Journal: Molecular cancer therapeutics

Article Title: ERADICATION OF HETEROGENEOUS TUMORS BY T-CELLS TARGETED WITH COMBINATION BISPECIFIC CHEMICALLY SELF-ASSEMBLED NANORINGS (CSANs)

doi: 10.1158/1535-7163.MCT-22-0515

Figure Lengend Snippet: Target HT-29 cell (CD133 high EpCAM high colon carcinoma) lysis, by unactivated PBMCs, was evaluated at a set 10:1 E:T ratio with increasing concentrations of (A) αCD133/αCD3 CSANs, (B) αEpCAM/αCD3 CSANs or (C) both αCD133/αCD3 CSANs and αEpCAM/αCD3 CSANs in combination. Target MDA-MB-231 cell (CD133 low EpCAM low TNBC cells) lysis, by unactivated PBMCs, was evaluated at a set 10:1 E:T ratio with increasing concentrations of (D) αCD133/αCD3 CSANs and (E) αEpCAM/αCD3 CSANs, as well as in (F) combination. When treated in combination, both αCD133/αCD3 CSANs and αEpCAM/αCD3 CSANs were used at a 1:1 ratio. CSAN controls were generated by combining a targeted DHFR 2 with a non-targeted DHFR 2 (1DD). Data shown was obtained from one donor (n=3), but representative of three donors. *P<0.05 with respect to targeted CSAN therapy and non-targeted CD3 control, by 2-tailed Student’s t test.

Article Snippet: HT-29 and MDA-MB-231 cells were obtained from American Type Culture Collection (ATCC, Rockville, MD) and were monolayer cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and L-glutamine at 37°C in 5% CO2.

Techniques: Labeling, Lysis, Generated, Control

Journal: Molecular cancer therapeutics

Article Title: ERADICATION OF HETEROGENEOUS TUMORS BY T-CELLS TARGETED WITH COMBINATION BISPECIFIC CHEMICALLY SELF-ASSEMBLED NANORINGS (CSANs)

doi: 10.1158/1535-7163.MCT-22-0515

Figure Lengend Snippet: Unactivated PBMCs were co-cultured with media, 100 nM 1DD/αCD3 CSANS, 100 nM αCD133/αCD3 CSANs, 100 nM αEpCAM/αCD3 CSANs, or both 50 nM αCD133/αCD3 and 50 nM αEpCAM/αCD3 CSANs to generate non-labeled PBMCS, non-targeted CSANs, αCD133 CSANs, αEpCAM CSANs or dual targeting αCD133/αEpCAM CSANs, respectively. Generated constructs were incubated in the presence of no target cells, U87-MG cells (EpCAM neg CD133 neg ), MCF-7 cells (EpCAM high CD133 neg ), HT-29 cells (EpCAM high CD133 pos ), or MDA-MB-231 cells (EpCAM low CD133 partial pos ). Following the 24-hour incubation, the media was analyzed for (A) IL-2 and (B) IFN-γ. Data shown was obtained from one donor (n=3), but representative of three donors. *P<0.05, by 2-tailed Student’s t test and one-way ANOVA.

Article Snippet: HT-29 and MDA-MB-231 cells were obtained from American Type Culture Collection (ATCC, Rockville, MD) and were monolayer cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and L-glutamine at 37°C in 5% CO2.

Techniques: Cell Culture, Labeling, Generated, Construct, Incubation

Journal: Molecular cancer therapeutics

Article Title: ERADICATION OF HETEROGENEOUS TUMORS BY T-CELLS TARGETED WITH COMBINATION BISPECIFIC CHEMICALLY SELF-ASSEMBLED NANORINGS (CSANs)

doi: 10.1158/1535-7163.MCT-22-0515

Figure Lengend Snippet: NSG mice were inoculated in the flank with 1.0×10 6 HT-29 cells. Cohorts were randomized when tumors were ~80 mm 3 and IV inoculated with 20 million PBMCs. Treatments were initiated 4 days later, including: PBS, PBMC only (this experimental group acquired treatments of PBS only following the initial PBMC engraftment), 1 mg/kg αCD3 monospecific CSANs, 1 mg/kg αCD133 monospecific CSANs, and 1 mg/kg αEpCAM/αCD3 bispecific CSANs (n=5). Treatments were administered every 2 days for a total of 5 treatments. (A) Tumor growth was monitored every by caliper and recorded as mm 3 . *P<0.05 with respect to readings statistically significant from the 1DD/αCD3 CSAN control group, by 2-tailed Student’s t test. (B) Survival was monitored out to 85 days when graft vs host (GVH) disease symptoms were no longer manageable. Kaplan-Mier Survival Plot. *P<0.0001 with respect to PBS control, by log rank test. All in vivo experiments were performed independently and at least twice.

Article Snippet: HT-29 and MDA-MB-231 cells were obtained from American Type Culture Collection (ATCC, Rockville, MD) and were monolayer cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, and L-glutamine at 37°C in 5% CO2.

