Tumor Cell Growth Research Articles

PAMAM dendrimers based co-delivery of methotrexate and berberine for targeting of Hela cancer cells

Polyamidoamine dendrimer (PAMAM) is a class of synthetic macromolecular polymers for targeted drug delivery. PAMAM dendrimers are characterized by a pure defined structure, adjustable nanoscale dimensions, mono-dispersity, and versatile surface modification. The objective of this study was to study the covalent coupling of methotrexate (MTX) to PAMAM dendrimer, which was loaded with the natural product anticancer agent, berberine (BER) for the targeting of HeLa cells. The successful preparation of MTX-conjugated PAMAM loaded with BER (MTX-PAMAM-BER) was confirmed by Fourier transform infrared spectroscopy and particle size was evaluated using dynamic light scattering. The biological assays, MTT, flow cytometry, ROS levels evaluation and DAPI staining were used to assess the cytotoxicity effect of the prepared nanosystem. The findings indicated that MTX-PAMAM-BER exhibited greater suppression of tumor cell growth in comparison to BER, MTX, PAMAM-BER, and MTX-PAMAM. Besides, the noteworthy ROS level has been seen in the treated cells with MTX-PAMAM-BER. Finally, it should be stated that the fabricated MTX-PAMAM-BER co-delivery nanosystem could be a promising agent for cancer therapy and targeting.

MiR-4448/Girdin/Akt/AMPK axis inhibits EZH2-mediated EMT and tumorigenesis in small-cell lung cancer.

Small-cell lung cancer (SCLC) shows high enhancer of zeste homolog 2 (EZH2) expressions. EZH2-mediated epigenetics promote epithelial-mesenchymal transition (EMT), enhancing invasive and metastatic potential in malignancies. MicroRNAs (miRNAs), small noncoding RNAs, modulate EMT, determining tumor phenotypes. However, the association between miRNAs and EZH2 in SCLC remains to be clarified-we aimed to identify a novel tumorigenic mechanism through miRNAs, EZH2, and EMT in SCLC, leading to future therapeutic applications. We analyzed EZH2 and E-cadherin expressions in lung cancer cell lines and tumor tissues from 34 SCLC patients and confirmed EZH2 siRNA-mediated EMT inhibition. miRNA expression profiles were compared between EZH2 knockdown SCLC cells and negative control SCLC cells using miRNA array. We identified a target miRNA of EZH2 showing expressional differences in EZH2-knockdown cells and analyzed the impact of the miRNA on EZH2-mediated EMT and tumorigenesis. All SCLC cells showed increased EZH2 and decreased E-cadherin expressions. SCLC tissues had higher EZH2 and lower E-cadherin expressions than other lung cancer tissues. miRNA array revealed that miR-4448 expression increased in EZH2-knockdown SCLC cells. miR-4448 overexpression reduced tumor cell growth and prevented EMT. miR-4448 bound to the 3'UTR of the girdin gene and suppressed its expression, thereby decreasing Akt phosphorylation at Ser473. Attenuated Akt phosphorylation resulted in AMP-activated protein kinase (AMPK) phosphorylation at Thr172 and 183, enhancing EZH2 phosphorylation at Thr311. SCLC characterized high EZH2 expression and promoted EMT, compared with non-small cell lung cancer. miR-4448 inhibited Girdin expression, reducing Akt phosphorylation, and enhancing AMPK and EZH2 phosphorylation. Eventually, miR-4448 prevented EZH2-mediated EMT and tumorigenesis by modulating the Girdin/Akt/AMPK axis in SCLC. miR-4448 might be a potential SCLC inhibitor.

Purines and purinergic receptors in primary tumors of the central nervous system.

Purine nucleotides and nucleosides play critical roles in various pathological conditions, including tumor cell growth. Adenosine triphosphate (ATP) activates pro-tumor receptors, while adenosine (ADO) is a potent immunosuppressant and modulator of cell growth. This study aims to analyze the purinergic actions of ATP and its metabolites, associated enzymes, and P1 or P2 class receptors in primary central nervous system tumors. Additionally, we sought to correlate the levels of nucleosides and the density of P1, P2X, and P2Y receptors in cells with tumor progression. The results indicate that purinergic signaling depends on the receptor concentration and signaling molecules specific to each cell type, tissue, and tumor histology. The purinergic system may function as either a tumor-promoting agent or an antitumor factor, depending on the microenvironmental conditions and the concentrations of receptors and their respective activators. Notably, ATP emerges as the most significant extracellular signal, capable of being converted into other cellular stimulators pertinent to neoplasms, such as adenosine diphosphate, adenosine monophosphate, adenosine, and inosine. Consequently, a cascade of responses to these stimuli promotes tumor development, cell division, and metastasis. Purine nucleotides in central nervous system tumors are pivotal in cellular responses in glioblastoma multiforme, vestibular schwannoma, medulloblastoma, adenomas, gliomas, meningiomas, and pineal tumors. These findings hold the potential for developing novel therapeutic strategies and aiding in therapeutic management.

