SPIRIT checklist.

(PDF)</p

Dementia international classification of diseases 9^th and 10^th revision (ICD-9 and ICD-10) codes.

Dementia international classification of diseases 9th and 10th revision (ICD-9 and ICD-10) codes.</p

ICD-9 and ICD-10 codes for exclusion criteria reflecting severe mental illness.

ICD-9 and ICD-10 codes for exclusion criteria reflecting severe mental illness.</p

S1 Data -

(XLSX)</p

EHR CDS tool design.

EHR = electronic health records; CDS = clinical decision support.</p

Study protocol documentation.

(PDF)</p

Structural Basis for Resistance to Diverse Classes of NAMPT Inhibitors

<div><p>Inhibiting NAD biosynthesis by blocking the function of nicotinamide phosphoribosyl transferase (NAMPT) is an attractive therapeutic strategy for targeting tumor metabolism. However, the development of drug resistance commonly limits the efficacy of cancer therapeutics. This study identifies mutations in NAMPT that confer resistance to a novel NAMPT inhibitor, GNE-618, in cell culture and <i>in vivo</i>, thus demonstrating that the cytotoxicity of GNE-618 is on target. We determine the crystal structures of six NAMPT mutants in the apo form and in complex with various inhibitors and use cellular, biochemical and structural data to elucidate two resistance mechanisms. One is the surprising finding of allosteric modulation by mutation of residue Ser165, resulting in unwinding of an α-helix that binds the NAMPT substrate 5-phosphoribosyl-1-pyrophosphate (PRPP). The other mechanism is orthosteric blocking of inhibitor binding by mutations of Gly217. Furthermore, by evaluating a panel of diverse small molecule inhibitors, we unravel inhibitor structure activity relationships on the mutant enzymes. These results provide valuable insights into the design of next generation NAMPT inhibitors that offer improved therapeutic potential by evading certain mechanisms of resistance.</p></div

Nampt inhibitors evaluated for potency on mutant NAMPT enzymes.

<p>a) Chemical structures of NAMPT inhibitors, compound 1 is GNE-618, compound 9 is GMX1778, compound 11 is APO866. b) Fold change in IC₅₀ of NAMPT inhibitors on purified mutant NAMPT proteins compared to wild-type enzyme. c) Fold change in IC₅₀ values for S165Y/F mutants compared to wild-type. * fold-change assuming IC₅₀  =  maximum concentration tested (5 µM). Actual fold-change is likely larger than that depicted.</p

Nampt mutations Identified in Resistant Cell Lines.

<p>Cell lines that were selected to grow in the presence of 100 fold the parental IC₅₀ for GNE-618. NA indicates whether the cell line was selected in the presence or absence of 10 uM nicotinic acid. NAPRT1 status did not change as a result of selection for GNE-618 resistance.</p><p>Nampt mutations Identified in Resistant Cell Lines.</p

Crystal structure of NAMPT with resistant mutations mapped.

<p>The NAMPT protein structure is shown in ribbon diagram displaying an active dimer. The monomers of the NAMPT dimer are colored in brown and green, respectively. Two inhibitor molecules of GNE-618 are bound to this structure and are shown as yellow spheres. Some of the resistant mutations are mapped on the structure and are shown as red spheres.</p