Techniques: In Vivo, Control

Journal: British Journal of Nutrition

Article Title: Effect of β-carotene-rich tomato lycopene β-cyclase (tlcy-b) on cell growth inhibition in HT-29 colon adenocarcinoma cells

doi: 10.1017/s0007114508169902

Figure Lengend Snippet: Fig. 1. Effects of tomato lycopene b-cyclase (tcly-b) tomato digestate on the growth of HT-29 cells. (A) Effects of different digestate concentrations for 24 h: (A), control; (B), digestate at 20 ml/l; (o), digestate at 50 ml/l; ( ), digestate at 100 ml/l. Values are the means of three different experiments, with standard errors represented by vertical bars. a,b,c Mean values with unlike letters were significantly different (P,0·05; Fischer’s test). (B) Effects of different times (24, 48, 72 h) of treatment: (A), control; ( ), digestate at 100 ml/l. Values are the means of three different experiments, with standard errors represented by vertical bars. a,b,c,d Mean values with unlike letters were significantly different (P,0·002; Tukey’s test). The treatment–time interaction was significant (P,0·05).

Article Snippet: HT-29 human colon adenocarcinoma cells (American Type Culture Collection (ATCC), Rockville, MD, USA) were grown in minimum essential medium.

Techniques: Control

Journal: British Journal of Nutrition

Article Title: Effect of β-carotene-rich tomato lycopene β-cyclase (tlcy-b) on cell growth inhibition in HT-29 colon adenocarcinoma cells

doi: 10.1017/s0007114508169902

Figure Lengend Snippet: Fig. 2. Effects of varying tomato lycopene b-cyclase (tcly-b) tomato digestate concentration on apoptosis induction in HT-29 cells: (A), control; (B), diges- tate at 20 ml/l; (o), digestate at 50 ml/l; ( ), digestate at 100 ml/l. (A) Percen- tage of terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL)-positive cells. (B) Caspase-3 activation. Cells were treated for 24 h. Values are the means of three different experiments, with standard errors represented by vertical bars. a,b,c Mean values with unlike letters were significantly different (P,0·05; Fischer’s test).

Article Snippet: HT-29 human colon adenocarcinoma cells (American Type Culture Collection (ATCC), Rockville, MD, USA) were grown in minimum essential medium.

Techniques: Concentration Assay, Control, TUNEL Assay, Activation Assay

Journal: British Journal of Nutrition

Article Title: Effect of β-carotene-rich tomato lycopene β-cyclase (tlcy-b) on cell growth inhibition in HT-29 colon adenocarcinoma cells

doi: 10.1017/s0007114508169902

Figure Lengend Snippet: Fig. 3. Bax, Bcl-2 and Bcl-xl expression in HT-29 cells treated with varying tomato lycopene b-cyclase(tcly-b) tomato digestate concentrations for 24 h: (A), control (C); (B), digestate at 20 ml/l (T20); (o), digestate at 50 ml/l (T50); ( ), digestate at 100 ml/l (T100). (A) Representative Western blot analyses. (B) Bcl-2 densitometric analysis. (C) Bcl-xl densitometric analysis. Values are the means of three different determinations, with standard errors represented by vertical bars. a,b,c Mean values with unlike letters were significantly different (P,0·05; Fischer’s test).

Article Snippet: HT-29 human colon adenocarcinoma cells (American Type Culture Collection (ATCC), Rockville, MD, USA) were grown in minimum essential medium.

Techniques: Expressing, Control, Western Blot

Journal: British Journal of Nutrition

Article Title: Effect of β-carotene-rich tomato lycopene β-cyclase (tlcy-b) on cell growth inhibition in HT-29 colon adenocarcinoma cells

doi: 10.1017/s0007114508169902

Figure Lengend Snippet: Fig. 6. b-Carotene incorporation and/or association in HT-29 cells treated with tomato lycopene b-cyclase (tcly-b) tomato digestate (B) and purified b-carotene (X) for 24 h. The concentration of digestate in cultured medium was 100 ml/l and that of the purified carotenoid was 0·8 mM. Values are the means of three different experiments, with standard errors represented by vertical bars. * Mean value was significantly different from that of the digestate-treated cells (P,0·005; Tukey’s test).

Article Snippet: HT-29 human colon adenocarcinoma cells (American Type Culture Collection (ATCC), Rockville, MD, USA) were grown in minimum essential medium.

Techniques: Concentration Assay, Cell Culture

Journal: British Journal of Nutrition

Article Title: Effect of β-carotene-rich tomato lycopene β-cyclase (tlcy-b) on cell growth inhibition in HT-29 colon adenocarcinoma cells

doi: 10.1017/s0007114508169902

Figure Lengend Snippet: Fig. 5. Comparison of b-carotene (X) and tomato lycopene b-cyclase (tcly-b) tomato digestate (B) ability in inhibiting HT-29 cells growth after 24 h treat- ment. Concentrations of tcly-b tomato digestate of 20, 50 and 100 ml/l corre- spond to b-carotene concentrations of 0·08, 0·16 and 0·8 mM, respectively.

Article Snippet: HT-29 human colon adenocarcinoma cells (American Type Culture Collection (ATCC), Rockville, MD, USA) were grown in minimum essential medium.