Gremlin1: a BMP antagonist with therapeutic potential in Oncology.

Gremlins, originating from early 20th-century Western folklore, are mythical creatures known for causing mechanical malfunctions and electronic failures, aptly dubbed "little devils". Analogously, GREM1 acts like a horde of these mischievous entities by antagonizing the bone morphogenetic protein (BMP signaling) pathway or through other non-BMP dependent mechanisms (such as binding to Fibroblast Growth Factor Receptor 1and Epidermal Growth Factor Receptor) contributing to the malignant progression of various cancers. The overexpression of GREM1 promotes tumor cell growth and survival, enhances angiogenesis within the tumor microenvironment, and creates favorable conditions for tumor development and dissemination. Consequently, inhibiting the activity of GREM1 or blocking its interaction with BMP presents a promising strategy for suppressing tumor growth and metastasis. However, the role of GREM1 in cancer remains a subject of debate, with evidence suggesting both oncogenic and tumor-suppressive functions. Currently, several pharmaceutical companies are researching the GREM1 target, with some advancing to Phase I/II clinical trials. This article will provide a detailed overview of the GREM1 target and explore its potential role in cancer therapy.

NIR Enhanced pH-Responsive Microneedles for Synergetic Therapy of Melanoma.

Melanoma has emerged as a significant threat to human life and health. Microneedle (MN)-mediated transdermal drug delivery (TDD) has garnered attention in melanoma treatment for bypassing the first-pass effect. However, the propensity of melanoma to metastasize presents substantial challenges for MN mediated local treatment. Developing systemic therapies, such as immunotherapy in combination with TDD, is crucial for achieving effective melanoma treatment. Herein, a polyvinyl alcohol (PVA) MN-mediated multifunctional TDD system, designated MN@PDA@1-MT/CUR/DOX@HA (MN@PMCDH), was developed for synergetic chemotherapy/photothermal/immunotherapy of melanoma. PMCDH nanomedicines penetrate deep skin layers through MNs, accumulate at tumor sites guided by hyaluronic acid (HA), and selectively release drugs in response to the acidic tumor microenvironment and near-infrared (NIR) stimulation. Released curcumin (CUR) significantly enhances the efficacy of photothermal therapy (PTT) and chemotherapy, as well as improves the induction of immunogenic cell death (ICD) by increasing melanoma sensitivity to polydopamine (PDA)-mediated photothermal effects and doxorubicin (DOX). Moreover, the incorporation of 1-methyltryptophan (1-MT) to reverse the tumor immunosuppressive microenvironment can further enhance the effects of immunotherapy. In vitro studies revealed that the MN@PMCDH system can effectively induce ICD and inhibit tumor cell growth. Additionally, remarkable deep tumor cell inhibition effects are also achieved in 3D tumor models.

Epigenetic reader ZMYND11 noncanonical function restricts HNRNPA1-mediated stress granule formation and oncogenic activity

Epigenetic readers frequently affect gene regulation, correlate with disease prognosis, and hold significant potential as therapeutic targets for cancer. Zinc finger MYND-type containing 11 (ZMYND11) is notably recognized for reading the epigenetic marker H3.3K36me3; however, its broader functions and mechanisms of action in cancer remain underexplored. Here, we report that ZMYND11 downregulation is prevalent across various cancers and profoundly correlates with poorer outcomes in prostate cancer patients. Depletion of ZMYND11 promotes tumor cell growth, migration, and invasion in vitro, as well as tumor formation and metastasis in vivo. Mechanistically, we discover that ZMYND11 exhibits tumor suppressive roles by recognizing arginine-194-methylated HNRNPA1 dependent on its MYND domain, thereby retaining HNRNPA1 in the nucleus and preventing the formation of stress granules in the cytoplasm. Furthermore, ZMYND11 counteracts the HNRNPA1-driven increase in the PKM2/PKM1 ratio, thus mitigating the aggressive tumor phenotype promoted by PKM2. Remarkably, ZMYND11 recognition of HNRNPA1 can be disrupted by pharmaceutical inhibition of the arginine methyltransferase PRMT5. Tumors with low ZMYND11 expression show sensitivity to PRMT5 inhibitors. Taken together, our findings uncover a previously unexplored noncanonical role of ZMYND11 as a nonhistone methylation reader and underscore the critical importance of arginine methylation in the ZMYND11-HNRNPA1 interaction for restraining tumor progression, thereby proposing novel therapeutic targets and potential biomarkers for cancer treatment.

