bt474 breast cancer cell lines  (ATCC)

Journal: Clinical Cancer Research

Article Title: Ultrasensitive Detection and Monitoring of Circulating Tumor DNA Using Structural Variants in Early-Stage Breast Cancer

doi: 10.1158/1078-0432.CCR-24-3472

Figure Lengend Snippet: Contrived cfDNA samples prepared for assessment of assay analytical validity. A, dPCR output of the LoD95 study at a dilution of 0.0005% (5 ppm) in BT474 demonstrating 100% ctDNA detection. Illustration of replicate number (1–32) vs. SV number (1–16). Dark green–filled cells indicate positive SV results. B, Example dPCR data (1D plots) for one positive SV result, including positive and negative controls, are shown. A representative threshold value is illustrated.

Article Snippet: A fingerprint was designed for the breast cancer cell line BT474 (HER2 + breast carcinoma, Cytion—Cell Lines Service GmbH, product ID: 300131GD5; RRID: CVCL_0179), and LoD95 was determined using the probit method for a standard cfDNA input amount (70 ng, RRID: CVCL_1C78, product ID: NA24385, Coriell DNA, fragmented to mimic cfDNA).

Techniques:

Journal: Clinical Cancer Research

Article Title: Ultrasensitive Detection and Monitoring of Circulating Tumor DNA Using Structural Variants in Early-Stage Breast Cancer

doi: 10.1158/1078-0432.CCR-24-3472

Figure Lengend Snippet: Baseline participant characteristics in the EBC clinical validation cohort.

Article Snippet: A fingerprint was designed for the breast cancer cell line BT474 (HER2 + breast carcinoma, Cytion—Cell Lines Service GmbH, product ID: 300131GD5; RRID: CVCL_0179), and LoD95 was determined using the probit method for a standard cfDNA input amount (70 ng, RRID: CVCL_1C78, product ID: NA24385, Coriell DNA, fragmented to mimic cfDNA).

Techniques: Diagnostic Assay

Journal: Clinical Cancer Research

Article Title: Ultrasensitive Detection and Monitoring of Circulating Tumor DNA Using Structural Variants in Early-Stage Breast Cancer

doi: 10.1158/1078-0432.CCR-24-3472

Figure Lengend Snippet: Swimmer plots and clinical events. Participants’ clinical timeline and timeline of plasma collection for ctDNA analysis for ( A ) ER + , ( B ) TNBC, and ( C ) HER2 + EBC. Plots are broken down by participants with and without clinical recurrence and individual stage (I, II, and III) as well as RCB is defined. FUP, follow-up.

Article Snippet: A fingerprint was designed for the breast cancer cell line BT474 (HER2 + breast carcinoma, Cytion—Cell Lines Service GmbH, product ID: 300131GD5; RRID: CVCL_0179), and LoD95 was determined using the probit method for a standard cfDNA input amount (70 ng, RRID: CVCL_1C78, product ID: NA24385, Coriell DNA, fragmented to mimic cfDNA).

Techniques:

Journal: Journal of Nuclear Medicine

Article Title: Preirradiation of Spheroids with 225 Ac-Trastuzumab Improves Penetration of 225 Ac-Liposomes and MIRDcell Predictions of Responses to Drug Cocktails

doi: 10.2967/jnumed.124.269273

Figure Lengend Snippet: Penetration data in BT474 spheroids (200-µm radius) for Alexa Fluor 647-NHS-trastuzumab ( ), CFDA-SE liposomes ( ), and CFDA-SE liposomes after preirradiating spheroids with 6.5 kBq/mL 225 Ac-trastuzumab ( ). Shown are time-integrated average concentrations as function of distance from centers of spheroids (A) and correlated mean decays per cell using activity concentrations of 6.875 kBq/mL for all drugs (calculated with MIRDcell-Ã) (B). Distances beyond 200 µm represent medium.

Article Snippet: The spheroids comprised the human epidermal growth factor receptor 2–positive human BT474 breast cancer cell line, obtained from the American Type Culture Collection.

