Journal: Molecular Therapy Oncology

Article Title: Strategically engineering an oncolytic herpes simplex virus to improve systemic delivery

doi: 10.1016/j.omton.2026.201132

Figure Lengend Snippet: Comparison of systemic delivery of FusOn-CD47-luc and FusOn-luc in immune-competent mice pre-immunized with HSV-2 (A) Schematic illustration of the FusOn-series viruses used in this study. The parental FusOn-H2 was generated by replacing the N-terminal domain of the ICP10 gene, which encodes the large subunit of ribonucleotide reductase (RR), with GFP. The locations of glycoprotein C (gC), the terminal repeat long (TR L ) and short (TR S ) regions, and the internal repeats (IR) are indicated. FusOn-luc was constructed by inserting a luciferase gene cassette ( luc ) upstream of the gC locus, whereas FusOn-CD47-luc was generated by fusing the extracellular domain (ECD) of CD47 to gC and inserting the luc cassette at the same position. (B) IVIS imaging of virus distribution following systemic delivery. Balb/c mice were first immunized twice with a gH-deleted infectious single-cycle HSV-2 (DISC-HSV2) before implantation with CT26 tumor cells in the right flank. Once tumors reached approximately 8 mm in diameter, mice received one of the three viruses via tail vein injection at a dose of 2 × 10 6 PFU. Bioluminescence imaging was performed using an IVIS imager on the indicated days post-injection. The locations of the liver and tumor are indicated by red arrows. Representative images from one of five mice in each treatment group are shown.

Article Snippet: African green monkey kidney (Vero) cells, CT26 murine colon cancer cells, LL/2 murine lung cancer cells, and HCT116 human colorectal cancer cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Comparison, Generated, Construct, Luciferase, Imaging, Virus, Injection

Journal: Molecular Therapy Oncology

Article Title: Strategically engineering an oncolytic herpes simplex virus to improve systemic delivery

doi: 10.1016/j.omton.2026.201132

Figure Lengend Snippet: Tumor delivery efficiency of FusOn-SD following systemic delivery in vivo (A) Sequential images of one representative mouse from each group (five mice per group) at the indicated time points after systemic administration of FusOn-SD in immune-competent, CT26-tumor-bearing Balb/c mice pre-immunized with HSV-2. The experimental procedure was identical to that in B. (B) Effect of adoptively transferred human anti-HSV-2 sera on the systemic delivery of FusOn-SD to xenografted human tumors. Mpanc-96 human pancreatic cancer cells were implanted in the right flank of immunodeficient mice. Once tumors reached an approximate size of 8 mm in diameter, mice received an adoptive transfer of 100 μL of either a mixture of eight human anti-HSV-2 sera or non-immune sera as a control, followed by tail vein injection of 2 × 10 6 PFU FusOn-SD. Shown are IVIS images taken 48 h after virus administration, with the tumor sites and corresponding bioluminescent signals highlighted by red circles.

Article Snippet: African green monkey kidney (Vero) cells, CT26 murine colon cancer cells, LL/2 murine lung cancer cells, and HCT116 human colorectal cancer cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: In Vivo, Adoptive Transfer Assay, Control, Injection, Virus

Journal: Molecular Therapy Oncology

Article Title: Strategically engineering an oncolytic herpes simplex virus to improve systemic delivery

doi: 10.1016/j.omton.2026.201132

Figure Lengend Snippet: In vivo evaluation of the antitumor effect of FusOn-SD in immune syngeneic tumor models in immune-competent animals (A) Evaluation of FusOn-SD in the murine CT26 colon cancer model. Immune-competent Balb/c mice were immunized with HSV-2 before CT26 cells were implanted subcutaneously. Oncolytic viruses were given intratumorally at a dose of 2 × 10 6 PFU. Tumor size was measured at the indicated time points and plotted. ★ p < 0.05 compared with other oncolytic viruses and PBS; p < 0.05 compared with PBS. (B) Evaluation of FusOn-SD in the murine LL/2 lung cancer model. Immune-competent C57BL6 mice were immunized with HSV-2 before LL/2 cells were implanted subcutaneously. When tumor became palpable, 2 × 10 6 PFU of the indicated oncolytic viruses were given via the tail vein, either alone or in combination with CP and/or PD1 mAb (detailed treatment schemes are provided in the section). Tumor size was measured at the indicated time points and plotted. Due to the rapid growth of these two tumor models in the control group, the experiments were terminated early to address ethical concerns for animal welfare. ★ p < 0.05 compared with other treatment groups and PBS; p < 0.05 compared with PBS.