Novel Anti-Trop2 Nanobodies Disrupt Receptor Dimerization and Inhibit Tumor Cell Growth.

Background: Trop2 (trophoblast cell-surface antigen 2) is overexpressed in multiple malignancies and is closely associated with poor prognosis, thus positioning it as a promising target for pan-cancer therapies. Despite the approval of Trop2-targeted antibody-drug conjugates (ADCs), challenges such as side effects, drug resistance, and limited efficacy persist. Recent studies have shown that the dimeric forms of Trop2 are crucial for its oncogenic functions, and the binding epitopes of existing Trop2-targeted drugs lie distant from the dimerization interface, potentially limiting their antitumor efficacy. Method: A well-established synthetic nanobody library was screened against Trop2-ECD. The identified nanobodies were extensively characterized, including their binding specificity and affinity, as well as their bioactivities in antigen-antibody endocytosis, cell proliferation, and the inhibition of Trop2 dimer assembly. Finally, ELISA based epitope analysis and AlphaFold 3 were employed to elucidate the binding modes of the nanobodies. Results: We identified two nanobodies, N14 and N152, which demonstrated high affinity and specificity for Trop2. Cell-based assays confirmed that N14 and N152 can facilitate receptor internalization and inhibit growth in Trop2-positive tumor cells. Epitope analysis uncovered that N14 and N152 are capable of binding with all three subdomains of Trop2-ECD and effectively disrupt Trop2 dimerization. Predictive modeling suggests that N14 and N152 likely target the epitopes at the interface of Trop2 cis-dimerization. The binding modality and mechanism of action demonstrated by N14 and N152 are unique among Trop2-targeted antibodies. Conclusions: we identified two novel nanobodies, N14 and N152, that specifically bind to Trop2. Importantly, these nanobodies exhibit significant anti-tumor efficacy and distinctive binding patterns, underscoring their potential as innovative Trop2-targeted therapeutics.

Prohibitin 2 orchestrates long noncoding RNA and gene transcription to accelerate tumorigenesis

The spatial co-presence of aberrant long non-coding RNAs (lncRNAs) and abnormal coding genes contributes to malignancy development in various tumors. However, precise coordinated mechanisms underlying this phenomenon in tumorigenesis remains incompletely understood. Here, we show that Prohibitin 2 (PHB2) orchestrates the transcription of an oncogenic CASC15-New-Isoform 2 (CANT2) lncRNA and the coding tumor-suppressor gene CCBE1, thereby accelerating melanoma tumorigenesis. In melanoma cells, PHB2 initially accesses the open chromatin sites at the CANT2 promoter, recruiting MLL2 to augment H3K4 trimethylation and activate CANT2 transcription. Intriguingly, PHB2 further binds the activated CANT2 transcript, targeting the promoter of the tumor-suppressor gene CCBE1. This interaction recruits histone deacetylase HDAC1 to decrease H3K27 acetylation at the CCBE1 promoter and inhibit its transcription, significantly promoting tumor cell growth and metastasis both in vitro and in vivo. Our study elucidates a PHB2-mediated mechanism that orchestrates the aberrant transcription of lncRNAs and coding genes, providing an intriguing epigenetic regulatory model in tumorigenesis.

Lymphoma cell-driven IL-16 is expressed in activated B-cell-like diffuse large Bcell lymphomas and regulates the pro-tumor microenvironment.

The activated B-cell-like subtype of diffuse large B-cell lymphoma (ABC-DLBCL) displays a worse outcome than the germinal center B-cell-like subtype (GCB-DLBCL). Currently, targeting tumor microenvironment (TME) is the promising approach to cure DLBCL with profound molecular heterogeneity, however, the factors affecting the tumor-promoting TME of ABCDLBCL are elusive. Here, cytokine interleukin-16 (IL-16) is expressed in tumor cells of ABCDLBCL and secreted by the cleavage of active caspase-3. The serum IL-16 levels are not only a sensitive marker of treatment response but also positively correlated with unfavorable prognosis in DLBCL patients. While IL-16 shows few direct promotional effects on tumor cell growth in vitro, its bioactive form significantly promotes tumor progression in vivo. Mechanically, IL-16 increases the infiltration of macrophages by the chemotaxis of CD4+ monocytes in the TME enhancing angiogenesis, and the expression of cytokine IL-6 and IL-10, as well as decreasing T cell infiltration to accelerate tumor progression. This study demonstrates that IL-16 exerts a novel role in coordinating the bidirectional interactions between tumor progression and the TME. IMM0306, a fusion protein of CD20 mAb with the CD47 binding domain of SIRPα, reverses the tumorpromoting effects of IL-16,which provides new insight into treatment strategy in ABC-DLBCL.

DNp73 enhances tumor progression and immune evasion in multiple myeloma by targeting the MYC and MYCN pathways.

