prima-1 met  (SAS institute)

Journal: bioRxiv

Article Title: Dysregulation of the p53 pathway provides a therapeutic target in aggressive pediatric sarcomas with stem-like traits

doi: 10.1101/2024.01.17.576012

Figure Lengend Snippet: a) Representative images of western blot analysis (biological n=3) of stemness-associated TFs SOX2, NANOG, OCT4, KLF4 and c-MYC and p53 pathway proteins, p53 and MDM2, in a panel of established and patient-derived OS, RMS and ES cell lines. Densitometric analysis is provided in Fig. S1. b) Heat maps (top) of the mean protein expression (n=3) of stemness-related TFs, p53 and MDM2. Cell lines of each sarcoma subgroup are aligned according to the calculated SI (middle). The scheme (bottom) illustrates the tumorigenicity of SI high and SI low cell lines assayed in NSG mice (n=3). c) Survival of xenograft mouse models (n=3 mice per cell line) reflecting tumor-initiating capacity and xenograft growth rates of the sarcoma cell lines tested. d) A significant positive correlation between the expression of SOX2, OCT4 and NANOG TFs and number of tumors formed by sarcoma cell lines in mice. e) Correlation between SI values and p53 protein level in individual cell lines relative to the mean expression of each sarcoma subtype. Note that all tumorigenic SI high models exhibit deregulated p53 expression and fit within the upper or lower quartiles (indicated by dotted lines). f) p53 levels in SI high models are significantly different compared to the rest of the sarcoma cell lines. Data are presented as mean ± SD. Statistical significance was determined by one-tailed Welch’s t-test (f) and by Spearman correlation coefficient (d) , *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Drugs targeting p53 pathway were used for the treatment: RO-5963 (MDM2/MDMX dual inhibitor, #444153, Sigma-Aldrich, St. Louis, MO, USA) and PRIMA-1 MET (mut-p53 reactivator, #SML1789, Sigma-Aldrich).

Techniques: Western Blot, Derivative Assay, Expressing, One-tailed Test

Journal: bioRxiv

Article Title: Dysregulation of the p53 pathway provides a therapeutic target in aggressive pediatric sarcomas with stem-like traits

doi: 10.1101/2024.01.17.576012

Figure Lengend Snippet: a) An overview of the in-house pipeline for TMA assembly followed by IHC staining and machine learning-assisted image analysis of the whole-slide scanned TMA sections. b, c) Staining intensity of p53 and stemness-related TFs assessed by computational analysis in QuPath software ( b ) or manual readout by experienced pathologist ( c ). Data shown as mean ± SD, biological n=3. d) Representative images of IHC detection of SOX2, KLF4 and p53 in xenograft tumors formed by the indicated cell lines or in positive control tissues (SOX2, fetal lungs; KLF4, seminoma; p53, tonsilla).

Article Snippet: Drugs targeting p53 pathway were used for the treatment: RO-5963 (MDM2/MDMX dual inhibitor, #444153, Sigma-Aldrich, St. Louis, MO, USA) and PRIMA-1 MET (mut-p53 reactivator, #SML1789, Sigma-Aldrich).

Techniques: Immunohistochemistry, Staining, Software, Positive Control

Journal: bioRxiv

Article Title: Dysregulation of the p53 pathway provides a therapeutic target in aggressive pediatric sarcomas with stem-like traits