Article Snippet: African green monkey kidney (Vero) cells, CT26 murine colon cancer cells, LL/2 murine lung cancer cells, and HCT116 human colorectal cancer cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: In Vivo, Control

Journal: Aging Cell

Article Title: Decreased Glucose Metabolism and Declined Chaperones Are Unique Features Required for the Survival of Senescent Fibroblasts and Pyruvate Dehydrogenase Is a Potent Senolytic Target

doi: 10.1111/acel.70434

Figure Lengend Snippet: Inhibiting PDH, GLS1 and Hsp90 by the combination of CPI‐613+BPTES+17‐AAG gave rise to enhanced senolysis on senescent fibroblasts as well as the therapy‐induced senescent tumor cells. (A, B) The effects of CPI‐613+BPTES+17‐AAG combination treatment on proliferating (A) and senescent (B) BJ cells. For the dose of each compound in use, see the results 2.7 section for more details. ** p < 0.01 by Student's t ‐test. (C, D) The effects of CPI‐613+BPTES+17‐AAG combination treatment on proliferating and Dox‐induced senescent lung adenocarcinoma A549 cells. *** p < 0.001 by Student's t ‐test. (E, F) The effects of CPI‐613+BPTES+17‐AAG combination treatment on proliferating and Dox‐induced senescent cervical carcinoma HeLa cells. *** p < 0.001 by Student's t ‐test. (G) The morphological changes of senescent BJ induced by IR, senescent A549 and HeLa cells induced by Dox at the indicated time of CPI‐613+BPTES+17‐AAG treatment under the light microscopy. The senescent cells without treatment were stained with SA‐β‐gal. Cells were imaged at magnification 200×. (H) The schematic summarization of our findings. The activities of TCA cycle and chaperones are reduced in DNA damage induced senescent cells. Co‐inhibiting Hsp90 and TCA cycle with 17‐AAG+CPI‐613+BPTES combination leads to enhanced selective elimination of senescent cells, hinting TCA cycle and glutaminolysis are novel and potent targets for senolysis.

Article Snippet: Human fibroblasts BJ (RRID: CVCL_3653; ATCC Cat#CRL‐2522), IMR‐90 (RRID: CVCL_0347; ATCC Cat#CCL‐186), WI‐38 (RRID: CVCL_0579; ATCC Cat#CCL‐75) and human cancer cell lines HeLa (RRID: CVCL_0030; ATCC Cat#CRM‐CCL‐2), A549 (RRID: CVCL_0023; ATCC Cat#CRM‐CCL‐185), and U2OS (RRID: CVCL_0042; ATCC Cat#HTB‐96) were purchased from American Type Culture Collection (ATCC) and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 1% penicillin/streptomycin at 37°C under 5% CO 2 .

Techniques: Light Microscopy, Staining

Journal: Aging Cell

Article Title: Decreased Glucose Metabolism and Declined Chaperones Are Unique Features Required for the Survival of Senescent Fibroblasts and Pyruvate Dehydrogenase Is a Potent Senolytic Target

doi: 10.1111/acel.70434

Figure Lengend Snippet: Distinct proteomic and transcriptional signatures of metabolic enzymes and chaperones between senescent fibroblasts and the therapy‐induced senescent tumor cells. (A) The abundance of glycolysis‐related enzymes PFKP, ALDOA, PKM2, TCA cycle‐related PDHA, and glutaminolysis‐related GLS1 was decreased significantly in senescent BJ and IMR‐90 cells compared to their proliferating counterparts. (B) The protein levels of chaperones TCP1, Hsp70, and Hsp90 were decreased significantly in senescent BJ and IMR‐90 cells. (C) The abundance of those glycolysis‐related enzymes remained unchanged or even elevated in Dox‐induced senescent A549, HeLa, and U2OS tumor cells compared to their proliferating counterparts. (D) The abundance of these chaperone proteins in Dox‐induced senescent A549, HeLa, and U2OS tumor cells remained nearly unchanged. The relative abundance of each protein was quantified by signal density scanning on Western blots and normalized to the signal of β‐Actin or β‐tubulin. * p < 0.05, ** p < 0.01 tested by Student's t ‐test.