Multiple myeloma (MM) is an incurable hematological malignancy with high chromosome instability and heavy dependence on the immunosuppressive bone marrow microenvironment. P53 mutations are adverse prognostic factors in MM; however, clinically, some patients without P53 mutations also exhibit aggressive disease progression. DNp73, an inhibitor of TP53 tumor suppressor family members, drives drug resistance and cancer progression in several solid malignancies. Nevertheless, the biological functions of DNp73 and the molecular mechanisms in myelomagenesis remain unclear. The effects of DNp73 on proliferation and drug sensitivity were assessed using flow cytometry and xenograft models. To investigate the mechanisms of drug resistance, RNA-seq and ChIP-seq analyses were performed in MM cell lines, with validation by Western blot and RT-qPCR. Immunofluorescence and transwell assays were used to assess DNA damage and cell invasion in MM cells. Additionally, in vitro phagocytosis assays were conducted to confirm the role of DNp73 in immune evasion. Our study found that activation of NF-κB-p65 in multiple myeloma cells with different p53 mutation statuses upregulates DNp73 expression at the transcriptional level. Forced expression of DNp73 promoted aggressive proliferation and multidrug resistance in MM cells. Bulk RNA-seq analysis was conducted to assess the levels of MYCN, MYC, and CDK7. A ChIP-qPCR assay was used to reveal that DNp73 acts as a transcription factor regulating MYCN gene expression. Bulk RNA-seq analysis demonstrated increased levels of MYCN, MYC, and CDK7 with forced DNp73 expression in MM cells. A ChIP-qPCR assay revealed that DNp73 upregulates MYCN gene expression as a transcription factor. Additionally, DNp73 promoted immune evasion of MM cells by upregulating MYC target genes CD47 and PD-L1. Blockade of the CD47/SIRPα and PD-1/PD-L1 signaling pathways by the SIRPα-Fc fusion protein IMM01 and monoclonal antibody atezolizumab significantly restored the anti-MM activity of macrophages and T cells in the microenvironment, respectively. In summary, our study demonstrated for the first time that the p53 family member DNp73 remarkably induces proliferation, drug resistance, and immune escape of myeloma cells by directly targeting MYCN and regulating the MYC pathway. The oncogenic function of DNp73 is independent of p53 status in MM cells. These data contribute to a better understanding of the function of TP53 and its family members in tumorigenesis. Moreover, our study clarified that DNp73 overexpression not only promotes aggressive growth of tumor cells but, more importantly, promotes immune escape of MM cells through upregulation of immune checkpoints. DNp73 could serve as a biomarker for immunotherapy targeting PD-L1 and CD47 blockade in MM patients.

Phytochemical and Pharmacological Profile of Plants from Genus Buxus: A Review

Buxus is an evergreen shrub of the Buxaceae family. A comprehensive survey of the literature has been conducted, revealing that alkaloids are the predominant metabolites characterized by their noteworthy biological activities. Of particular interest are the alkaloids that have exhibited significant cytotoxic activity against various cell lines, including HL-60, SMMC-7721, A549, MCF- 7, SW480, ES2, and A2780 cell lines. CVB-D, buxbodine B, and buxmicrophyllines F can inhibit the growth of tumor cells and induce tumor cell apoptosis. This article reviews the chemical constituents, antitumor effect, and mechanism of Buxus alkaloids in recent years so as to provide a reference for its further development and utilization of medicinal plants of the genus Buxus.

Optimized patient-specific immune checkpoint inhibitor therapies for cancer treatment based on tumor immune microenvironment modeling.

Enhancing patient response to immune checkpoint inhibitors (ICIs) is crucial in cancer immunotherapy. We aim to create a data-driven mathematical model of the tumor immune microenvironment (TIME) and utilize deep reinforcement learning (DRL) to optimize patient-specific ICI therapy combined with chemotherapy (ICC). Using patients' genomic and transcriptomic data, we develop an ordinary differential equations (ODEs)-based TIME dynamic evolutionary model to characterize interactions among chemotherapy, ICIs, immune cells, and tumor cells. A DRL agent is trained to determine the personalized optimal ICC therapy. Numerical experiments with real-world data demonstrate that the proposed TIME model can predict ICI therapy response. The DRL-derived personalized ICC therapy outperforms predefined fixed schedules. For tumors with extremely low CD8 + T cell infiltration ('extremely cold tumors'), the DRL agent recommends high-dosage chemotherapy alone. For tumors with higher CD8 + T cell infiltration ('cold' and 'hot tumors'), an appropriate chemotherapy dosage induces CD8 + T cell proliferation, enhancing ICI therapy outcomes. Specifically, for 'hot tumors', chemotherapy and ICI are administered simultaneously, while for 'cold tumors', a mid-dosage of chemotherapy makes the TIME 'hotter' before ICI administration. However, in several 'cold tumors' with rapid resistant tumor cell growth, ICC eventually fails. This study highlights the potential of utilizing real-world clinical data and DRL algorithm to develop personalized optimal ICC by understanding the complex biological dynamics of a patient's TIME. Our ODE-based TIME dynamic evolutionary model offers a theoretical framework for determining the best use of ICI, and the proposed DRL agent may guide personalized ICC schedules.