doi: 10.1101/2024.01.17.576012

Figure Lengend Snippet: a) Representative western blot images (right) and densitometric analysis (left) of stemness-associated TFs, p53 and MDM2 in mut-p53 MNNG/HOS and wt-p53 ESFT-15 clones with shRNA-mediated knockdown of SOX2 (shSOX2) compared with their scramble shRNA controls (shCtrl). Data presented as mean ± SD, biological n=3–4. Densitometric analysis of OCT4, c-MYC, and MDM2 proteins is provided in Fig. S10a. b) qPCR analysis of selected stemness-associated TFs in MNNG/HOS and ESFT-15 shSOX2 clones. Data are presented as mean ± SD, biological n=3, technical n=3. Analysis of remaining genes is provided in Fig. S10b. c) Sphere formation assay evaluating stem-like phenotype of MNNG/HOS and ESFT-15 sarcoma cells after SOX2 knockdown. Stem cell frequencies and probability were computed using ELDA software . Data are shown as mean ± 95% confidence interval, biological n=3–4, technical n=5. d) Representative images of sarcospheres formed by shCtrl and shSOX2 clones of MNNG/HOS and ESFT-15 cell lines. All experiments were performed using single cell-derived clones (indicated by numbers). Statistical significance was determined by one-way ANOVA with Welch’s correction followed by post-hoc Dunnett’s test ( a, b ) or Chi-square pairwise test ( c ), *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Drugs targeting p53 pathway were used for the treatment: RO-5963 (MDM2/MDMX dual inhibitor, #444153, Sigma-Aldrich, St. Louis, MO, USA) and PRIMA-1 MET (mut-p53 reactivator, #SML1789, Sigma-Aldrich).

Techniques: Western Blot, Clone Assay, shRNA, Tube Formation Assay, Software, Derivative Assay

Journal: bioRxiv

Article Title: Dysregulation of the p53 pathway provides a therapeutic target in aggressive pediatric sarcomas with stem-like traits

doi: 10.1101/2024.01.17.576012

Figure Lengend Snippet: a) MTT cell viability assay of SI high and SI low OS, RMS and ES cell lines after 72-h treatment by RO-5963 or PRIMA-1 MET . Note that only cells derived from ES showed marked difference in sensitivity to re-activation of p53. Data are presented as mean ± SD, biological n=3–5, technical n=3–5. b) MTT cell viability assay of control healthy fibroblast cell lines after 72-h exposure to RO-5963 or PRIMA-1 MET . Data are presented as mean ± SD, biological n=3, technical n=3–5. c, d) Representative western blot images ( c ) and densitometric analysis ( d ) of stemness-associated TFs expression in aggressive ESFT-15 cells after 72-h treatment with RO-5963 or PRIMA-1 MET . Ctrl, DMSO treated control. Data shown as mean ± SD, biological n=3. Statistical significance was determined by one-way ANOVA with Welch’s correction followed by post-hoc Dunnett’s test, **p<0.01.

Article Snippet: Drugs targeting p53 pathway were used for the treatment: RO-5963 (MDM2/MDMX dual inhibitor, #444153, Sigma-Aldrich, St. Louis, MO, USA) and PRIMA-1 MET (mut-p53 reactivator, #SML1789, Sigma-Aldrich).

Techniques: Viability Assay, Derivative Assay, Activation Assay, Western Blot, Expressing

Journal: bioRxiv

Article Title: Dysregulation of the p53 pathway provides a therapeutic target in aggressive pediatric sarcomas with stem-like traits

doi: 10.1101/2024.01.17.576012

Figure Lengend Snippet: a, b) Representative western blot images (a) and densitometric analysis (b) of p53, MDM2 and cleaved caspase-3 expression in ESFT-15 ES cell line and healthy fibroblasts NDF-3 and NDF-2 after 72-h exposure to RO-5963 and PRIMA-1 MET . Data are presented as mean ± SD, biological n=3. c) Representative flow cytometry histograms showing marked difference in SYTOX Red-positive dead cells in ESFT-15 and fibroblast cell lines after 72-h exposure to 10×IC 50 (ESFT-15) of RO-5963. d) Flow cytometry analysis of dead vs. live cells after 72-h treatment with p53 pathway targeting drugs, RO-5963 and PRIMA-1 MET . Data presented as mean ± SD, biological n=3-6. Ctrl, DMSO treated control. Statistical significance in was determined by one-way ANOVA with Welch’s correction followed by post-hoc Dunnett’s test (b,d), *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Drugs targeting p53 pathway were used for the treatment: RO-5963 (MDM2/MDMX dual inhibitor, #444153, Sigma-Aldrich, St. Louis, MO, USA) and PRIMA-1 MET (mut-p53 reactivator, #SML1789, Sigma-Aldrich).

