Although the cause of amyloid formation in pancreatic
islets is unknown, HSPGs have been proposed to play an initiating role. Impaired processing
of proIAPP, the IAPP precursor, has also been implicated in the mechanism of islet amyloid
formation. The N - and C-terminal cleavage sites at which proIAPP is processed by
prohormone convertases contain series of basic amino acid residues which we hypothesized
may interact with heparan sulfate proteoglycans. We found that a monomeric fragment of
proIAPP (residues 1-30) bound both heparin and heparan sulfate affinity columns.
Substitution of alanine residues for two basic residues in the N-terminal cleavage site
abolished heparin and heparan sulfate binding activity. These data suggest that monomeric
N-terminal human proIAPP contains a heparin-binding domain that is lost during normal
processing of proIAPP.
Although islet amyloid fibrils bind to heparin and heparan sulfate, it is unknown which
residues in IAPP may be responsible for this interaction or whether nonfibrillar (monomeric)
human IAPP also binds to heparin. Three basic residues in human IAPP (Lys1 , A r g 1 1 , and His1 8 ) could potentially contribute to this binding by interacting with negatively charged
sulfate groups in heparin. We found heparin significantly enhanced fibril formation by IAPP
1-37 and 11-37, but not IAPP 12-37, suggesting that A r g 1 1 may be important in the ability of
heparin to enhance IAPP fibril formation. Fibrils generated from IAPP 20-29 (which contains
no basic residues) did not bind heparin, supporting the possibility that specific basic residues
within IAPP may be important in the formation of heparin binding domains within amyloid
fibrils. These findings suggest that basic amino acids may play important roles in IAPP and
amyloid fibril interaction with heparin/heparan sulfate and may contribute to islet amyloid
formation.
A point mutation (serine to glycine at position 20) in IAPP has been identified in a
subpopulation of Japanese patients with a severe, early onset form of type 2 diabetes. We
find S20G IAPP, when applied to INS-1 beta cells in culture, is cytotoxic at lower
concentrations than wild-type IAPP. Using electron microscopy and thioflavin T assays, we
also found that the S20G mutant fibrils form much more rapidly. Increased fibrillogenicity of
S20G IAPP could cause the increased beta cell toxicity of this peptide and may contribute to
early beta cell death and onset of diabetes in patients carrying this mutation.
In conclusion, we speculate that i f N-terminally extended proIAPP is secreted in excessive
amounts from the beta cell in type 2 diabetes, this peptide may be retained by heparan sulfate proteoglycans in the extracellular matrix and form a nidus for subsequent islet amyloid
formation. The possible importance of the three basic residues in mature IAPP in amyloid
fibril formation suggests potential therapeutic targets in preventing islet amyloid formation
and beta cell death in type 2 diabetes. Finally, IAPP toxicity is increased under conditions
that hasten the onset of fibrillogenesis, such as a substitution of glycine for serine at position
20 of the molecule. Domains in IAPP and proIAPP involved in H S PG binding,
fibrillogenesis, and toxicity may make novel therapeutic targets for inhibition o f islet
amyloid formation and the progressive loss of beta cells in type 2 diabetes.Medicine, Faculty ofPathology and Laboratory Medicine, Department ofGraduat
Tracheal Epstein-Barr Virus-Associated Smooth Muscle Tumour in an Hiv-Positive Patient
Epstein-Barr virus-related smooth muscle tumours (EBV-SMTs) are a rare but well recognized non-AIDS-defining malignancy that can also be found in several other immunosuppressed states. Pulmonary involvement of EBVSMTs is not uncommon, but it can present with multifocal lesions in any anatomical site. The present article describes an HIV-positive woman with dyspnea who was found to have a large tracheal EBV-SMT. The authors discuss their approach to diagnosis and management, and present unique follow-up bronchoscopic imaging
Transition from intravenous epoprostenol to oral or subcutaneous therapy in pulmonary arterial hypertension: A retrospective case series and systematic literature review
BACKGROUND: Intravenous epoprostenol, a prostaglandin analogue, has been a mainstay of therapy for patients with advanced pulmonary arterial hypertension (PAH) since the early 1990s. This medication has multiple side effects, and sudden discontinuation is potentially associated with severe sequelae. Several recent case series have described the transition from intravenous to newer oral or subcutaneous therapies. A case series detailing the authors’ experience with such transitions, and a systematic lierature review is presented