Abstract C109: Identifying a novel metabolic reprogramming strategy to inhibit diabetes-associated TNBC in African American

Abstract Cancer cells have an increased glucose metabolism, primarily characterized by augmented glucose uptake and aerobic glycolysis, converting glucose into pyruvate, eventually producing lactate. Breast cancer (BC) is the most commonly diagnosed cancer and the second leading cause of death among women in the United States. The triple-negative breast cancer (TNBC) subgroup is the most aggressive, metastatic, and refractory because of the absence of the three characteristic receptors: Estrogen (ER), progesterone (PR), and human epidermal growth factor (Her2/neu), representing approximately 10-15% of all BC cases. Racial disparities analysis demonstrates that African American (AA) has a higher incidence of TNBC subtype, with an earlier onset, more advanced stage, more aggressive histologic features, and poor survival, compared to other ethnic groups. Hyperglycemia affects the cancer progression through metabolic reprogramming and molecular alterations. AA women with Type 2 diabetes (T2D) are at increased risk of developing ER-negative BC. The reasonable combination therapy of metformin with NF-𝜅Bi and MCT4i, is promising because it can rapidly disable tumor cell growth but with a minor effect on normal cells, holding promise as an anti-BC treatment. Because of the potential side effects of MCT4 inhibitors and the potent inhibitory effect of microRNA on target genes, we consider using microRNA instead of inhibitors to suppress MCT4 expression. We will investigate the anti-diabetes-associated TNBC effect of the MRS, consisting of blocking lactate export via MCT4 degradation by miR-206, in combination with promoting lactate production via forcing glycolysis with metformin and NF-𝜅Bi treatment, in a preclinical study of AA women. Moreover, our previous study and other studies have also shown an elevated expression of Metastasis Associated Lung Adenocarcinoma Transcript 1 (MALAT1) in BC tissues and cells, particularly in metastatic TNBC. We will also explore the molecular mechanisms by which high glucose (HG) leads to lactate accumulation in BC cells. Our overall strategy is that excessive glucose uptake upregulates the MCT4 through a MALAT1/miR-206/MCT4 axis, miR-206-directed MRS shifts glucose metabolism from Oxidative phosphorylation (OXPHOS) to glycolysis, increasing lactate production. Meanwhile, blocking the export of excessive lactate through MCT4 degradation causes high intracellular lactate accumulation and a decrease in intracellular pH (pHi), resulting in metabolic crisis and BC cell death. Due to a complex interplay between hyperglycemia and cancer, antihyperglycemic drugs have often been devised in combination with anticancer drugs for cancer treatment. Our proposed approach will broadly impact the field by providing a novel idea for therapies targeting cancer cell lactate metabolism. In the long term, these studies may reveal an effective therapeutic strategy for diabetes-associated BC, especially in AA women. Citation Format: Qiongyu Hao, Yong Wu, Yanyuan Wu, Jaydutt Vadgama. Identifying a novel metabolic reprogramming strategy to inhibit diabetes-associated TNBC in African American [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C109.

Interleukin-15 and high-intensity exercise: relationship with inflammation, body composition and fitness in cancer survivors.

