RASSF1C modulates expression of PIWI-(Drosophila)-like 1 (PIWIL1), as well as novel PIWI-interacting RNAs (piRNAs), which are regulators of the stem cell phenotype linked to development of lung oncogenesis, in part, through modulation of tumor microenvironment. We reported in previous work that restoring piR-46545 inhibits colony formation and migration/invasion of lung cancer cells overexpressing RASSF1C or PIWIL1 suggesting that it could be an important tumor suppressor. In this article, we identified NEIL2 (nei like DNA glycosylase 2; a base excision repair enzyme) as a direct piR-46545 target gene that linked to promoting lung cancer stem cell (LCSC) drug resistance . We found that over-expression of RASSF1C elevated NEIL2 mRNA levels suggesting that RASF1C could impact LCSC drug resistance. We also report that treatment of lung cancer cells with piR-46545, unlike RASSF1C and PIWIL1, reduces PLOD2 and Vimentin gene expression, key genes associated with tumor microenvironment. In addition, piR-46545 and the anti-cancer drug betulinic acid (BA) appear to display an inhibitory synergistic effect on lung cancer cell proliferation. Based on these findings, we hypothesize that piR-46545 is a potential inhibitor of lung cancer cell growth and progression and it could be a clinically relevant biomarker for lung cancer diagnosis, prognosis, or treatment.

Abstract Introduction: It is important to discover and characterize new pathways and their associated gene networks in cancer cells to develop effective diagnostic and therapeutic tools. Our laboratory is focused on the impact of the RASSF1C-PIWIL1-piRNA pathway on non-small cell lung cancer (NSCLC) cell growth; and our studies have resulted in identifying specific PIWI-interacting RNAs (piRNAs) that appear to function both as oncogenes and tumor suppressors. Among the recent piRNAs identified, we found that piR-60643 targets fatty acid bind protein 5 (FABP5) in lung cancer and appears to function as a tumor suppressor. Methods: In previously published work we reported on a global microarray screen which identified potential piRNA genes in non-small lung cancer cells over-expressing RASSF1C. We were able to detect several piRNAs that are modulated by RASSF1C. The expression of specific piRNAs that appear to be down or up-regulated by RASSF1C were confirmed by RT-PCR. We evaluated the impact of selected RASSF1C-target piRNAs on lung cancer cell proliferation and migration in vitro. We also have identified both host and target genes for specific piRNAs and have used piRNA mimics to down regulate target gene expression. Results: We show that piR-60643 is down regulated by RASSF1C and seems to function as a tumor suppressor in non-small cell lung cancer cells. Consistent with this, treatment of lung cancer cells with piR-60643 mimics reduced cell proliferation and migration, supporting the idea that piR-60643 could be a new inhibitor of lung cancer cell growth and progression. To learn more about how piR-60643 may hinder lung cancer cell growth, we searched for genes that could be targeted by piR-60643. Interestingly, we identified FABP5 as a piR-60643-target gene. FABP5 is up regulated in lung tumor tissue and in tissues of other cancers and is associated with poor patient survival. FABP5 plays a key role in regulating lipid metabolism and fatty acid oxidation (FAO). FAO is critical to supply the high energy (ATP) demand of rapidly growing cancer cells and contributes to cell cycle progression, EMT, migration, and metastasis. We found that treatment of lung cancer cells with piR-60643 mimics resulted in down regulation of FABP5 expression both at the mRNA and protein levels, implying that piR-60643 could disrupt FAO-associated metabolic pathway(s) in lung cancer cells. Interestingly, we found that PIWIL1 overexpression up-regulates FABP5 expression in lung cancer cells, suggesting that the PIWIL-FABP5 gene axis may play a role in lipid metabolism/FAO. This novel finding has not been previously reported. Conclusion: Targeting FABP5 by piR-60643 could prove a novel mechanism of inhibiting lipid metabolism/FAO and impeding tumor growth and progression. As such, piR-60643 could prove to be a useful diagnostic/therapeutic tool for lung cancer. Citation Format: Yousef G. Amaar, Mark E. Reeves. PiR-60643 is a novel inhibitor of fatty acid binding protein 5 (FABP5) in lung cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2067.

