Irina Larionova

Background. The treatment efficacy of stage I-II cervical cancer (CC) remains suboptimal due to the risk of recurrence and metastasis, which complicates the selection of optimal treatment strategies and necessitates the search for prognostic markers. This study aimed to identify molecular characteristics and markers associated with progressive stage I-II CC. Material and Methods. Clinical and morphological analyses and whole-transcriptome sequencing were performed, and publicly available data from the Cancer Genome Atlas (TCGA) database were used to validate the sequencing results. Results. Patients with progressive disease demonstrated activation of oncogenic (TNF-α via NF-κB, PI3K/AKT/mTOR, MTORC1, and KRAS), metabolic (oxidative phosphorylation, glycolysis, and hypoxia), immune (TGF-β, responses to IFNγ and IFNα, and IL2/STAT5), and G2/M cell cycle (through regulation of the E2F and MYC transcription factor targets) signaling pathways. the ITGA6 gene expression level was higher in tumors from patients with cervical cancer progression than in patients without cancer progression. Conclusion. The progression of stage I-II cervical cancer is driven by a complex interplay of proliferative, metabolic, and immunosuppressive factors, with ITGA6 identified as a potential marker.  

Tumor is complex tissue where cancer cell fate strongly depends of multicellular component of the tumor microenvironment (TME). The principal control of cancer cell proliferation, oxygen and nutrient supply, and metastatic potential is provided by intratumor immune components. Cancer cells develop multiple strategies to program intratumor immunity, and to escape the host defense. Single-cell RNA-sequencing (scRNA-seq) methods enable identification of singe cell transcriptome of individual cells in the tumor tissue. However, scRNA-seq analyses single cells in suspension where tumor architecture is damaged. In order to identify the essential and potentially targetable molecular interaction of cancer cells and immune cells in the TME, analysis of the single cell transcriptome in the spatial context is critical. Spatial transcriptomics (ST) methods proved the necessary precision for the understanding tumor tissue architecture and heterogeneity, and allow to examine intercellular molecular communications in the TME. In our review, we have summarized the results of the studies which applied the most popular and commercially available ST technologies in cancer research (10x Genomics Visium, 10x Genomics Xenium, Nanostring GeoMx-DSP, Nanostring CosMx, STOmics, and Vizgen MERSCOPE). We elucidated how these studies were able to decipher the new molecular mechanisms of cancer progression controlled by the immune components of the TME. We focused on the new venues for anti-cancer therapy based on immune cell transcriptomics. We discussed the benefits and limitations of the currently used ST approaches. We also summarized how ST technologies can contribute valuable findings for the tissue and cell atlases of cancer.

Immunotherapy is a promising treatment strategy for treating colorectal cancer (CRC). Despite significant advances in this field, resistance and low efficacy of immunotherapy remain a principal problem. One of the most important factors affecting the response to immunotherapy is the tumor microenvironment (TME). Among the components of the TME, tumor-associated macrophages (TAMs) are key immune cells involved in cancer progression by stimulating tumor cell proliferation, angiogenesis, epithelial-mesenchymal transition, metastasis, and tumor immune evasion. This review presents currently investigated combination therapy based on the immune checkpoint inhibitors and inhibitors of diverse components of the TME, including TAMs, that can potentially increase the effectiveness of CRC treatment. Therapeutic efficacy, together with the functional activity of TAMs, is estimated in multiple preclinical data obtained with diverse in vitro and in vivo models. Ongoing clinical trials demonstrated the association of treatment effectiveness with TAM phenotypes and functions.

Colorectal cancer is the third most common cancer worldwide, making lymph node recovery critical for treatment decisions and prognosis. Within the colorectal tumor microenvironment, the metabolic programming of tumor-associated macrophages (TAMs) can drive both pro- and anti-tumor responses, yet the specific glycolytic pathways governing their pro-metastatic conversion present promising therapeutic targets. This study investigated the role of glycolysis activating enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) in mediating TAMs metabolic polarization, and its potential as a therapeutic target. PFKFB3 expression was found to be predominant in TAMs in CRC tumor samples. Lipidomic analysis performed by HPLC-MS/MS revealed that PFKFB3 inhibition altered glycerophospholipid metabolism (p = 6.13 × 10⁻¹⁰), and shifted TAMs toward sphingolipid-mediated immunosuppressive metabolism. PFKFB3 activity was associated with a specific reduction in asparagine availability, potentially pointing to a targeted reprogramming of amino acid metabolism supporting distinct TAM functions under conditions of intra-tumoral metabolic stress. These findings highlight PFKFB3 as an essential regulator of TAMs pro-tumoral metabolism in CRC, particularly in colon cancer.