Pre-clinical murine and in vitro models have demonstrated that exercise suppresses tumour and cancer cell growth. These anti-oncogenic effects of exercise were associated with the exercise-mediated release of myokines such as interleukin (IL)-15. However, no study has quantified the acute IL-15 response in human cancer survivors, and whether physiological adaptations to exercise training (i.e. body composition and cardiorespiratory fitness) influence this response. In the present study breast, prostate and colorectal cancer survivors (n=14) completed a single bout of high-intensity interval exercise (HIIE) [4×4min at 85-95% heart rate (HR) peak, 3min at 50-70% HR peak] before and after 7months of three times weekly high-intensity interval training (HIIT) on a cycle ergometer. At each time point venous blood was sampled before and immediately after HIIE to assess the acute myokine (IL-15, IL-6, IL-10, IL-1ra) responses. Markers of inflammation, cardiorespiratory fitness and measures of body composition were obtained at baseline and 7months. An acute bout of HIIE resulted in a significant increase in IL-15 concentrations (pre-intervention: 113%; P=0.013, post-intervention: 102%; P=0.005). Post-exercise IL-15 concentrations were associated with all other post-exercise myokine concentrations, lean mass (P=0.031), visceral adipose tissue (P=0.039) and absolute peak (P=0.032). There was no significant effect of 7months of HIIT on pre- or post-HIIE IL-15 concentrations (P>0.05). This study demonstrates HIIE is a sufficient stimulus to increase circulating IL-15 and other myokines including IL-6, IL-10 and IL-1ra which may be clinically relevant in the anti-oncogenic effect of exercise and repetitive exposure to these effects may contribute to the positive relationship between exercise and cancer recurrence. KEY POINTS: Exercise has been demonstrated to reduce the risk of cancer recurrence. Pre-clinical murine and in vitro models have demonstrated that exercise suppresses tumour and cancer cell growth, mediated by exercise-induced myokines (IL-6 and IL-15). High-intensity interval exercise significantly increased myokines associated with the anti-oncogenic effect of exercise and the magnitude of response was associated with lean mass, but training did not appear to influence this response. Given IL-15 has been implicated in the anti-oncogenic effect of exercise and is being explored as an immunotherapy agent, high-intensity interval exercise may improve outcomes for people living beyond cancer through IL-15-mediated pathways. Interventions that increase lean mass may also enhance this response.

Senolytic effect of triterpenoid complex from Ganoderma lucidum on adriamycin-induced senescent human hepatocellular carcinoma cells model in vitro and in vivo.

Ganoderma lucidum (G. lucidum) is a famous medicinal mushroom that has been reported to prevent and treat a variety of diseases. Different extractions from G. lucidum have been used to manage age-related diseases, including cancer. Nevertheless, the senolytic activity of G. lucidum against senescent cancer cells has not been investigated. Although cellular senescence causes tumor growth inhibition, senescent cells promote the growth of the neighboring tumor cells through paracrine effects. Therefore, the elimination of senescent cells is a new strategy for cancer treatment. In this study, senescence was triggered in HCC cells by the chemotherapeutic agent Adriamycin (ADR), and subsequently, cells were treated with TC to assess its senolytic activity. We found for the first time that the triterpenoid complex (TC) from G. lucidum had senolytic effect, which could selectively eliminate adriamycin (ADR)-induced senescent cells (SCs) of hepatocellular carcinoma (HCC) cells via caspase-dependent and mitochondrial pathways-mediated apoptosis and reduce the levels of senescence markers, thereby inhibiting the progression of cancers caused by SCs. TC could block autophagy at the late stage in SCs, resulting in a significant activation of TC-induced apoptosis. Furthermore, TC inhibited the senescence-associated secretory phenotype (SASP) in SCs through the inhibition of NF-κB, TFEB, P38, ERK, and mTOR signaling pathways and reducing the number of SCs. Sequential administration of ADR and TC in vivo significantly reduced tumor growth and reversed the toxicity of ADR. A triterpenoid complex isolated from G. lucidum may serve as a novel senolytic agent against SCs, and its combination with chemotherapeutic agents may enhance their antitumor efficacy.

Intravenous leiomyomatosis: A rare case report

Introduction: Intravascular leiomyomatosis (IVL) is a rare histological benign tumor without invading the tissue of the venous vessels themselves. It is characterized by continuous intraluminal growth of benign smooth muscle tumor cells along with the uterine vein to the pelvic and abdominal venous systems, including the iliac vein and inferior vena cava, and even to the right atrium of the heart and pulmonary artery, leading to life-threatening complications. This case report describes a 43-year-old female patient with IVL who underwent four myomectomy surgeries. Case Report: A 43-year-old female patient with rare IVL was admitted after four myomectomy surgeries. After preoperative evaluation by color Doppler ultrasound, cardiac ultrasound, chest and abdominal computed tomography (CT) scans, this patient underwent total hysterectomy, bilateral salpingectomy, and complete resection of the entire intravascular leiomyoma. This article discusses the characteristics, diagnosis, treatment, and especially surgical precautions of IVL. In addition to surgical resection, this article also discusses other treatment options for IVL. Conclusion: Intravascular leiomyomatosis has unique biological characteristics and clinical manifestations. Through accurate diagnosis and timely complete tumor resection, as well as regular follow-up after surgery, most IVL patients can achieve good long-term prognosis and quality of life. However, further research is needed on the etiology and pathogenesis of IVL.