Pancreatic ductal adenocarcinoma (PDAC) is a devastating malignancy with a 5-year overall survival (OS) rate of approximately 12

Outcomes 1. Learners will gain insight into how Electronic Patient Reported Outcomes (ePROs) can provide objective assessments of patient symptoms and quality of life, enhancing clinical management and treatment response evaluation.2. Learners will be familiar with the step-by-step implementation process of ePROs, including stakeholder involvement, tool selection, and integration into electronic health records (EHR). Key Message Electronic Patient Reported Outcomes (ePROs) are crucial indicators to develop a palliative care clinic. Implementing ePROs is challenging, as there are many barriers in an increasingly complex health care system. Here, we describe the process for successful implementation of ePROs in a Palliative Care clinic and a Comprehensive Cancer Center. Abstract Introduction Electronic Patient Reported Outcomes (ePROs) measure symptom assessment objectively, providing raw insights into a patient's quality of life. ePROs enhance objectivity for clinical management identifying unmet needs, informing disease progression and treatment response. We describe the implementation process of ePROs at the Loma Linda University Cancer Center. Methods ePROs implementation began in August 2020, under Palliative Care (PC) leadership, involving key stakeholders such as clinic staff, physicians and hospital leadership. We chose the Edmonton Symptom Assessment Scale-Financial and Spiritual scale (ESAS-FS) because of its simplicity and reproducibility (1). We collected responses from patients using ESAS-FS before or during a clinic visit. A flowchart embedded ePROs in our EHR. We also created a Tableau automated report with demographic information, while medication use, and hospital utilization was manually abstracted. ePROs were piloted in a single Palliative Medicine clinic, then expanded to all 4 clinics. Later, we piloted it in the Cancer Center. Results We analyzed data on patient demographics and ESAS-FS entries from November 2020 to June 2024. 1259 patients completed 4297 ESAS-FS entries. The cohort comprised 53.6% female and 46.4% male patients, with a racial distribution of 66.7% White, 13.4% Black, 7.7% Hispanic, 6.4% Asian, and 5.5% other. ESAS-FS submissions increased annually, with a 15.6% rise from 2022 to 2023. Additionally, manually extracted data from 109 PC clinic patients from January 2021 to December 2023 revealed a decrease in average ESAS-FS scores from 39.9 at the initial visit to 35.6 on their last visit. Conclusion Our experience implementing PROs within the Cancer Center demonstrates the feasibility and scalability of our project for any EHR system. We propose a method of implementation, ongoing evaluation, tool refinement, and automated reporting. Our work shows the importance of implementation beyond the PC clinic to the Cancer Center. References 1. Hui, David, and Eduardo Bruera . “The Edmonton Symptom Assessment System 25 Years Later: Past, Present, and Future Developments.” Journal of Pain and Symptom Management, Elsevier, 29 Dec. 2016, www.sciencedirect.com/science/article/pii/S0885392416312131.

This case-control study evaluates whether adjuvant radiotherapy is associated with overall survival among patients with surgically resected stage III Merkel cell carcinoma.

Acute myeloid leukemia (AML) has a poor survival rate for both pediatric and adult patients due to its frequent relapse. To elucidate the bioenergetic principle underlying AML relapse, we investigated the transcriptional regulation of mitochondrial–nuclear dual genomes responsible for metabolic plasticity in treatment-resistant blasts. Both the gain and loss of function results demonstrated that NFκB2, a noncanonical transcription factor (TF) of the NFκB (nuclear factor kappa-light-chain-enhancer of activated B cells) family, can control the expression of TFAM (mitochondrial transcription factor A), which is known to be essential for metabolic biogenesis. Furthermore, genetic tracking and promoter assays revealed that NFκB2 is in the mitochondria and can bind the specific “TTGGGGGGTG” region of the regulatory D-loop domain to activate the light-strand promoter (LSP) and heavy-strand promoter 1 (HSP1), promoters of the mitochondrial genome. Based on our discovery of NFκB2′s novel function of regulating mitochondrial–nuclear dual genomes, we explored a novel triplet therapy including inhibitors of NFκB2, tyrosine kinase, and mitochondrial ATP synthase that effectively eliminated primary AML blasts with mutations of the FMS-related receptor tyrosine kinase 3 (FLT3) and displayed minimum toxicity to control cells ex vivo. As such, effective treatments for AML must include strong inhibitory actions on the dual genomes mediating metabolic plasticity to improve leukemia prognosis.