BACKGROUND: The spatial organization of immune cells within tumors is an important area of investigation in studies of interactions between tumor cells and the tumor microenvironment. This is particularly relevant because some immune cells function through direct contact with their target cells, whereas others communicate over a distance via paracrine signaling involving cytokines. Thus, the topography of tumor cells and microenvironmental cells may determine the possibility and nature of intercellular interactions and thereby influence the functional effects of immune cells. AIM: This study aimed to compare the spatial transcriptomic profiles of tumor and stromal regions enriched in macrophages in triple-negative breast cancer. METHODS: Eight patients with triple-negative breast cancer were included. Spatial transcriptomic analysis was performed on formalin-fixed, paraffin-embedded tissue sections using high-throughput RNA sequencing with the 10X Visium platform. The annotated spots enriched in intraepithelial and stromal macrophages were used for downstream bioinformatic analysis. RESULTS: A total of 437 differentially expressed genes were identified between the two groups of spots containing macrophages with distinct spatial localization. Spots with intraepithelial macrophages were characterized by activation of processes related to cytokine and chemokine signaling, regulation of regulatory T-cell differentiation, organization of cell–cell contacts, wound healing, and inhibition of viral activity. Spots enriched in stromal macrophages demonstrated activation of biological processes associated with the regulation of angiogenesis, cell migration and recruitment, cell adhesion, and stromal remodeling. CONCLUSION: Macrophage topography within primary tumors of triple-negative breast cancer is associated with their functional characteristics. These fundamental findings may be useful for developing prognostic criteria and therapeutic approaches aimed at modulating the tumor microenvironment to improve long-term outcomes in patients with triple-negative breast cancer.

Colorectal cancer is the third most common cancer worldwide, making lymph node re-covery critical for treatment decisions and prognosis. Within the colorectal tumor mi-croenvironment, the metabolic programming of tumor-associated macrophages (TAMs) can drive both pro- and anti-tumor responses, yet the specific glycolytic pathways gov-erning their pro-metastatic conversion present promising therapeutic targets. This study investigated the role of glycolysis activating enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) in mediating TAM metabolic polarization, and its potential as a therapeutic target. PFKFB3 expression was found to be predominant in tumor-associated macrophages in CRC human tumor sam-ples. Lipidomic analysis performed by HPLC-MS/MS revealed that PFKFB3 inhibition altered glycerophospholipid metabolism (p=6.13E-10), and shifted TAMs toward sphingolipid-mediated immunosuppressive metabolism. PFKFB3 activity was associated with a specific reduction in asparagine availability, potentially pointing to a targeted reprogramming of amino acid metabolism supporting distinct macrophage functions under conditions of intra-tumoral metabolic stress. These findings highlight PFKFB3 as an essential metabolic regulator of TAMs pro-tumoral activity in CRC, particularly in colon cancer.

The purpose of the study was to analyze the 2-year survival rates in colorectal cancer (CRC) patients with synchronous liver metastases after combined modality treatment including perioperative chemotherapy.Material and Methods. A pilot prospective study included 30 colorectal cancer patients with initially resectable liver metastases, who received combined modality treatment including 3 courses of neoadjuvant chemotherapy with FOLFOXIRI in combination with cetuximab or bevacizumab (depending on the mutational status of the RAS family genes), radical surgery and 3 courses of adjuvant chemotherapy with FOLFOXIRI. The 2-year survival rates in colorectal cancer with synchronous liver metastases were analyzed on the basis of the main clinical and morphological characteristics of the tumor.Results. Within 2-years of follow-up, disease progression occurred in 7 (23.3 %) patients, including liver metastases in 4 (13.3 %) and multiple lung metastases in 3 (10 %) patients. There were no relapses in the area of intestinal and liver resection. One patient (3.3 %) died from disease progression. The 2-year recurrence-free and overall survival rates were 76.7 % and 96.7 %, respectively. The survival rate was significantly lower in patients with урТ4 primary tumor than in patients with урT3 tumor (44.4 % and 88.2 %, respectively, p=0.015). In patients without lymph node metastases ypN0, the survival rate was 100 %, and in patients with ypN2, the survival rate decreased to 50 % (p=0.016). The survival rate reached 94.1 % in patients with 1–3 liver metastases, and it was 50 % in patients with 7–11 metastases (p=0.02).Conclusion. Perioperative chemotherapy in CRC patients with initially resectable synchronous liver metastases provides encouraging 2-year survival rates, which are directly dependent on the extent of the tumor.