Abstract A041: Deciphering the bidirectional influences of sympathetic neurons and cancer associated fibroblasts and the resulting contributions to PDAC progression

Abstract The pancreatic ductal adenocarcinoma (PDAC) tumor microenvironment (TME) is innervated by a variety of peripheral nerves that are emerging as facilitators of tumor initiation, progression and metastasis. However, the influences of sympathetic nerves on the TME and how these bi-directional interactions can exacerbate PDAC progression remains unclear. Sympathetic nerves innervate both human and murine PDAC tumors and are frequently surrounded by abundant and heterogeneous cancer-associated fibroblasts (CAFs), which are known to play various crucial roles in PDAC progression. In our studies, we aim to elucidate the bidirectional CAF and sympathetic-neuron crosstalk that may promote tumor growth using a variety of in vitro coculture systems to model sympathetic-CAF interactions, as well as in vivo modeling of nerves in murine PDAC tumors. Our nerve-CAF cocultures reveal that CAFs closely and frequently interact with sympathetic nerves, and also enhance sympathetic nerve outgrowth. We further sequenced these cocultured conditions to evaluate intrinsic transcriptional changes that may be contributing to the enhanced nerve outgrowth. We identified several genes that may contribute to upregulated CAF adhesion, ECM deposition and neuroactive ligand-receptor interactions, which may also be supportive of the axon outgrowth and tumor cell growth. We are also currently characterizing human and murine PDAC samples that contain and lack sympathetic innervation to evaluate spatial phenotypic shifts within the TME. Collectively, our results suggest that this crosstalk between sympathetic neurons and CAFs may both enhance tumor innervation and tumor-promoting phenotypes in the TME, such as axonogenesis and fibrosis in PDAC. Citation Format: Ariana Sattler, Parham Diba, Tetiana Korzun, Marigold Kuykendall, Kevin MacPherson-Hawthorne, Mara Sherman, Teresa Zimmers, Sebnem E Eksi. Deciphering the bidirectional influences of sympathetic neurons and cancer associated fibroblasts and the resulting contributions to PDAC progression [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A041.

Abstract B045: Sensitizing the PDAC tumor microenvironment to immune checkpoint therapies: characterization of a PDAC 3D model to decipher immune infiltration

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal solid tumor with an unfavorable prognosis, often resistant to conventional treatments due to its dense stroma rich in Cancer- Associated Fibroblasts (CAFs). Limited T cell infiltration and prevalence of immunosuppressive cells within the tumor hinder the efficacy of immune checkpoint inhibitors (ICIs). The objective of our project is to modify immune landscape of PDAC tumor microenvironment (TME) to induce ICIs sensitization. We thus develop novel therapeutic strategies combining an immunomodulatory cytokine Interleukine-15 (IL15), with conventional chemotherapies (Gemcitabine or Folfirinox). To accurately replicate the complex cancer-stroma interactions within the pancreatic tumor microenvironment, we constructed a three-dimensional in vitro heterospheroid model composed of xenograft-derived tumor cells from PDAC patients, both primary or immortalized CAFs, and peripheral blood mononuclear cells (PBMCs) from healthy donors. Human 3D models were first set up and characterized by cytometry, imaging mass spectrometry and immunohistochemistry. We showed that, CAFs produce components of the extracellular matrix, promote tumor cell growth, improve resistance to chemotherapy and are able to down-regulate immune cell infiltration and modulate the nature of infiltrated immune cells. Spatial analysis of spheroids using imaging mass cytometry show that CAF influences the distribution of immune cells in our models. Interestingly, Interleukine 15 treatment prompted robust infiltration of PBMCs into heterospheroids. Immunophenotyping experiments revealed a substantial shift in the nature of immune infiltration, marked by a pronounced increase in CD4+ and CD8+ T lymphocytes, gDelta T cell and NK cell populations. When combined with Gemcitabine and IL15, immune effector cell infiltration is remarkably increased, resulting in antitumoral effects and control of tumor cell growth. Moreover, therapeutic combinations induce activation of infiltrated immune effector cells with an increase of activation, degranulation and cytotoxic markers, such as IFNγ, CD107a, and Granzyme B respectively. Thus, the heterotypic spheroids described in our study are a suitable in vitro model to both characterize the influence of CAF on therapeutic effects and the mechanisms that drives immune suppressive microenvironment. Moving forward, the next steps will consist to conduct in vivo experiments using a syngeneic PDAC orthotopic model to assess whether this combined therapeutic approach sensitize the PDAC microenvironment to anti-PD1 therapies. Citation Format: Thomas Bessede, Clara Freixinos, Jennifer Barrat, Véronique Garambois, Nadia Vie, Henri-Alexandre Michaud, Julien Faget, Nathalie Bonnefoy, Céline Gongora, Bruno Robert, Laurent Gros, Christel Larbouret. Sensitizing the PDAC tumor microenvironment to immune checkpoint therapies: characterization of a PDAC 3D model to decipher immune infiltration [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B045.