Article Snippet: Human fibroblasts BJ (RRID: CVCL_3653; ATCC Cat#CRL‐2522), IMR‐90 (RRID: CVCL_0347; ATCC Cat#CCL‐186), WI‐38 (RRID: CVCL_0579; ATCC Cat#CCL‐75) and human cancer cell lines HeLa (RRID: CVCL_0030; ATCC Cat#CRM‐CCL‐2), A549 (RRID: CVCL_0023; ATCC Cat#CRM‐CCL‐185), and U2OS (RRID: CVCL_0042; ATCC Cat#HTB‐96) were purchased from American Type Culture Collection (ATCC) and cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 1% penicillin/streptomycin at 37°C under 5% CO 2 .

Techniques: Western Blot

Journal: Molecular Therapy Oncology

Article Title: Recombinant CALR polarizes and activates macrophages in tumors

doi: 10.1016/j.omton.2025.201121

Figure Lengend Snippet: Recombinant CALR reduces tumor growth and increases M1 macrophages (A) To measure the effect on tumor growth and immune cell activation, bacterial lysate was injected into tumor-bearing mice. Tumors were formed by subcutaneously injecting CT26 murine colon carcinoma cells into the flank of BALB/c mice. After 2 weeks, the mice were injected intratumorally with saline, bacterial control lysate, or lysate from CALR-expressing bacteria. One set of mice received injections at days 0, 3, and 6; were monitored for tumor growth; and their tumors were harvested at day 9 for analysis of immune cells. A second set of mice received only one injection at day 0, and tumors were harvested at day 3 for analysis. (B) Intratumoral injection of CALR lysate decreased tumor growth compared to saline controls ( p = 0.0089). Bacterial control lysate also reduced tumor growth compared to controls ( p = 0.0204). Volumes are reported relative to those on day 0. (C–F) On day 3, recombinant CALR did not affect (C) the number of leukocytes, (D) the number of M1 macrophages (per 10,000 cells analyzed), or the number of M1 macrophages expressing either (E) CD80 or (F) CD86 (per 10,000 cells analyzed) in tumors. (G) On day 9, injection with CALR lysate significantly increased the number of leukocytes in tumors compared to saline controls ( p = 0.0042). (H) On day 9, CALR lysate also significantly increased the number of M1 macrophages in tumors compared to bacterial controls ( p = 0.0480) and saline ( p = 0.0061). (I) CALR lysate significantly increased the number of M1 macrophages expressing CD80 (per 10,000 cells analyzed) compared to saline controls ( p = 0.0063). (J) CALR lysate also increased the number of M1 macrophages expressing CD86 (per 10,000 cells analyzed) compared to bacterial controls ( p = 0.0445) and saline ( p = 0.0077). Data are represented as mean ± SEM. The statistical comparisons in (B) are two-way ANOVA followed by Tukey’s method. The statistical comparisons in (C–J) are ANOVA followed by Tukey’s method. Asterisks indicate significance: ∗ p < 0.05; ∗∗ p < 0.01.

Article Snippet: JAWSII murine dendritic cells and CT26 murine colon carcinoma cells were obtained from ATCC, confirmed by STR profiling by Charles River Research Animal Diagnostic Services, and passaged for fewer than 6 months.

Techniques: Recombinant, Activation Assay, Injection, Saline, Control, Expressing, Bacteria

Journal: Molecular Therapy Oncology

Article Title: Recombinant CALR polarizes and activates macrophages in tumors

doi: 10.1016/j.omton.2025.201121

Figure Lengend Snippet: Recombinant CALR increases helper T cell activity in tumors (A) The extent of T cell infiltration was determined in mice (see ) with CT26 tumors that were intratumorally injected (on days 0, 3, and 6) with saline (PBS), bacterial control lysate (BC), or lysate from CALR-expressing bacteria (CALR). On day 9, injection of CALR lysate significantly increased the number of T cells in tumors (per 10,000 cells analyzed) compared to saline controls ( p = 0.0495). (B) The percentage of helper T cells per 10,000 cells analyzed. (C) On day 9, injection with CALR lysate significantly increased the number of activated helper T cells (per 10,000 cells analyzed) in tumors compared to saline controls ( p = 0.0067). Data are represented as mean ± SEM. The statistical comparisons in (A–C) are ANOVA followed by Dunnett’s test. Asterisks indicate significance: ∗ p < 0.05; ∗∗ p < 0.01.

Article Snippet: JAWSII murine dendritic cells and CT26 murine colon carcinoma cells were obtained from ATCC, confirmed by STR profiling by Charles River Research Animal Diagnostic Services, and passaged for fewer than 6 months.

Techniques: Recombinant, Activity Assay, Injection, Saline, Control, Expressing, Bacteria