Supplementary Figure S3 from Anti-CLL1 Chimeric Antigen Receptor T-Cell Therapy in Children with Relapsed/Refractory Acute Myeloid Leukemia

Supplementary Figure S3. Cytokine profiles of enrolled AML cases treated with CLL1 CAR-T cells. Serum INFγ (A), IL-2 (B), IL-4 (C), IL-10 (D), and TNFα (E) (yellow shaded area represents the reference value) were evaluated using AimPlex BioSciences Human Premixed Multiplex Kits (red line with solid circle, patient 1; blue line with solid square, patient 2; dark green with solid circle, patient 3; purple line with empty square, patient 4).</p

Supplementary Figure S1 from Anti-CLL1 Chimeric Antigen Receptor T-Cell Therapy in Children with Relapsed/Refractory Acute Myeloid Leukemia

Supplementary Figure S1. The flow-chart of CALSIII-AML 18 and our study.</p

Supplementary Figure S2 from Anti-CLL1 Chimeric Antigen Receptor T-Cell Therapy in Children with Relapsed/Refractory Acute Myeloid Leukemia

Supplementary Figure S2. The lymphocyte population before anti-CLL1 CAR-T cell therapy among these four R/R-AML patients.</p

Supplementary Figure S4 from Anti-CLL1 Chimeric Antigen Receptor T-Cell Therapy in Children with Relapsed/Refractory Acute Myeloid Leukemia

Supplementary FigureS4. Suggested optional approaches for the management of children with R/R-AML.</p

Additional file 7 of CircRNF220, not its linear cognate gene RNF220, regulates cell growth and is associated with relapse in pediatric acute myeloid leukemia

Additional file 7: Supplementary Table S3. Sequences of the PCR primers in this study (5’-3’)

Additional file 3 of CircRNF220, not its linear cognate gene RNF220, regulates cell growth and is associated with relapse in pediatric acute myeloid leukemia

Additional file 3: Supplementary Fig S3. Relative level of circRNF220 in AML cells lentiviral infected with circRNF220 shRNA or circRNF220 overexpression vector. (A) Schematic of the circRNF220 overexpression vector. (B) Efficient overexpression of circRNF220 in HL-60, THP-1 and K562 cell lines. (C) circRNF220 overexpression vector did not influence parental gene expression in HL-60, THP-1 and K562 cell lines. (D) Schematic representation of the sites of the siRNA specific to the back-splice junction of circRNF220. (E) Efficient knockdown of circRNF220 in AML-K233, AML-K262, and AML-K450 three patients’ cells. (F) Specific siRNA-circRNF220 did not influence parental gene expression in AML-K233, AML-K262, and AML-K450 three patients’ cells. (G) Efficient knockdown of circRNF220 in AML-A750, AML-943, and AML-E85 three patients’ cells. (H) Specific siRNA-circRNF220 did not influence parental gene expression in AML-A750, AML-943, and AML-E85 three patients’ cells. *P < 0.05, **P < 0.01, ***P < 0.001. ns: not significant

Additional file 6 of CircRNF220, not its linear cognate gene RNF220, regulates cell growth and is associated with relapse in pediatric acute myeloid leukemia

Additional file 6: Supplementary Table S2. Detailed characteristics of pediatric patients with newly diagnosed Hematologic Disorders except for AML (n=147)

DataSheet1_Effects of NRAS Mutations on Leukemogenesis and Targeting of Children With Acute Lymphoblastic Leukemia.pdf

Through the advancements in recent decades, childhood acute lymphoblastic leukemia (ALL) is gradually becoming a highly curable disease. However, the truth is there remaining relapse in ∼15% of ALL cases with dismal outcomes. RAS mutations, in particular NRAS mutations, were predominant mutations affecting relapse susceptibility. KRAS mutations targeting has been successfully exploited, while NRAS mutation targeting remains to be explored due to its complicated and compensatory mechanisms. Using targeted sequencing, we profiled RAS mutations in 333 primary and 18 relapsed ALL patients and examined their impact on ALL leukemogenesis, therapeutic potential, and treatment outcome. Cumulative analysis showed that RAS mutations were associated with a higher relapse incidence in children with ALL. In vitro cellular assays revealed that about one-third of the NRAS mutations significantly transformed Ba/F3 cells as measured by IL3-independent growth. Meanwhile, we applied a high-throughput drug screening method to characterize variable mutation-related candidate targeted agents and uncovered that leukemogenic-NRAS mutations might respond to MEK, autophagy, Akt, EGFR signaling, Polo−like Kinase, Src signaling, and TGF−β receptor inhibition depending on the mutation profile.</p

Additional file 2 of CircRNF220, not its linear cognate gene RNF220, regulates cell growth and is associated with relapse in pediatric acute myeloid leukemia

Additional file 2: Supplementary Fig S2. ROC analysis for circRNF220 in bone marrow of AML patients. (A) The AUC indistinguishing AML and normal individual was 0.9735. (B) The AUC indistinguishing AML and ALL was 0.9209. (C) The AUC indistinguishing AML and other hematologic malignancies was 0.9274. (D)Spearman’s coefficient scatters plot of the fold-changes of circRNF220 in the BM and PB samples (r=0.8107, P<0.0001). BM: bone marrow, PB: peripheral blood

Additional file 4 of CircRNF220, not its linear cognate gene RNF220, regulates cell growth and is associated with relapse in pediatric acute myeloid leukemia

Additional file 4: Supplementary Fig S4. CircRNF220 regulates AML proliferation and apoptosis through targeting miR-30a. (A) Correlation between circRNF220 and miR-30b, miR-30c, miR-30d, miR-30e, respectively. (B) The miR-30 family expression levels in overexpression circRNF220 by real-time PCR in HL-60. (C) RNA pulldown using biotinylated circRNF220 and detection of miR-30a in HL-60. (D) miR-30 family expression levels in AML cells transfected with miR-30a. (E) Correlation between miR-30a and MYSM1, GLUD2, ELK3, and IER2, respectively. (F) Correlation between circRNF220 and MYSM1, GLUD2, ELK3, and IER2, respectively. (G) Cell proliferation increased upon inhibitor-miR-30a and was impaired after circRNF220 KD in HL-60 cells. (H) Apoptosis was arrested by miR-30a inhibitor and which effect was eliminated after circRNF220 KD in HL-60 cells. The numbers indicate the percentages of apoptotic cells. The data are presented as the mean values of triplicates. (I) The relative expression of circRNF220 and miR-30a in AML primary cells when transfected different small interfering RNA. (J) Apoptosis was arrested upon miR-30a inhibitor treatment, and this effect was eliminated after circRNF220 siRNA transfection in primary pediatric AML cells. The numbers indicate the percentages of apoptotic primary AML cells. The data are presented as the mean values of triplicates. *P < 0.05, **P < 0.01, ***P < 0.001. ns: not significant