Techniques: Western Blot, Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Dysregulation of the p53 pathway provides a therapeutic target in aggressive pediatric sarcomas with stem-like traits

doi: 10.1101/2024.01.17.576012

Figure Lengend Snippet: a) IHC analysis of SOX2 and KLF4 expression in OS, RMS and ES biopsies. b) Expression of KLF4 and SOX2 in p53-positive and p53-negative OS and ES tissues, respectively. Note that in ES, SOX2 was detected only in p53-positive biopsies. c) Representative images of SOX2 and p53 IHC analysis in ES tissue. d) Frequency of initial metastases in OS patients with KLF4- positive and KLF4-negative tumors. e) Kaplan-Meier analysis of OS patients stratified by KLF4 and p53 protein expression. f) Frequency of initial metastases in ES patients with SOX2 high and SOX2 low tumors. g) Kaplan-Meier analysis of ES patients stratified by SOX2 protein expression (cut-off for SOX2 low histoscore value was determined as ≤ 45). h) Frequency of initial metastases in pediatric sarcoma patients with p53-positive and p53-negative tumors, showing that p53 expression is associated with early metastatic disease. i) Kaplan-Meier analysis of sarcoma patients stratified by p53 protein expression and presence of initial metastases. M0, no metastatic events; M1, presence of initial metastases. Statistical significance was determined by Welch’s t-test ( a, b ), Chi-square test ( d, f, h ) and Mantel-Cox test ( e, g, i ), *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Drugs targeting p53 pathway were used for the treatment: RO-5963 (MDM2/MDMX dual inhibitor, #444153, Sigma-Aldrich, St. Louis, MO, USA) and PRIMA-1 MET (mut-p53 reactivator, #SML1789, Sigma-Aldrich).

Techniques: Expressing

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: Selection of PRIMA-1 MET and PTC596 dose treatments. Cell viability was evaluated using MTS assay in cells exposed to increasing concentrations of the drugs. ( a ) HTLA-230 and HTLA-ER cells were treated with 10–60 µM of PRIMA-1 MET for 48 h and 72 h. ( b ) HTLA-ER cells were treated with 20–200 nM of PTC596 for 48 h and 72 h. Results are expressed as percentage variations in cell viability of treated cells with respect to that of untreated ones (100%). Graphs summarize quantitative data of means ± SEM of at least four independent experiments. **** p < 0.0001 vs. untreated HTLA-230 or HTLA-ER cells.

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: Selection, MTS Assay

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: PRIMA-1 MET and PTC596, used in combination, were cytotoxic to HTLA-230 and HTLA-ER cells. Cell viability was evaluated using MTS assay in HTLA-230 ( a ) and HTLA-ER ( b ) cells treated with 1.25 μM of etoposide (24 h), 60 μM of PRIMA-1 MET (48 h and 72 h), and 35 nM of PTC596 (48 h and 72 h), given alone or in each possible combination. Results are expressed as percentage variations in cell viability of treated cells with respect to that of untreated ones (100%). Histograms summarize quantitative data of means ± SEM of at least four independent experiments. * p < 0.1; **** p < 0.0001 vs. untreated HTLA-230 or HTLA-ER cells (Ctr); °°° p < 0.001; °°°° p < 0.0001 vs. etoposide treated cells; # p < 0.0001 vs. respective treatments indicated by the bars.

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: MTS Assay

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: PTC596, alone or in combination, reduced the expression levels of BMI-1, while PRIMA-1 MET did not alter the expression of P53 and its related proteins. Protein levels of BMI-1 ( a ), P53 ( b ), p21, and MDM2 ( c ) in HTLA-230 (left panel) and HTLA-ER (right panel) cells untreated and treated with 1.25 μM of etoposide (24 h), 60 μM of PRIMA-1 MET (48 h), and 35 nM of PTC596 (72 h), given alone or in combination. Immunoblots shown are representative of four independent experiments. GAPDH ( a ) or tubulin ( b , c ) were the internal loading controls used to normalize the protein levels of BMI-1 and P53, respectively. Results are expressed as percentage variations in protein levels in treated cells with respect to that in untreated ones (100%). Histograms summarize quantitative data of protein level means, normalised to the respective loading control ± SEM of four independent experiments. ** p < 0.01; *** p < 0.001 vs. untreated cells (Ctr); ° p < 0.0001 vs. etoposide-treated cells.