Techniques: Liposomes, Activity Assay

Journal: Journal of Nuclear Medicine

Article Title: Preirradiation of Spheroids with 225 Ac-Trastuzumab Improves Penetration of 225 Ac-Liposomes and MIRDcell Predictions of Responses to Drug Cocktails

doi: 10.2967/jnumed.124.269273

Figure Lengend Snippet: MIRDcell calculated mean decays per cell attributed to liposomes in BT474 spheroids based on experimental CFDA-SE liposome penetration data for (100% 225 Ac-liposome case from Howe et al. ) and (50% 225 Ac-liposome, 50% 225 Ac-trastuzumab case from newly retrieved data), and extrapolated/averaged (weighted) liposome penetration data for (70% 225 Ac-liposome, 30% 225 Ac-trastuzumab case) and (30% 225 Ac-liposome, 70% 225 Ac-trastuzumab case).

Article Snippet: The spheroids comprised the human epidermal growth factor receptor 2–positive human BT474 breast cancer cell line, obtained from the American Type Culture Collection.

Techniques: Liposomes

Journal: Journal of Nuclear Medicine

Article Title: Preirradiation of Spheroids with 225 Ac-Trastuzumab Improves Penetration of 225 Ac-Liposomes and MIRDcell Predictions of Responses to Drug Cocktails

doi: 10.2967/jnumed.124.269273

Figure Lengend Snippet: Comparison of BT474 experimental outgrowths taken from Howe et al. to MIRDcell (version 4.14)-predicted SFs as bar plot (A), linear regression plot (B), and Bland–Altman plot (C) (95% limit of agreement, −0.048 to 0.095). Predicted SFs shown were based on 225 Ac-trastuzumab in spheroids with no 225 Ac daughters present and 225 Ac-liposomes in spheroids with 225 Ac daughters present. 225 Ac-trastuzumab and 225 Ac-liposomes in medium both had 225 Ac daughters present. Bars on abscissa of panel A and legend for panels B and C are labeled to denote percentage of activity corresponding to 225 Ac-liposomes and 225 Ac-trastuzumab. For example, “L-100, A-0” is 100% 225 Ac-liposomes and 0% activity on trastuzumab antibodies. Error bars for experimental outgrowths correspond to SD. Final prediction uses penetration data for CFDA-SE liposomes after spheroids are exposed to 6.5 kBq/mL 225 Ac-trastuzumab for 50%/50% case; 70% and 30% 225 Ac-liposome cases use extrapolated/averaged penetration data shown in . For CFDA-SE liposomes, published penetration data that were used for old prediction did not involve pretreatment 225 Ac-trastuzumab. Only final predictions are shown in panels B and C.

Article Snippet: The spheroids comprised the human epidermal growth factor receptor 2–positive human BT474 breast cancer cell line, obtained from the American Type Culture Collection.

Techniques: Comparison, Liposomes, Labeling, Activity Assay

Journal: Journal of Nuclear Medicine

Article Title: Preirradiation of Spheroids with 225 Ac-Trastuzumab Improves Penetration of 225 Ac-Liposomes and MIRDcell Predictions of Responses to Drug Cocktails

doi: 10.2967/jnumed.124.269273

Figure Lengend Snippet: Mean absorbed dose due to 225 Ac-trastuzumab (antibodies) and 225 Ac-liposomes (liposomes) in BT474 spheroids and medium for cells at various radial depths within 200-µm-radius spheroids (top). Below each graph, corresponding MIRDcell 3D slice representation of spheroid is shown for equatorial slices of spheroids. Dark red dots in equatorial slices represent living cells, whereas pink dots represent dead cells.

Article Snippet: The spheroids comprised the human epidermal growth factor receptor 2–positive human BT474 breast cancer cell line, obtained from the American Type Culture Collection.