Abstract B046: Sensitizing the PDAC tumor microenvironment to immune checkpoint therapies: characterization of a PDAC 3D model to decipher immune infiltration

Abstract Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal solid tumor with an unfavorable prognosis, often resistant to conventional treatments due to its dense stroma rich in Cancer- Associated Fibroblasts (CAFs). Limited T cell infiltration and prevalence of immunosuppressive cells within the tumor hinder the efficacy of immune checkpoint inhibitors (ICIs). The objective of our project is to modify immune landscape of PDAC tumor microenvironment (TME) to induce ICIs sensitization. We thus develop novel therapeutic strategies combining an immunomodulatory cytokine Interleukine-15 (IL15), with conventional chemotherapies (Gemcitabine or Folfirinox). To accurately replicate the complex cancer-stroma interactions within the pancreatic tumor microenvironment, we constructed a three-dimensional in vitro heterospheroid model composed of xenograft-derived tumor cells from PDAC patients, both primary or immortalized CAFs, and peripheral blood mononuclear cells (PBMCs) from healthy donors. Human 3D models were first set up and characterized by cytometry, imaging mass spectrometry and immunohistochemistry. We showed that, CAFs produce components of the extracellular matrix, promote tumor cell growth, improve resistance to chemotherapy and are able to down-regulate immune cell infiltration and modulate the nature of infiltrated immune cells. Spatial analysis of spheroids using imaging mass cytometry show that CAF influences the distribution of immune cells in our models. Interestingly, Interleukine 15 treatment prompted robust infiltration of PBMCs into heterospheroids. Immunophenotyping experiments revealed a substantial shift in the nature of immune infiltration, marked by a pronounced increase in CD4+ and CD8+ T lymphocytes, gDelta T cell and NK cell populations. When combined with Gemcitabine and IL15, immune effector cell infiltration is remarkably increased, resulting in antitumoral effects and control of tumor cell growth. Moreover, therapeutic combinations induce activation of infiltrated immune effector cells with an increase of activation, degranulation and cytotoxic markers, such as IFNγ, CD107a, and Granzyme B respectively. Thus, the heterotypic spheroids described in our study are a suitable in vitro model to both characterize the influence of CAF on therapeutic effects and the mechanisms that drives immune suppressive microenvironment. Moving forward, the next steps will consist to conduct in vivo experiments using a syngeneic PDAC orthotopic model to assess whether this combined therapeutic approach sensitize the PDAC microenvironment to anti-PD1 therapies. Citation Format: Thomas Bessede, Clara Freixinos, Jennifer Barrat, Véronique Garambois, Nadia Vie, Henri-Alexandre Michaud, Julien Faget, Nathalie Bonnefoy, Céline Gongora, Bruno Robert, Laurent Gros, Christel Larbouret. Sensitizing the PDAC tumor microenvironment to immune checkpoint therapies: characterization of a PDAC 3D model to decipher immune infiltration [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B046.

Abstract A010: A patient first approach using CRISPR-directed gene editing as an augmentative therapy for the treatment of pancreatic ductile adenocarcinoma

Abstract At the Gene Editing Institute, we are advancing a novel strategy for the treatment of pancreatic ductile adenocarcinoma by employing CRISPR-directed gene editing to disable the functions of key genes that block effective drug application including chemotherapy, radiation and immunotherapy. No matter how careful one is at designing a specific drug against a specific protein, ultimately, the tumor cell develops a workaround. A paradigm shift is required to truly overcome this formidable barrier. We’ve accomplished this by leveraging the effectiveness of standard care and augmenting it, thereby reducing patient suffering, increasing treatment tolerance and, most importantly, improving outcomes. With an enthusiastic response for this concept from the FDA- INTERFACT meeting, we have demonstrated that a carefully selected CRISPR/Cas complex disables the NRF2 gene in the tumor cell and significantly reduces the dose of chemotherapy required to halt pancreatic tumor cell growth, and consequently promoting cell death. The efficiency of gene editing approaches 70 % routinely, which destroys the function of NRF2 and enables additional killing of residual tumor cells by chemotherapeutic agents. As a consequence of CRISPR/Cas activity, we effectively reduce the amount of chemotherapy needed to kill tumor cells by over 20-fold. This level of reduction should enable patients to complete their treatment regimens more successfully, at lower dosages and with far fewer adverse side effects. In addition to reduced suffering, this will invariably prolong survival as we are augmenting already proven treatments. This approach is agnostic to the haplotype of the tumor cell and does not rely on specific biomarker composition; radically expanding the patient population that would be suitable for this type of treatment. We will discuss the impact of CRISPR/Cas-gene editing to disable specific genes, including NRF2 and mutant KRAS, to enable current therapies to work at greater efficacy and at lower dosages. Citation Format: London P McGill, Kelly H Banas, Gregory Tiesi, Eric B Kmiec. A patient first approach using CRISPR-directed gene editing as an augmentative therapy for the treatment of pancreatic ductile adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A010.