Background: T-cell-based adoptive cell therapies have emerged at the forefront of cancer immunotherapies; however, failed long-term survival and inevitable exhaustion of transplanted T lymphocytes in vivo limits clinical efficacy. Leukemia blasts possess enhanced glycolysis (Warburg effect), exploiting their microenvironment to deprive nutrients (e.g., glucose) from T cells, leading to T-cell dysfunction and leukemia progression. Methods: Thus, we explored whether genetic reprogramming of T-cell metabolism could improve their survival and empower T cells with a competitive glucose-uptake advantage against blasts and inhibit their uncontrolled proliferation. Results: Here, we discovered that high-glucose concentration reduced the T-cell expression of glucose transporter GLUT1 (SLC2A1) and TFAM (mitochondrion transcription factor A), an essential transcriptional regulator of mitochondrial biogenesis, leading to their impaired expansion ex vivo. To overcome the glucose-induced genetic deficiency in metabolism, we engineered T cells with lentiviral overexpression of SLC2A1 and/or TFAM transgene. Multi-omics analyses revealed that metabolic reprogramming promoted T-cell proliferation by increasing IL-2 release and reducing exhaustion. Moreover, the engineered T cells competitively deprived glucose from allogenic blasts and lessened leukemia burden in vitro. Conclusions: Our findings propose a novel T-cell immunotherapy that utilizes a dual strategy of starving blasts and cytotoxicity for preventing uncontrolled leukemia proliferation.

Acute myeloid leukemia (AML) is a hematological malignancy that is characterized by an expansion of immature myeloid precursors. Despite therapeutic advances, the prognosis of AML patients remains poor and there is a need for the evaluation of promising therapeutic candidates to treat the disease. The objective of this study was to evaluate the efficacy of duocarmycin Stable A (DSA) in AML cells in vitro. We hypothesized that DSA would induce DNA damage in the form of DNA double-strand breaks (DSBs) and exert cytotoxic effects on AML cells within the picomolar range. Human AML cell lines Molm-14 and HL-60 were used to perform 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), DNA DSBs, cell cycle, 5-ethynyl-2-deoxyuridine (EdU), colony formation unit (CFU), Annexin V, RNA sequencing and other assays described in this study. Our results showed that DSA induced DNA DSBs, induced cell cycle arrest at the G2M phase, reduced proliferation and increased apoptosis in AML cells. Additionally, RNA sequencing results showed that DSA regulates genes that are associated with cellular processes such as DNA repair, G2M checkpoint and apoptosis. These results suggest that DSA is efficacious in AML cells and is therefore a promising potential therapeutic candidate that can be further evaluated for the treatment of AML.

Studies have demonstrated comparable outcomes between laparoscopic and open resection of gastrointestinal stromal tumor (GIST). We sought to compare outcomes among robotic, laparoscopic, and open resection of gastric GIST in the era of expanding minimally invasive surgery. A retrospective analysis was performed of adult patients with gastric GIST undergoing definitive surgery using the National Cancer Database from 2010 to 2020, excluding cases converted to open. Patients were stratified into minimally invasive surgery (MIS), (combined robotic (R) and laparoscopic (L)), and open (O). Hospital length of stay (LOS), 30-day mortality, 90-day mortality, and margin status were assessed. Subgroup analysis was performed to evaluate outcomes between R and L cohorts. Entropy balancing was used to adjust for intergroup differences. Kaplan–Meier survival estimates were used to compare unadjusted 5-year survival. Of the 15,022 patients (R = 10.4

Abstract Introduction: It is crucial to identify new driver genes and associated downstream pathways to facilitate the development of targeted therapies to effectively treat late-stage and metastatic lung cancer. One pathway involves the Ras association domain family 1 (RASSF1) gene which encodes multiple isoforms. Our laboratory has discovered that the RASSF1C isoform exhibits functional characteristics of an oncogene. We discovered that RASSF1C modulates the expression of PIWI-(Drosophila)-like 1 (PIWIL1, a stem cell renewal gene), as well as novel PIWI-interacting RNAs (piRNAs), which are regulators of the stem cell phenotype and have been linked to oncogenesis. While studying this pathway, we have identified novel piRNAs that are modulated by RASSF1C in non-small cell lung cancer (NSCLC) cells that appear to act as oncogenes or tumor suppressors. Methods: We conducted a global microarray screen to identify RASSF1C PIWI-interacting RNA (piRNA) gene targets. Several piRNA genes that are up- and down-regulated by RASSF1C were identified and the expression of selected up-regulated and down-regulated target genes were confirmed by RT-PCR. We evaluated the impact of selected piRNAs on lung cancer cell proliferation and migration in vitro. We also assessed the expression of the validated piRNA target genes in lung tumor and matched normal tissues. Host and target genes for specific piRNAs were identified using a piRNA database (piRNAdb.org). Results: Among the down-regulated piRNAs, we found that restoring piR-50485 expression drastically decreased lung cancer cell proliferation and invasion, suggesting that piR-50485 could be a potent inhibitor of lung cancer cell growth and progression. Consistent with this, we found that piR-50485 expression is significantly down-regulated in about 60% of human lung cancers. We also identified that the ZHX3 gene (a transcription repressor), which is down-regulated in human lung cancers, is the host gene for piR-50485. In addition, we identified the ZNF618 gene (a transcription co-regulator) that is up-regulated in lung tumor tissue, is a piR-50485 target gene. Consistent with this, treatment of lung cancer cells with piR-50485 mimics resulted in a significant decrease of ZNF618 protein levels, cell proliferation, and migration. Further, increased expression levels of ZHX3 are associated with higher lung cancer patient survival, while increased ZNF618 levels are associated with lower survival. Our novel findings suggest the hypothesis that the piR-50485-ZHX3-ZNF618 gene axis could negatively impact lung cancer cell growth and progression. Conclusion: Exploring the impact of the piR-50485-ZHX3-ZNF618 gene axis on lung cancer growth is a novel concept about which nothing is currently known. We are studying piR-50485 to determine if it can be an effective therapy for lung cancer metastasis, and whether it provides important prognostic information about lung cancer outcomes. Citation Format: Mark E. Reeves, Yousef G. Amaar. PIWI-interacting RNA 50485 (piRNA-50485) is a novel lung tumor suppressor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 469.