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: Expressing, Western Blot

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: PTC596, alone or in combination, does not induce apoptosis. Protein levels of Bax and Bcl-2 ( a ) in HTLA-230 (left panel) and HTLA-ER (right panel) cells untreated or treated with 1.25 μM of etoposide (24 h), 60 μM of PRIMA-1 MET (48 h), and 35 nm of PTC596 (72 h), given alone or in combination. The histogram reported in ( b ) summarizes the values of Bax/Bcl-2 ratio. Immunoblots shown are representative of four independent experiments. Tubulin is the internal loading control. Results are expressed as percentage variations in protein levels in treated cells with respect to that in untreated ones (100%). Histograms summarize quantitative data of protein level means, normalised to tubulin ± SEM of four independent experiments. * p < 0.1; ** p < 0.01; *** p < 0.001 vs. untreated cells (Ctr).

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: Western Blot

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: PRIMA-1 MET and PTC596, alone or in combination, inhibit the clonogenic potential of HTLA-230 and HTLA-ER cells. The ability to form colonies was analysed by using an anchorage-independent clonogenic assay. HTLA-230 ( a ) and HTLA-ER ( b ) cells were treated with 1.25 μM of etoposide (24 h), 60 μM of PRIMA-1 MET (48 h), and 35 nM of PTC596 (72 h), given alone or in combination. Results are expressed as percentage variations in colony number of treated cells with respect to that of untreated ones (100%). The histograms summarize quantitative data of means ± SEM of four independent experiments. **** p < 0.0001 vs. untreated cells (Ctr); # p < 0.1; #### p < 0.0001 vs. respective treatments indicated by the bars.

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: Clonogenic Assay

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: PRIMA-1 MET and PTC596, alone or in combination, inhibit the expression of EMT-related proteins. Protein levels of N-cadherin ( a ), ß-catenin ( b ), and SNAIL ( c ) in HTLA-230 (left panels) and HTLA-ER (right panels) cells untreated or treated with 1.25 μM of etoposide (24 h), 60 μM of PRIMA-1 MET (48 h), and 35 nM of PTC596 (72 h), given alone or in combination. Immunoblots shown are representative of four independent experiments. Tubulin is the internal loading control. Results are expressed as percentage variations in protein levels in treated cells with respect to that in untreated ones (100%). The histograms summarize quantitative data of protein level means, normalised to tubulin ± SEM of four independent experiments. * p < 0.1; ** p < 0.01; *** p < 0.001; **** p < 0.0001 vs. untreated cells (Ctr); °° p < 0.01; °°° p < 0.001 vs. etoposide-treated cells; # p < 0.1; ### p < 0.001; #### p < 0.0001 vs. PRIMA-1 MET -treated cells.

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: Expressing, Western Blot

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: PRIMA-1 MET and PTC596, alone or in combination, totally counteract CSC generation. Evaluation of CSC number in HTLA-230 and HTLA-ER cells untreated or treated with 1.25 μM of etoposide (24 h), 60 μM of PRIMA-1 MET (48 h), and 35 nM of PTC596 (72 h), given alone or in combination. The number of cells obtained by the disaggregation of CSCs, generated after 4 weeks, is reported as percentage values of the number of treated cells in comparison with those of untreated ones (100%). The histograms summarize quantitative data of means ± SEM of four independent experiments, 4 weeks after the treatments. *** p < 0.001; **** p < 0.0001 vs. untreated cells (Ctr).

