The incidence of early onset colorectal cancer (EoCRC), defined as colorectal cancer (CRC) in patients under age 50, has been increasing in the United States. This is due to a birth cohort effect in which the younger generation has experienced an accelerating rise in EoCRC for reasons currently unknown, although epidemiologic research points to several traditional and emerging generation-specific risk fac-tors. There are several racial/ethnic and geographic differences in the presentation of EoCRC with dis-parate outcomes. A subset of EoCRC patients have a familial or hereditary cause of EoCRC, although the etiology for most EoCRC remains to be discovered. Our current approach to prevention and early detection includes early screening for familial CRC, germline genetic testing for all cases of EoCRC, tri -age of alarm symptoms with prompt evaluation of red flag signs and symptoms (such as hematochezia, iron deficiency anemia, and unexplained weight loss), offering a menu of average-risk screening options to those age 45 and older, and performing outreach/navigation to improve opportunistic screening uptake. Unfortunately, full actualization of these approaches remains suboptimal, and the increasing burden of EoCRC demands immediate action. Opportunities to improve prevention and early detection of EoCRC include initiating organized screening approaches through leveraging the electronic health record, centralization of care in medical homes, outreach using blockchain or social media technology, and biotechnological innovations in diagnosis and risk stratification.

AVI file - 8883KB, siRNA Control cells progressing through mitosis after MMC treatment.

Supplementary Video 2 from Abnormal Cytokinesis after X-Irradiation in Tumor Cells that Override the G2 DNA Damage Checkpoint

PDF file - 64KB, Class of drugs used in the drug sensitivity assays performed in MiaPaCa2 cells against siRNA Control and HuR, respective IC50 of each drug identified.

PDF file - 471KB, HuR silencing induced DNA damage breaks in PL5 cells.

PDF file - 395K, A) Binding assays of H1299 cells transfected with HA-tagged full-length and truncated HSP70 encoding the amino acids (aa) shown, treated with 20 uM biotinylated PES (B-PES) and 20 uM PES-Cl (PES-Cl). Biotinylated PES complexes were precipitated with avidin beads, eluted and detected with an anti-HA antibody following imunoblotting. Input is shown on the top panel, immunoprecipitation (IP) with avidin is depicted on the bottom panel. B) Cell viability of WI38 non-transformed fibroblasts after 48 hour treatment with 10 uM PES and PES-Cl. The values shown are the averaged results from three independent experiments, and error bars mark standard deviations. C) Western blot analysis of HSP90 client proteins in soluble cell extract following 48 hours of treatment with 10 uM PES-Cl. Actin is shown as a loading control. D) In silico docking of PES-Cl to the substrate binding domain of HSP70.

Supplementary Table S1: Proteins upregulated follwoing vemurafenib treatment of SK-MEL-28VR1 cells. Supplementary Figure S1: PHGDH gene ablation of SK-MEL-28VR1 cells. Supplementary Figure S2: Gemcitabine induced sensitization of SK-MEL-28VR1 and BxPC3M1 cells to vemurafenib. Supplementary Table S2: Combination index values for gemcitabine+vemurafenib combination in SK-MEL-28VR1 cells. Supplementary Table S3: Combination index values for gemcitabine+vemurafenib combination in BxPC3M1 cells.

PDF file - 280K, A) Chemical structures of PES and synthesized derivatives. P=pyrrolidine; CM= carboxymethyl; Cl=chloride. B) MTT cell viability analyses of H1299 and PANC1 cells after 48 hour treatment with 10 uM PES, PES-P, PES-PCM and PES-PCl. The values shown are the averaged results from three independent experiments, and error bars mark standard deviations. C) Immunoblot analysis of H1299 cells following treatment with 10 uM of the compounds indicated for the timepoints indicated, using antisera to the autophagy (p62SQSTM1 and LC3) and apoptosis (cleaved lamin A) markers indicated, as well as HSP70 and actin (control). The arrows mark monomeric and oligomeric forms of p62SQSTM1, as well as cleaved and lipidated forms of LC3 I and II.

Supplementary Figure 1. Hedgehog pathway is not activated in U251 cells. Supplementary Figure 2. Colony formation from passaged U251 cells after Nestin knockdown. Supplementary Figure 3. GBM cells from PDX undergo apoptosis after Nestin knockdown. Supplementary Figure 4. Nestin deletion causes G2/M arrest and apoptosis in U251 cells. Supplementary Figure 5. Colony formation from U87 cells after transduction with Nestin shRNA. Supplementary Figure 6. No alterations in the mRNA expression of TUBB2A and TUBB2B in U251 cells after Nestin manipulation. Supplementary Figure 7. No alterations in the expression of Nestin mRNA and protein in U251 cells after βII-tubulin knockdown.

PDF file - 497KB, HuR regulates WEE1 expression.

PDF file - 130K, A) Chemical structure of biotinylated PES B) Chemical structure of PES and derivatives NSC85560, NSC166772 and NSC303578 (nomenclature is from the National Cancer Institute Drug Repository). C) Cell viability of H1299 and PANC1 cells after 48 hour treatment with 10 uM PES, NSC85560 (855600), NSC166777 (16677) and NSC303578 (303578). Data are the averaged results from three independent experiments, and error bars mark standard deviations. D) Immunoblot analysis of H1299 cells following treatment with 10 uM of the compounds indicated for the timepoints indicated, using antisera to the autophagy (p62SQSTM1 and LC3) and apoptosis (cleaved lamin A) markers indicated, as well as HSP70 and actin (control). The arrows mark monomeric and oligomeric forms of p62SQSTM1, as well as cleaved and lipidated forms of LC3 I and II.

AVI file - 8882KB, Untreated siRNA HuR cells progressing through mitosis.

PDF file - 2204KB, Different DNA damaging agents activate HuR.