Characterization of Glycolysis-Associated Molecules in the Tumor Microenvironment Revealed by Pan-Cancer Tissues and Lung Cancer Single Cell Data

Altered metabolism is a hallmark of cancer and glycolysis is one of the important factors promoting tumor development. There is however still a lack of molecular characterization glycolysis and comprehensive studies related to tumor glycolysis in the pan-cancer landscape. Here, we applied a gene expression signature to quantify glycolysis in 9229 tumors across 25 cancer types and 7875 human lung cancer single cells and verified the robustness of signature using defined glycolysis samples from previous studies. We classified tumors and cells into glycolysis score-high and -low groups, demonstrated their prognostic associations, and identified genome and transcriptome molecular features associated with glycolysis activity. We observed that glycolysis score-high tumors were associated with worse prognosis across cancer types. High glycolysis tumors exhibited specific driver genes altered by copy number aberrations (CNAs) in most cancer types. Tricarboxylic acid (TCA) cycle, DNA replication, tumor proliferation and other cancer hallmarks were more active in glycolysis-high tumors. Glycolysis signature was strongly correlated with hypoxia signature in all 25 cancer tissues (r > 0.7) and cancer single cells (r > 0.8). In addition, HSPA8 and P4HA1 were screened out as the potential modulating factors to glycolysis as their expression were highly correlated with glycolysis score and glycolysis genes, which enables future efforts for therapeutic options to block the glycolysis and control tumor progression. Our study provides a comprehensive molecular-level understanding of glycolysis with a large sample data and demonstrates the hypoxia pressure, growth signals, oncogene mutation and other potential signals could activate glycolysis, thereby to regulate cell cycle, energy material synthesis, cell proliferation and cancer progression

Single-cell transcriptome analysis reveals the regulatory effects of artesunate on splenic immune cells in polymicrobial sepsis

Sepsis is characterized by a severe and life-threatening host immune response to polymicrobial infection accompanied by organ dysfunction. Studies on the therapeutic effect and mechanism of immunomodulatory drugs on the sepsis-induced hyperinflammatory or immunosuppression states of various immune cells remain limited. This study aimed to investigate the protective effects and underlying mechanism of artesunate (ART) on the splenic microenvironment of cecal ligation and puncture-induced sepsis model mice using single-cell RNA sequencing (scRNA-seq) and experimental validations. The scRNA-seq analysis revealed that ART inhibited the activation of pro-inflammatory macrophages recruited during sepsis. ART could restore neutrophils’ chemotaxis and immune function in the septic spleen. It inhibited the activation of T regulatory cells but promoted the cytotoxic function of natural killer cells during sepsis. ART also promoted the differentiation and activity of splenic B cells in mice with sepsis. These results indicated that ART could alleviate the inflammatory and/or immunosuppressive states of various immune cells involved in sepsis to balance the immune homeostasis within the host. Overall, this study provided a comprehensive investigation of the regulatory effect of ART on the splenic microenvironment in sepsis, thus contributing to the application of ART as adjunctive therapy for the clinical treatment of sepsis

Effects of ovariectomy, estrogen and iCR therapy on body weight, lacrimal gland and submandibular gland weight.

<p>BW, body weight; WGR, weight gain rate; UW, uterus weight; LGW, lacrimal gland weight; LGW/BW final, lacrimal gland and final body weight ratio; SW, submandibular gland weight; SW/BW final, submandibular gland and final body weight ratio. Data are expressed as mean± SEM.</p><p>a means p<0.05 vs. SHAM;</p><p>b means p<0.01 vs. SHAM;</p><p>c means p<0.05 vs. OVX;</p><p>d means p<0.01 vs. OVX;</p><p>e means p<0.01 vs. OVX+E;</p><p>f means p<0.05 vs. OVX+ iCR;</p><p>g means p<0.01 vs. OVX+ iCR.</p><p>Effects of ovariectomy, estrogen and iCR therapy on body weight, lacrimal gland and submandibular gland weight.</p

The percentage of acinar and duct cells area in the lobule of submandibular gland.

<p>A, The percentage of intercellular space area in the lobule of submandibular gland. B, The percentage of acinar cells area in the lobule of submandibular gland. C, The percentage of GCT area in the lobule of submandibular gland. D, The percentage of striated ducts area in the lobule of submandibular gland. a means p<0.01 vs. SHAM; b means p<0.01 vs. OVX; c means p<0.05 vs. OVX; d means p<0.05 vs. OVX+E; e means p<0.05 vs. OVX+ iCR.</p

Cu-Zn SOD expression.

<p>a-e, lacrimal gland; f-j, submandibular gland. Scale bars represent 100 μm (magnification ×400). In the negative control, no brown precipitation was observed, representing no positive response. In the lacrimal gland, positive staining of Cu-Zn SOD mainly occurred on the basolateral side of the acinar cells, while in the submandibular gland, strong staining was found in the GCT and striated ducts.</p

The GCT in the submandibular gland.

<p>A, SHAM; B, OVX; C, OVX + E; D, OVX + iCR. Scale bars represent 1 μm. In the SHAM group, the cell plasma was filled with a high density of secretory granules, and the nucleus was located on the basolateral side of the cell with normal chromatin morphology, and mitochondria were intact. In the OVX group, many of the mitochondria were swollen, with double membrane structure damage and crest fracture, and “balloon-like” mitochondria were common. The condensed chromatin morphology was inside the cells, probably resulted- from the mutual ingestion of apoptotic cells. The color density of the secretory granules was reduced, with a gray appearance. In the OVX+ E and OVX+ iCR groups, most of the mitochondrial structure was normal, the distribution of chromatin was uniform, and the density of black secretory granules was similar to that of the SHAM group.</p

The apoptosis cell of OVX group in lacrimal gland.

<p>The apoptosis cells were seen in the OVX group, which had an irregular nuclear with chromatin pyknosis and lost nuclear membrane. Besides, the endoplasmic reticulum was cracked. The scale bar represents 1μm.</p

The acinar cells in the lacrimal gland.

<p>A, SHAM; B, OVX; C, OVX + E; D, OVX + iCR. All scale bars represent 2 μm. * stand for the lumen. The white arrows indicate the mitochondria lesion, and the black arrow represents the cell membrane sag. Compared with the SHAM group (Fig. 5, A), the endoplasmic reticulum was diluted and arranged in a disorderly fashion in the OVX groups. Most of the mitochondria appeared to be swollen, with dissolution and crest fracture. Moreover, the diameter of the white secretory granules seemed decreased obviously. The cell shrink, with sag on the membrane and an increased intercellular space, was often seen (Fig. 5, B). Estrogen treatment did not ameliorate the enlargement of endoplasmic reticulum, but the mitochondria seemed normal—ridge fracture was rarely seen. Similar to OVX group, the diameter of the secretory granules decreased also, but the extent of cell shrink was more slightly (Fig. 5, C). Interestingly, the iCR therapy was shown to ameliorate the expansion of endoplasmic reticulum, but not the mitochondria lesions. The diameter of the secretory granules was analogous to SHAM group, and the cell atrophy was rarely observed (Fig. 5, D).</p

The expression intensity of Cu-Zn SOD in lacrimal and submandibular glands.

<p>A: The mean IOD of Cu-Zn SOD staining in lacrimal gland. B: The mean IOD of Cu-Zn SOD staining in the acinar cells of the submandibular gland. C: The mean IOD of Cu-Zn SOD staining in the GCT of the submandibular gland. D: The mean IOD of Cu-Zn SOD staining in the striated ducts of the submandibular gland. a means p<0.05 vs. OVX; b means p<0.05 vs. OVX+ iCR.</p