In the past decade, targeted therapies for solid tumors, including non-small cell lung cancer (NSCLC), have advanced significantly, offering tailored treatment options for patients. However, individuals without targetable mutations pose a clinical challenge, as they may not respond to standard treatments like immune-checkpoint inhibitors (ICIs) and novel targeted therapies. While the mechanism of action of ICIs seems promising, the lack of a robust response limits their widespread use. Although the expression levels of programmed death ligand 1 (PD-L1) on tumor cells are used to predict ICI response, identifying new biomarkers, particularly those associated with the tumor microenvironment (TME), is crucial to address this unmet need. Recently, inflammatory cytokines such as interleukin-1 beta (IL-1β) have emerged as a key area of focus and hold significant potential implications for future clinical practice. Combinatorial approaches of IL-1β inhibitors and ICIs may provide a potential therapeutic modality for NSCLC patients without targetable mutations. Recent advancements in our understanding of the intricate relationship between inflammation and oncogenesis, particularly involving the IL-1β/PD-1/PD-L1 pathway, have shed light on their application in lung cancer development and clinical outcomes of patients. Targeting these pathways in cancers like NSCLC holds immense potential to revolutionize cancer treatment, particularly for patients lacking targetable genetic mutations. However, despite these promising prospects, there remain certain aspects of this pathway that require further investigation, particularly regarding treatment resistance. Therefore, the objective of this review is to delve into the role of IL-1β in NSCLC, its participation in inflammatory pathways, and its intricate crosstalk with the PD-1/PD-L1 pathway. Additionally, we aim to explore the potential of IL-1β as a therapeutic target for NSCLC treatment.

Understanding the factors which shape T-lymphocyte immunity is critical for the development and application of future immunotherapeutic strategies in treating hematological malignancies. The thymus, a specialized central lymphoid organ, plays important roles in generating a diverse T lymphocyte repertoire during the infantile and juvenile stages of humans. However, age-associated thymic involution and diseases or treatment associated injury result in a decline in its continuous role in the maintenance of T cell-mediated anti-tumor/virus immunity. Acute myeloid leukemia (AML) is an aggressive hematologic malignancy that mainly affects older adults, and the disease’s progression is known to consist of an impaired immune surveillance including a reduction in naïve T cell output, a restriction in T cell receptor repertoire, and an increase in frequencies of regulatory T cells. As one of the most successful immunotherapies thus far developed for malignancy, T-cell-based adoptive cell therapies could be essential for the development of a durable effective treatment to eliminate residue leukemic cells (blasts) and prevent AML relapse. Thus, a detailed cellular and molecular landscape of how the adult thymus functions within the context of the AML microenvironment will provide new insights into both the immune-related pathogenesis and the regeneration of a functional immune system against leukemia in AML patients. Herein, we review the available evidence supporting the potential correlation between thymic dysfunction and T-lymphocyte impairment with the ontogeny of AML (II-VI). We then discuss how the thymus could impact current and future therapeutic approaches in AML (VII). Finally, we review various strategies to rejuvenate thymic function to improve the precision and efficacy of cancer immunotherapy (VIII).