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: Generated, Comparison

Journal: Antioxidants

Article Title: PTC596-Induced BMI-1 Inhibition Fights Neuroblastoma Multidrug Resistance by Inducing Ferroptosis

doi: 10.3390/antiox13010003

Figure Lengend Snippet: PRIMA-1 MET and PTC596, alone or in combination, enhance H 2 O 2 production, reduce GSH intracellular levels, and induce lipoperoxidation. H 2 O 2 production ( a ), GSH levels ( b ), and lipid peroxidation (BODIPY-positive cells ( c ) were analysed in HTLA-230 and HTLA-ER cells untreated or treated with 1.25 μM of etoposide (24 h), 60 μM of PRIMA-1 MET (48 h), and 35 nM of PTC596 (72 h), given alone or in combination). Results are expressed as percentage variations in DCFH-positive cells, GSH levels, or BODIPY-positive cells under treatment conditions in comparison with untreated ones (100%). Histograms summarise quantitative data of means ± SEM of four independent experiments. * p < 0.1; ** p < 0.01; *** p < 0.001; **** p < 0.0001 vs. untreated cells (Ctr); ° p < 0.1; °° p < 0.01; °°°° p < 0.0001 vs. etoposide-treated cells; #### p < 0.0001 vs. PRIMA-1 MET -treated cells; ^^^^ p < 0.0001 vs. PTC596-treated cells.

Article Snippet: Etoposide was purchased from Calbiochem (Merk KGaA, Darmstadt, Germany), PRIMA-1 MET from Abcam (Cambridge, UK), and PTC596 from PTC Therapeutics (South Plainfield, NJ, USA).

Techniques: Comparison

Journal: Life Science Alliance

Article Title: APR-246 increases tumor antigenicity independent of p53

doi: 10.26508/lsa.202301999

Figure Lengend Snippet: (A) The schematic for the vaccination of C57BL/6 mice with B16-GMCSF cells that had undergone an in vitro treatment with MQ and adoptive transfer of Pmel CD8 + T cells. (A, B) Left panel—representative flow images for CFSE dilution of pmel CD8 + T cells harvested from the draining lymph node of C57BL/6 mice implanted with B16-GMCSF cells according to the schematic in (A). (B) Middle panel—fraction of CFSE low (proliferating) pmel CD8 + T cells among total CFSE + pmel CD8 + T cells. (B) Right panel—proportion of pmel CD8 + T cells among total CD8 + cells in the lymph node. (C) Mean fluorescence intensity (MFI) (MFI of antibody/MFI of isotype control) of the MHC class I (H2Kb) and the MHC class II (IA/IE) as evaluated by flow cytometry in (C). (D) B16 cells that were treated in vitro with MQ (10 μM)/vehicle for 4 h and harvested 24 h later or (D) in B16 tumors from mice treated with APR-246 (pulse treatment of 100 mg/kg 2qd biweekly x2). (E) Effect of in vitro MQ treatment on the Trp53 pathway in B16-derived WT and Trp53 CRISPR/Cas9 knock-out (Trp53 −/− ) cells as measured by Western blot. The treatment was done either in a continuous or pulse fashion (pulse treatment with MQ/vehicle for 2–4 h, drug removal, and cell culture in fresh media for 24/48 h). CDDP, cisplatin (40 μM) treatment for 24 h was used as a positive control for p53 activation. β-Actin was used as a loading control. (F) The MFI of the MHC class I (H2Kb) and the MHC class II (IA/IE) as evaluated by flow cytometry in B16 WT or Trp53 −/− cells when treated in vitro with MQ (10 μM)/vehicle for 4 h and harvested 24 h later. The data represent mean ± SEM and the P -value was calculated by unpaired parametric t test for the in vivo data and two-way ANOVA test for the in vitro data, *<0.0332, **<0.0021, ***<0.0002, ****<0.0001. n = 3 for in vitro experiments or n = 5 for in vivo experiments and is representative of three independent experiments.

