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A sensitive and specific diagnosis biomarker, in principle scalable to most cancer types, is needed to reduce the prevalent cancer mortality. Meanwhile, the investigation of diagnosis determinants of a biomarker will facilitate the interpretation of its screening results in clinic. Here we design a large‐scale (1558 enrollments), multicenter (multiple hospitals), and cross‐validation (two datasets) clinic study to validate plasma Hsp90α quantified by  ELISA  as a pan‐cancer biomarker.  ROC  curve shows the optimum diagnostic cutoff is 69.19 ng/ mL  in discriminating various cancer patients from all controls ( AUC  0.895, sensitivity 81.33% and specificity 81.65% in test cohort;  AUC  0.893, sensitivity 81.72% and specificity 81.03% in validation cohort). Similar results are noted in detecting early‐stage cancer patients. Plasma Hsp90α maintains also broad‐spectrum for cancer subtypes, especially with 91.78% sensitivity and 91.96% specificity in patients with  AFP ‐limited liver cancer. In addition, we demonstrate levels of plasma Hsp90α are determined by  ADAM 10 expression, which will affect Hsp90α content in exosomes. Furthermore, Western blotting and  PRM ‐based quantitative proteomics identify that partial false  ELISA ‐negative patients secret high levels of plasma Hsp90α. Mechanism analysis reveal that  TGF β‐ PKC γ gene signature defines a distinct pool of hyperphosphorylated Hsp90α at Theronine residue. In clinic, a mechanistically relevant population of false  ELISA ‐negative patients express also higher levels of  PKC γ. In sum, plasma Hsp90α is a novel pan‐cancer diagnosis biomarker, and cancer diagnosis with plasma Hsp90α is particularly effective in those patients with high expression of  ADAM 10, but may be insufficient to detect the patients with low  ADAM 10 and those with hyperphosphorylated Hsp90α.

A novel pan‐cancer biomarker plasma heat shock protein 90alpha and its diagnosis determinants in clinic