The tumor microenvironment (TME) plays a vital role in tumor invasion and metastasis and provides a rich environment for identifying novel therapeutic targets. The TME landscape consists of an extracellular matrix (ECM) and stromal cells. ECM is a major component of TME that mediates the interaction between cancer cells and stromal cells to promote invasion and metastasis. We have shown in published work that RASSF1C promotes cancer stem cell development, migration, and drug resistance, in part, by promoting EMT through a mechanism that involves up-regulation of the PIWIL1-piRNA axis. Consistent with this, in this study, we demonstrate that RASSF1C promotes lung cancer metastasis in vivo using an orthotopic mouse model. Interestingly, two target genes identified in a previously conducted microarray study to be up-regulated by RASSF1C in breast and non-small cell lung cancer (NSCLC) cells are prolyl 4-hydroxylase alpha-2 (P4HA2) and procollagen-lysine, 2-oxoglutarate 5-dioxygenase 2 (PLOD2). In cancer, P4H2A and PLOD2 are vital for collagen posttranslational modification and folding leading to the formation of a stiff ECM and induction of EMT and cancer stem cell marker gene expression, resulting in metastatic dissemination. Here, we also show that overexpression of RASSF1C up-regulates Collagen I, P4HA2, and PLOD2 in vitro. Up-regulation of P4HA2 and PLOD2 by RASSF1C was also confirmed in lung and breast cancer cells in vivo using mouse models. Further, we found that treatment of wildtype lung cancer cells or lung cancer cells overexpressing RASSF1C or PIWIL1 with piR-35127 and 46545 (both down-regulated by RASSF1C) decreased lung cancer cell invasion/migration. Taken together, our findings suggest that RASSF1C may promote lung cancer cell ECM remodeling to induce lung cancer cell stemness, invasion, and metastasis, in part, by up-regulating a previously unknown PIWIL1-P4HA2-PLOD2 pathway. Furthermore, piR-35127 and piR-46545 could potentially be important anti-metastatic tools.

BackgroundThis study compares survival rates, recurrence patterns, toxicity, and treatment cost in patients with hepatocellular carcinoma (HCC) treated with either transarterial chemoembolization (TACE) or proton beam radiotherapy (PBT). MethodsSubjects with untreated HCC meeting Milan or San Francisco transplant criteria were recruited. Subjects were randomized to receive PBT (n = 36) or TACE (n = 40). Proton therapy was administered in 15 fractions over 3 weeks to a total dose of 70.2 Gy. TACE was repeated until complete or maximal response. The primary outcome measure was overall survival (OS). Secondary end points were progression-free survival (PFS), local control (LC), toxicity, and cost. ResultsOf the 76 randomized patients, 74 were assessed for outcome measures. The 2-year OS for PBT versus TACE was similar at 68%, 95% confidence interval (CI), 0.54-0.86, and 65%, 95% CI, 0.52-0.83 (p = .80), however, median PFS was improved for PBT versus TACE (not reached vs. 12 months, p = .002). LC was improved with PBT versus TACE (hazard ratio, 5.64; 95% CI, 1.78-17.9, p = .003). Days of posttreatment hospitalization were 24 for PBT and 166 for TACE (p < .001). Total mean cost per patient for treatment and posttreatment care revealed a 28% cost savings for PBT. ConclusionsPBT and TACE yielded similar OS for treatment of HCC, but PFS and LC were improved with PBT compared to TACE. Patients treated with PBT required fewer courses of treatment, fewer posttreatment hospitalization days, and reduced cost of treatment compared to TACE. These data support the use of PBT as a viable treatment alternative to TACE for patients with HCC within transplant criteria.

In the past decade, targeted therapies for solid tumors, including non-small cell lung cancer (NSCLC), have advanced significantly, offering tailored treatment options for patients. However, individuals without targetable mutations pose a clinical challenge, as they may not respond to standard treatments like immune-checkpoint inhibitors (ICIs) and novel targeted therapies. While the mechanism of action of ICIs seems promising, the lack of a robust response limits their widespread use. Although the expression levels of programmed death ligand 1 (PD-L1) on tumor cells are used to predict ICI response, identifying new biomarkers, particularly those associated with the tumor microenvironment (TME), is crucial to address this unmet need. Recently, inflammatory cytokines such as interleukin-1 beta (IL-1β) have emerged as a key area of focus and hold significant potential implications for future clinical practice. Combinatorial approaches of IL-1β inhibitors and ICIs may provide a potential therapeutic modality for NSCLC patients without targetable mutations. In this review, we discuss the role of IL-1β in NSCLC, its involvement in inflammatory pathways, and explore its potential role in the treatment of NSCLC.