Article Snippet: APR-246/PRIMA-1 MET (eprenetapopt, Aprea Therapeutics) is a novel small molecule anti-cancerous compound that stabilizes p53 in the active conformation by conjugation to thiol groups (AKA sulfhydryl [SH] groups) ( ; ).

Techniques: In Vitro, Adoptive Transfer Assay, Fluorescence, Flow Cytometry, Derivative Assay, CRISPR, Knock-Out, Western Blot, Cell Culture, Positive Control, Activation Assay, In Vivo

Journal: Life Science Alliance

Article Title: APR-246 increases tumor antigenicity independent of p53

doi: 10.26508/lsa.202301999

Figure Lengend Snippet: (A, B) The schematic for the combination of the intratumoral injection (i.t.) of the TLR4 agonist, monophosphoryl lipid A (MPLA) with either APR-246 intra-peritoneal (i.p.) continuous administration (one a day 1qd, every day) or (B) the APR-246 pulse administration (twice a day 2qd biweekly). (C) The corresponding tumor growth curves (left panel) and survival of mice (right panel) for the respective treatment combinations. Triethylamine i.t. injection is used as a vehicle control for MPLA. (D) The phenotype of the tumor infiltrating myeloid and lymphoid cells under aforementioned treatment schedule as assessed using flow cytometry in the MPLA-injected tumors. (E, F) B16 WT and (F). Trp53 −/− B16 non-injected tumor growth curves for the triple combination of i.t. MPLA and CD40 agonist (clone FGK45, BioXcell) (CD40a) with i.p. pulse APR-246. An isotype was used as negative control for CD40 agonist (clone 2A3, BioXcell). The data represent mean ± SEM and the P -value is represented as *<0.0332, **<0.0021, ***<0.0002, ****<0.0001. P -value was calculated by two-way ANOVA, by log-rank (Mantel–Cox) test for survival curves. (C, D) Representative experiment of three independent experiments, n = 10 for (C) and n = 5 for (D).

Article Snippet: APR-246/PRIMA-1 MET (eprenetapopt, Aprea Therapeutics) is a novel small molecule anti-cancerous compound that stabilizes p53 in the active conformation by conjugation to thiol groups (AKA sulfhydryl [SH] groups) ( ; ).

Techniques: Injection, Flow Cytometry, Negative Control

Journal: Life Science Alliance

Article Title: APR-246 increases tumor antigenicity independent of p53

doi: 10.26508/lsa.202301999

Figure Lengend Snippet: (A) Schematic representation of the corresponding treatment schedule for the intra-peritoneal (i.p.) administration of APR-246 and the intratumoral injection (i.t.) of the TLR4 agonist, monophosphoryl lipid A (MPLA) in a bilateral flank tumor model. (B, C) The mean tumor size (B) and overall survival of mice (C) under the aforementioned combination treatment. Triethylamine i.t. injection was used as a vehicle control for MPLA. (D) The phenotype of the tumor-infiltrating myeloid and lymphoid cells under the aforementioned treatment schedule was assessed in the MPLA-injected tumors using flow cytometry. (E) Treatment schematic for the triple combination of i.t. MPLA and CD40 agonist (clone FGK45, BioXcell) with i.p. pulse APR-246. An isotype was used as negative control for CD40 agonist (clone 2A3, BioXcell). (F, G) The corresponding tumor growth and mice survival analysis of (F) WT and (G) Trp53 −/− B16 tumors. The data represent mean ± SEM and the P -value is represented as *<0.0332, **<0.0021, ***<0.0002, ****<0.0001. P -value was calculated by two-way ANOVA, by log-rank (Mantel–Cox) test for survival curves. n = 10 for (B, E) and n = 5 for (D) and representative of two (D, E) to three (B) independent experiments.

Article Snippet: APR-246/PRIMA-1 MET (eprenetapopt, Aprea Therapeutics) is a novel small molecule anti-cancerous compound that stabilizes p53 in the active conformation by conjugation to thiol groups (AKA sulfhydryl [SH] groups) ( ; ).

Techniques: Injection, Flow Cytometry, Negative Control