Background/Objectives: In gastric cancer, only a subset of patients benefit clinically from neoadjuvant chemoimmunotherapy, underscoring the need for robust biomarkers that can predict treatment responses and guide personalized immunotherapy. This study aimed to characterize the immune microenvironment of gastric tumors and identify predictive markers associated with therapeutic efficacy. Methods: We prospectively enrolled 16 patients with histologically confirmed, PD-L1–positive (CPS ≥ 1) gastric adenocarcinoma (T_2–4N_0–1M₀). All patients received eight cycles of FLOT chemotherapy combined with pembrolizumab. Treatment response was assessed by Mandard tumor regression grading. Spatial transcriptomic profiling (10x Genomics Visium) and multiplex immunofluorescence were used to evaluate tumor-infiltrating immune cell subsets and PD-1 expression at baseline and after treatment. Results: Transcriptomic analysis differentiated the immune landscapes of responders from non-responders. Responders exhibited elevated expression of IL1B, CXCL5, HMGB1, and IFNGR2, indicative of an inflamed tumor microenvironment and type I/II interferon signaling. In contrast, non-responders demonstrated upregulation of immunosuppressive genes such as LGALS3, IDO1, and CD55, along with enrichment in oxidative phosphorylation and antigen presentation pathways. Multiplex immunofluorescence confirmed a higher density of FoxP3⁺ regulatory T cells in non-responders (median 5.36% vs. 2.41%; p = 0.0032). Notably, PD-1⁺ CD8⁺ T cell and PD-1⁺ FoxP3⁺ Treg frequencies were significantly elevated in non-responders, suggesting that PD-1 expression within cytotoxic and regulatory compartments may contribute to immune evasion. No substantial differences were observed in PD-L1 CPS or PD-1⁺ B cells and PD-1⁺ macrophages. Conclusions: Our findings identify PD-1⁺ CD8⁺ T cells and PD-1⁺ FoxP3⁺ Tregs as potential biomarkers of resistance to neoadjuvant chemoimmunotherapy in gastric cancer. Transcriptional programs centered on IL1B/CXCL5 and LGALS3/IDO1 define distinct immune phenotypes that may guide future combination strategies targeting both effector and suppressive arms of the tumor immune response.

AIM: to assess early efficacy and tolerability of preoperative chemotherapy combined with targeted therapy in patients with rectal cancer (RC).PATIENTS AND METHODS: a pilot prospective study including 22 RC patients with KRAS wild-type gene is ongoing from 2021. There are 13 (59.1%) females and 9 (40.9%) males. Stage II RC was diagnosed in 2 (9.1%) patients and stage III RC in 20 (90.9%) patients. All patients received 6 cycles of mFOLFOX 6 chemotherapy combined with cetuximab targeted therapy followed by surgery. After completion of surgical treatment, patients with T4 and/or N + received adjuvant chemotherapy for 6 months (taking into account the time of preoperative treatment).RESULTS: the completion rate of preoperative treatment was 90.9%, and a 15% reduction in drug dosages was required in 9.1% of patients. Adverse events were observed in 45.4% of patients. Grade III toxicity was noted in 13.6% of patients (neutropenia in 9.1% and skin rash in 4.5%). The clinical and radiological response to preoperative chemo-targeted therapy was 77.3%, including complete (9.1%) and partial (68.2%) tumor regression. All patients underwent radical surgery. The rate of postoperative complications was 13.6%. Grade I and grade IIIa complications (according to Сlavien-Dindo classification) were observed in 9.1% and 4.5% of cases. Pathological complete response (pCR, according to Mandard TRG 1) was achieved in 13.6% of patients, and good pathological response (TRG 1-2) was observed in 31.8% of patients.CONCLUSION: preoperative chemotherapy combined with targeted therapy in RC patients with the wild type of the KRAS gene has a pronounced damaging effect on the tumor, and an acceptable toxicity profile does not affect intraand postoperative period. The short-term outcomes have been found to be encouraging. The study is ongoing to analyze the survival of patients.

Colorectal cancer (CRC) is a prevalent malignancy with high incidence and mortality rates worldwide. Metabolic disorders, such as obesity, dyslipidemia, hyperglycemia, and hypertension, have been identified as significant risk factors contributing to the development and progression of CRC. These metabolic conditions often result in low-grade inflammation, which plays a crucial role in tumor initiation and progression. Monocytes, which differentiate into tumor-associated macrophages (TAMs) within the tumor microenvironment (TME), are pivotal in mediating these inflammatory responses. Monocytes in the individuals with metabolic disorders exhibit altered phenotypes and functions, enhancing their pro-inflammatory and tumor-promoting activities. Our review describes the intricate links between metabolic disorders and CRC on the level of circulating monocytes, highlighting how metabolic conditions can drive CRC initiation and aggravate tumor progression via programming of immunity. Various monocyte subsets, their phenotypic changes, and their impact on CRC are elucidated, providing insights into potential therapeutic targets for CRC prevention and treatment. Understanding the interplay between metabolic alterations and programming of monocytes-derived TAMs in the TME is essential for developing effective strategies against CRC.