Low-dose computed tomography (LDCT) Non-Small Cell Lung (NSCLC) screening is associated with high false-positive rates, leading to unnecessary expensive and invasive follow ups. There is a need for minimally invasive approaches to improve the accuracy of NSCLC diagnosis. In addition, NSCLC patients harboring sensitizing mutations in epidermal growth factor receptor EGFR (T790M, L578R) are treated with Osimertinib, a potent tyrosine kinase inhibitor (TKI). However, nearly all patients develop TKI resistance. The underlying mechanisms are not fully understood. Plasma extracellular vesicle (EV) and circulating microRNA (miRNA) have been proposed as biomarkers for cancer screening and to inform treatment decisions. However, the identification of highly sensitive and broadly predictive core miRNA signatures remains a challenge. Also, how these systemic and diverse miRNAs impact cancer drug response is not well understood. Using an integrative approach, we examined plasma EV and circulating miRNA isolated from NSCLC patients versus screening controls with a similar risk profile. We found that combining EV (Hsa-miR-184, Let-7b-5p) and circulating (Hsa-miR-22-3p) miRNAs abundance robustly discriminates between NSCLC patients and high-risk cancer-free controls. Further, we found that Hsa-miR-22-3p, Hsa-miR-184, and Let-7b-5p functionally converge on WNT/βcatenin and mTOR/AKT signaling axes, known cancer therapy resistance signals. Targeting Hsa-miR-22-3p and Hsa-miR-184 desensitized EGFR-mutated (T790M, L578R) NSCLC cells to Osimertinib. These findings suggest that the expression levels of circulating hsa-miR-22-3p combined with EV hsa-miR-184 and Let-7b-5p levels potentially define a core biomarker signature for improving the accuracy of NSCLC diagnosis. Importantly, these biomarkers have the potential to enable prospective identification of patients who are at risk of responding poorly to Osimertinib alone but likely to benefit from Osimertinib/AKT blockade combination treatments.

How organisms control organ size is not fully understood. We found that Syd/JIP3 is required for proper wing size in Drosophila. JIP3 mutations are associated with organ size defects in mammals.Our mechanistic studies place Syd/JIP3 downstream of Hippo. Syd/JIP3 is dispensable for Hippo transcriptional activity but it is essential for Diap1 protein stability and function. Consistent with this observation, Syd/JIP3 inhibition reduces Diap1 protein levels and, correspondingly, elevates cell death in wing tissues. Partial restoration of Diap1 rescues wing size. In addition, Syd/JIP3 is upregulated in response to Yorkie activation. Importantly, Syd/JIP3 is required for the full effect of Yorkie-mediated tissue overgrowth. Thus, Syd/JIP3 is a novel organ size regulator in the Hippo pathway. This study provides mechanistic insights into the recent and perplexing link between JIP3 mutations and organ size defects in mammals, including in humans where de novo JIP3 variants are associated with microcephaly..

Highlights• Syd/JIP3 is required for proper Drosophila wing size• Syd/JIP3 functions downstream of Yorkie by stabilizing Diap1 to promote cell survival during rapid tissue growth • Yorkie-mediated tissue growth is highly sensitive to Syd/JIP3 dosage • Syd/JIP3 is downregulated at the end of the tissue expansion phase AbstractHow organisms control organ size is not fully understood. We found that Syd/JIP3 is required for proper wing size in Drosophila. JIP3 mutations are associated with organ size defects in mammals.Our mechanistic studies place Syd/JIP3 downstream of Hippo. Syd/JIP3 is dispensable for Hippo transcriptional activity but it is essential for Diap1 protein stability and function. Consistent with this observation, Syd/JIP3 inhibition reduces Diap1 protein levels and, correspondingly, elevates cell death in wing tissues. Partial restoration of Diap1 rescues wing size. In addition, Syd/JIP3 is upregulated in response to Yorkie activation. Importantly, Syd/JIP3 is required for the full effect of Yorkie-mediated tissue overgrowth. Thus, Syd/JIP3 is a novel organ size regulator in the Hippo pathway. This study provides mechanistic insights into the recent and perplexing link between JIP3 mutations and organ size defects in mammals, including in humans where de novo JIP3 variants are associated with microcephaly.