Techniques: Liposomes

Journal: Journal of Nuclear Medicine

Article Title: Preirradiation of Spheroids with 225 Ac-Trastuzumab Improves Penetration of 225 Ac-Liposomes and MIRDcell Predictions of Responses to Drug Cocktails

doi: 10.2967/jnumed.124.269273

Figure Lengend Snippet: MIRDcell (version 4.16) AI–predicted minimum number of decays necessary to achieve target SF < 0.0001 in BT474 spheroids treated with 225 Ac-trastuzumab (antibodies), 225 Ac-liposomes (liposomes), or combination thereof. 225 Ac daughters were assumed to migrate away from 225 Ac-trastuzumab and to remain near 225 Ac-liposomes within spheroid. MIRDcell AI was limited to maximum molar activity of 10 6 GBq/mol for both drugs and lower limit of 1.17 × 10 5 GBq/mol for antibodies only. No bar appearing for antibodies indicates that 225 Ac-trastuzumab alone could not achieve target SF. When optimized molar activities for combined therapy and liposome-only therapy were checked in Monte Carlo–based MIRDcell simulation, zero cell survivors were achieved for both therapies although liposomes required more total decays to achieve same result. “Liposome with preirradiation” indicates use of penetration profiles after preirradiation with 225 Ac-trastuzumab as in 50%/50% case, and “Liposome without preirradiation” indicates use of liposome penetration profile without preirradiation as in 100% 225 Ac-liposomes case.

Article Snippet: The spheroids comprised the human epidermal growth factor receptor 2–positive human BT474 breast cancer cell line, obtained from the American Type Culture Collection.

Techniques: Liposomes, Activity Assay

Journal: Cancer Immunology Research

Article Title: Antitumor CD4 + T Helper 1 Cells Target and Control the Outgrowth of Disseminated Cancer Cells

doi: 10.1158/2326-6066.CIR-24-0630

Figure Lengend Snippet: Gene expression status of DCCs, primary tumors, and metastatic tumors. A, Immunofluorescence staining for HER2 + CYT8/18 + Ki-67 + DCCs detection in the BM. Scale bars, 100 μm. B, Schematic showing the methods for DCCs isolation from the BM of BALB-neuT mice. C, Percentage of EpCAM + DCCs isolated from the BM of BALB-neuT mice as determined by flow cytometry (isolated from the BM pool of 10 BALB-neuT mice). D, Cumulative representation of differentially expressed cancer stemness, EMT, and cell-cycle genes in DCCs of BALB-neuT mice, early lesion tumor cells from BALB-neuT mice (EL), and metastatic TUBO cells analyzed by RNA sequencing ( n = 2/group). Cumulative Z -score denotes average score of genes in the respective panels. E, Schematic depicting the methods for DCCs isolation from the BM aspirates of patients with HER2 + breast cancer (BC). F, Cytopathologic confirmation of isolated DCCs from the BM of patients with HER2 + BC by Diff-Quik staining. Scale bars, 100 μm. G, Immunofluorescence staining for HER2 + CYT8/18 + DCCs detection in the BM of patients with HER2 + BC. Scale bars, 100 μm. H, Percentage of Pan-cytokeratin + (Pan-CYT + ) DCCs isolated from the BM of a patient with HER2 + BC analyzed by flow cytometry. I, Bright-field image of in vitro cultured DCCs of patients with BC. Scale bars, 500 μm. J, Cumulative representation (average Z -score of genes) of differentially expressed cancer stemness, EMT, and cell-cycle genes in DCCs of patients with HER2 + BC, JIMT-1, BT474, and HCC1954 cells analyzed by RNA sequencing ( n = 3/group). All data are presented as mean ± SEM. P values were assessed by one-way ANOVA with Tukey multiple comparisons test. SSC-A, side scatter area.

Article Snippet: The human breast cancer cell lines BT474 (Cat. No. HTB20) and HCC1954 (Cat. No. CRL-2338) were purchased from ATCC in 2018.

Techniques: Gene Expression, Immunofluorescence, Staining, Isolation, Flow Cytometry, RNA Sequencing, Diff-Quik, In Vitro, Cell Culture