T11TS Treatment Augments Apoptosis of Glioma Associated Brain Endothelial Cells, Hint Toward Anti-Angiogenic Action in Glioma

Abstract

Malignant glioma continues to be a clinical challenge with an urgent need for developing curative therapeutic intervention. Apoptosis induction in tumor-associated endothelial cells represent a central mechanism that counteracts angiogenesis in glioma and other solid tumors. We previously demonstrated that intraperitoneal administration of sheep erythrocyte membrane glycopeptide T11-target structure (T11TS) in rodent glioma model inhibits PI3K/Akt pathway and Raf/MEK/ERK signaling in glioma-associated brain endothelial cells. In the present study, we investigated whether T11TS treatment influence apoptosis signaling in vivo in glioma-associated brain endothelial cells. Annexin-V/PI staining showed that T11TS treatment in glioma-induced rats increases apoptosis of glioma-associated endothelial cells within glioma milieu compared to brain endothelial cells in glioma induced and control groups. Flowcytometric JC-1 assay revealed that T11TS administration triggers loss of mitochondrial membrane potential in glioma-associated brain endothelial cells. Flowcytometry, immunoblotting, and in situ immunofluoresecnt imaging were employed to investigate the effect of T11TS on apoptotic regulatory proteins in brain endothelial cells. T11TS treatment-upmodulated expression of p53, Bax, Fas, FasL, and FADD in glioma associated endothelial cells and downregulated Bcl-2 protein. T11TS therapy induced cytochrome-c release into cytosol, activated caspase -9, 8, 3, and cleaved Bid in glioma associated brain endothelial cells. The study demonstrates that T11TS induces apoptosis in glioma-associated brain endothelial cells via p53 accumulation and activation of intrinsic as well as Fas-dependent extrinsic pathway. The pro-apoptotic action of T11TS on glioma-associated endothelial cells provides crucial insight into how T11TS exerts its anti-angiogenic function in glioma. J. Cell. Physiol. 232: 526-539, 2017. © 2016 Wiley Periodicals, Inc.

Repurposing synthetic and natural derivatives induces apoptosis in an orthotopic glioma‐induced xenograft model by modulating WNT/β‐catenin signaling

“…Interestingly, our quadruplecombinatorial treatment (TCNP) had significantly downregulated the levels of BCL-2 and upregulated the expression levels of cyt-c and BAD (Figures 2 and 3). Since BCL-2 is downregulated in the mitochondria, this would have resulted in increased mitochondrial permeability and release of cyt-c. We assume that the released cyt-c would have activated the caspasescascade, as the expression levels of caspase-8 and caspase-3 were significantly upregulated in the quadruple-drug treated groups (Figures 2 and 3), similar to the previous reports wherein the release of cyt-c had upregulated the levels of caspase-3 [107,108]. Thus, our molecular studies emphasized that the quadruplecombinatorial treatment (TCNP) had modulated WNT/βcatenin signaling cascade by induction of apoptosis.…”

T11TS repress gliomagenic apoptosis of bone marrow hematopoietic stem cells

“…Further, it was that induction of G1 cell cycle arrest in glioma cells by T11TS is associated with up‐regulation of Cip1/Kip1 and concurrent downregulation of cyclin D (1 and 3) (Acharya et al, ). The role of T11TS on glioma potentiates anti‐angiogenic machinery and their molecular aspects has recently been unearthed in our laboratory (Bhattacharya et al, , ; Bhattacharya, Chaudhuri, Singh, & Chaudhuri, ; Singh et al, , ).…”

“…The novel glycopeptide T11TS acts differentially on different cell types regarding death and survival of the cells in question. Interestingly, T11TS causes apoptosis of glioma cells (Mukherjee, Ghosh, Sarkar, Mazumdar, Banerjee, et al, ; Mukherjee, Ghosh, Sarkar, Mazumdar, Sarkar, et al, ) and glioma associated endothelial cells (Bhattacharya et al, ), but protects the BMHSCs from apoptosis during glioma growth. These actions of T11TS are all conducive to glioma amelioration.…”

“…T11TS was isolated from sheep erythrocyte membrane using DEAE‐cellulose column chromatography as described previously (Bhattacharjee et al, ; Bhattacharya, Singh, Chaudhuri, Datta, & Chaudhuri, ; Sarkar et al, ). The third eluate fraction exclusively contains T11TS and was selected as the fraction of choice for our experiment.…”

The novel-molecule T11TS facilitated arousal of glioma-mediated dormancy of bone-marrow hematopoietic stem-cells

“…The published literature has also conclusively documented the gliomagenic global immune suppression on gliomabearing animals and the beneficial immunomodulatory effect of T11TS immunotherapy [60][61][62][63] . Following T11TS administration, there is profound immune stimulation which in turn results in the proliferation of the immunocytes and rejuvenates immune suppressive state of the malignant glioma [61,62,[64][65][66][67] . Therefore, the novelty of our work is that this is the first reporting of the status of HSCs during glioma occurrence and thereafter following immunotherapy, which has not been explored so far.…”