Techniques: Comparison

Journal: British Journal of Nutrition

Article Title: Effect of β-carotene-rich tomato lycopene β-cyclase (tlcy-b) on cell growth inhibition in HT-29 colon adenocarcinoma cells

doi: 10.1017/s0007114508169902

Figure Lengend Snippet: Fig. 4. Effect of b-carotene on cell growth (A), caspase-3 activation (B), Bcl-2 (C) and Bcl-xl (D) content in HT-29 cells following a 24 h treatment. Values are the means of three different experiments, with standard errors represented by vertical bars. a,b,c Mean values with unlike letters were significantly different (P,0·05; Fischer’s test).

Article Snippet: HT-29 human colon adenocarcinoma cells (American Type Culture Collection (ATCC), Rockville, MD, USA) were grown in minimum essential medium.

Techniques: Activation Assay

Journal: Nature Communications

Article Title: Search-and-remove genome editing allows selection of cells by DNA sequence

doi: 10.1038/s41467-025-66896-1

Figure Lengend Snippet: a Strategy to apply SNIPE to kill cancer cells based on cancer specific mutations. Wild type cells are colored in gray, cells with cancer mutations in purple and dying cells in red. b Cell growth curves of human 409B2 iPSCs measured by resazurin assay (upper panel). The lower panel shows NGS read frequency for cancer mutation (purple) and wild type cells (gray) in a mixed population over time. c Killing of TP53 knock-out 409B2 cells by SNIPE. Different amounts of TP53 knock-out cells (purple) were mixed with wild type cells (gray). Indels are colored in blue and wild type with indels in light purple. d Cell survival after selection by SNIPE related to ( c ). e K562 cells carry a Philadelphia chromosome, characterized by a reciprocal translocation of chromosome 9 and 22 resulting in the fusion oncogene BCR-ABL1 . f The BCR-ABL fusion generates a new gRNA recognition site in K562 cells. g Cell survival of THP-1 and K562 cells after targeting the BCR-ABL junction. The label “Cas9 cut” indicates CRISPR targeting without SNIPE. h Agarose gel of PCR products of healthy chromosome 22 (297 bp, gray) and Philadelphia chromosome (267 bp, purple). i Quantification of band intensity for K562 cells of ( h ). j Cell survival of heterozygous cancer cell lines RD-ES, HT-29 and HuCC-T1 after targeting the disease allele with SNIPE. k NGS read frequency of cancer locus alleles in RD-ES, HT-29 and HuCC-T1 for mock and SNIPE edit. The cancer mutation allele is colored in purple, wild type in grey, indels in blue and wild type with indels in light purple. Independent biological replicates were performed ( n = 3 for b , c , d , g , h , i , j , k , n = 2 for RDES in j and k) and are depicted as dots for the cancer mutation allele sequence reads and cell survival. Error bars show the s.e.m.

Article Snippet: Human colorectal adenocarcinoma cells HT-29 (Cytion, catalog no. 300215) were cultured in EMEM (Sigma-Aldrich, catalog no. M4655) supplemented with 10% FBS (Gibco, catalog no. 10270-106) and human CML cells K562 (ECACC, catalog no. 89121407) were cultured in IMDM (Gibco, catalog no. 12440053) with 10% FBS.

Techniques: Resazurin Assay, Mutagenesis, Knock-Out, Selection, Translocation Assay, CRISPR, Agarose Gel Electrophoresis, Sequencing

Journal: Theranostics

Article Title: Proteomic analysis of intracellular protein corona of nanoparticles elucidates nano-trafficking network and nano-bio interactions

doi: 10.7150/thno.38900

Figure Lengend Snippet: Nano-trafficking mechanism of AuNPs verified by conventional approaches. (A) Analysis of the endocytosis mechanism of AuNPs following treatment with different chemical inhibitors. Mean ± SD, n = 3, * p < 0.05 and ** p < 0.01. (B) Colocalization images of AuNPs with various organelles (lysosome, endoplasmic reticulum, Golgi apparatus, mitochondrion). Scale bar CLSM, 10 μm. (C) Colocalization coefficients of AuNPs with various organelles (lysosome, endoplasmic reticulum, Golgi apparatus, mitochondrion) indicating percent AuNPs which colocalized with organelles. Mean ± SD, n = 8. (D) Cellular distribution of AuNPs under TEM (left) and its schematic diagram (right). Arrows indicate the AuNPs. Scale bar TEM, 500 nm. (E) Colocalization images of AuNPs with specific IPC-Trans proteins (CLTB, FN1, IGFBP4, and PSAP). Scale bar CLSM, 10 μm. (F) Colocalization coefficients of AuNPs with different IPC-Trans proteins. The colocalization coefficient indicates percent AuNPs which colocalized with IPC-Trans proteins. Mean ± SD, n = 8.

Article Snippet: CLTB mouse monoclonal antibody (66270-1-Ig, 1:50), IGFBP4 rabbit polyclonal antibody (18500-1-AP, 1:20), and PSAP rabbit polyclonal antibody (10801-1-AP, 1:50) were purchased from Proteintech (Chicago, USA).

Techniques: