Cell Rescue Research Articles

Outcomes of R-MPV followed by ara-c in primary central nervous system lymphoma: A single-center retrospective analysis.

e19046 Background: Primary central nervous system lymphoma (PCNSL) lymphoma is a rare non-Hodgkin’s Lymphoma. Currently, there is no uniform consensus in the optimal management of PCNSL. In patients that can tolerate systemic therapy, induction therapy is typically done with a high-dose methotrexate-based regimen, due to evidence of better penetration of the blood brain barrier. One such regimens is high dose methotrexate combined with rituximab, procarbazine, and vincristine (R-MPV). Options for consolidation therapy include high-dose systemic therapy with stem cell rescue (ASCT), whole brain radio therapy (WBRT) and high-dose cytarabine (ara-c) with or without etoposide (EA). At our institution we utilize an R-MPV induction approach with some form of ara-c consolidation without etoposide. We aimed to execute a retrospective cohort study to analyze outcomes of this approach. Methods: All the patients diagnosed with PCNSL who received an induction regimen of R-MPV from 10/01/2020 to 06/01/2023 were included. Data was then gathered by chart review of the electronic medical record. Treatment outcomes were evaluated based on imaging with brain MRIs and PET scans. Central findings such as complete response (CR) rate, progression free survival (PFS) and overall survival (OS) were ascertained. Results: 10 charts were reviewed. 9 out of the 10 patients had a histological presentation consistent with diffuse large B-cell lymphoma; 1 patient had a diagnosis of B-cell lymphoma. In terms of demographics, the patients ages ranged from 54 to 83 years, with a median of 74 years. Complete response was achieved in 5 out of the 10 patients (CR rate of 50%), and no relapse was noted in any of these patients. 2 out of the 10 patients progressed on treatment, with 1 dying from progression of the disease. The median follow-up for patients who were still alive was 24 months. 2-year PFS was 80%, and 2-year OS was 90%. Median PFS and OS were not reached. Mean PFS and OS were 28 months and 30.5 months respectively. Conclusions: Based on our study results, the CR rate we observed for R-MPV appears consistent with existing literature on this regimen and other methotrexate-based induction regimens. Our results for R-MPV with ara-c consolidation provides similar outcomes to existing studies on treatment with different methotrexate-based induction and consolidation with ASCT, particularly the 80% PFS we observed. This percent PFS also appears similar to studies of consolidation with ara-c and EA, and superior to PFS with WBRT. Overall, these results are reassuring as in a specific set of older patients who might not be candidates for ASCT, our specific nonmyeloablative approach could be worthwhile. However, our study does have the limitation of being a single center retrospective perspective. [Table: see text]

Marrow-ablative consolidation chemotherapy (HDCT) to decrease the development of choroid plexus carcinoma (CPC) specific relapse in high-risk TP53-mutated (germline or somatic) young children with newly diagnosed CPC.

2088 Background: Choroid plexus carcinomas (CPC) are highly malignant, early childhood cancers in which the loss of TP53 function is a central element of their development and defines unfavorable disease. To date, CPC-free survival rates have remained poor; recent data from St. Jude including complete TP53 mutation status, yielded CPC-free survival of only 29% with a non-marrow-ablative chemotherapy strategy. The rarity of CPC has constrained the ability to explore the role of HDCT with autologous hematopoietic progenitor cell rescue (AuHPCR). Previously reported results of the "Head Start" (HS) HDCT approach showed some promise of cure, but were limited by small patient numbers, short follow-up and incomplete TP53 mutation data. Consequently, the role of HDCT and AuHPCR in TP53-mutated CPC remains unclear. Methods: We have analyzed data on TP53 status, therapy, treatment responses, CPC-free and overall survivals (OS) and second cancers from the largest cohort of such patients, all who received initial therapy with the intent/consideration of undergoing consolidative HDCT. Results: Twenty-seven children with CPC (median age, 16 months) received HS-like induction chemotherapy regimens or ICE with the intent of utilizing consolidative HDCT. Twenty-three of 27 completed HDCT consolidation; the remaining 4 patients did not proceed with HDCT due to parental choice. Somatic or germline TP53 mutations were identified in 17 patients, while 4 patients exhibited TP53 wild type (TP53wt); TP53 status remains untested or unavailable in 6 patients. Five-year CPC-free survival and OS for the entire cohort were 47.8% and 44.4%. CPC-free survival for those with TP53 germline mutations (n=12) was 66%, for those with somatic tumor TP53 mutations (n=5) was 40% and for those with confirmed TP53wt (n=4) was 50%. The 5-year CPC-free survival for all 23 patients who underwent HDCT and AuHPCR was 52%, comprising 3/4 TP53wt patients, 10/12 TP53 germline mutated patients and all 5 TP53 somatic tumor mutated patients. Notably, all six survivors with TP53 mutations had undergone HDCT with AuHPCR, and all but one avoided irradiation. Eight out of 12 patients with documented genotypic (TP53 germline mutated) Li-Fraumeni Cancer Predisposition Syndrome (LFS) developed second malignancies. Conclusions: Our data indicate that HDCT with AuHPCR consolidation is associated with improved CPC-free survival compared with historical reports in young children with high-risk TP53 mutated CPC. However, the ultimate development of second cancers substantially affects outcomes for those with LFS, irrespective of HDCT treatment. These data justify the inclusion of HDCT and AuHPCR in the prospective international trial under development for young children with newly diagnosed TP53 mutated CPC.

Genome-wide analysis of polyphenol oxidase gene family in Solanum tuberosum and functional identification of StuPPO9 in regulating tolerance to Phytophthora infestans

Polyphenol oxidase (PPO) is the key enzyme involved in the enzymatic browning of potato tubers. However, the understanding of its other functions, as well as the underlying molecular mechanisms, is still limited. StuPPO9 was predicted as an inducible gene in response to disease defense and cell rescue, as analyzed using the EST database. In this study, an identification and evolutionary analysis of PPOs in Solanum tuberosum was conducted. A total of 10 members of StuPPOs were identified and their gene structure, protein motifs, chromosomal location, and tissue distribution were further investigated. QRT-PCR analysis indicated that most PPOs were induced or inhibited under hormone treatments, and StuPPO9 showed the highest expression level when induced by Phytophthora infestans. Therefore, StuPPO9, the only one located on chromosome 2, was selected for further study. The StuPPO9 overexpressing Nicotiana benthamiana transgenic line effectively slowed down the growth of P. infestans and showed more tolerance compared to the wild type (WT). Moreover, the molecular markers NtHin1 and PR genes were significantly upregulated in the transgenic tobacco than in WT under P. infestans infection. This study will provide a basis and reference for future research on the roles of StuPPO9 in disease defense.

FDA Approval Summary: Ciltacabtagene Autoleucel for Relapsed or Refractory Multiple Myeloma.

In February 2022, the FDA approved ciltacabtagene autoleucel, a chimeric antigen receptor (CAR) T-cell therapy targeting the B-cell maturation antigen, for adult patients with relapsed/refractory multiple myeloma after ≥4 lines of therapy, including an immunomodulatory agent, a proteasome inhibitor, and an anti-CD38 monoclonal antibody. Approval was based on overall response rate (ORR), complete response (CR) rate, and duration of response (DoR) in 97 adult patients in a single-arm, open-label, multicenter phase 2 trial (CARTITUDE-1 [NCT03548207]). Patients received a single infusion of ciltacabtagene autoleucel, preceded by lymphodepleting chemotherapy. Of the 97 patients evaluable, ORR was 97.9% [95% confidence interval (CI), 92.7-99.7] with a stringent CR rate of 78.4% (95% CI, 68.8-86.1). After median follow-up of 18 months, the median DoR was 21.8 months (95% CI, 21.8-not estimable [NE]) in responders (PR or better) and NE (95% CI, 21.8 months-NE) in patients who achieved stringent CR. Serious adverse reactions occurred in 55% of the 97 patients evaluated for safety. Grade 3 or higher cytokine release syndrome (CRS) and neurologic toxicities occurred in 5% and 11% of the patients, respectively, leading to a Risk Evaluation and Mitigation Strategy. Neurologic toxicities included immune effector cell-associated neurologic syndrome, typically seen with CAR-T products, parkinsonism, peripheral neuropathy, cranial nerve palsies, and Guillain-Barré syndrome. One fatal case of hemophagocytic lymphohistiocytosis/macrophage activation syndrome occurred. Prolonged and recurrent grade 3 or 4 cytopenias occurred; a single patient required hematopoietic stem-cell rescue.

Hypoxia and Giardia duodenalis: The Role of Hypoxia-Inducible Factor in Altered Epithelial Glucose Metabolism

The gastrointestinal system experiences frequent oxygenic fluctuations and must be able to maintain barrier integrity during periods of suboptimal oxygen concentration, or hypoxia. Epithelial cells rely on the Hypoxia-Inducible Factor (HIF) complex to activate genes that combat cellular stress and adapt cellular metabolism during hypoxia. Importantly, protozoan parasites can modulate host tissue oxygen tension and HIF activation, yet little is known regarding the relationship between enteric protozoa and hypoxia. This research aims to uncover the role of HIF upon Giardia duodenalis infection, a top cause of global diarrheal disease and an excellent infection model for the study of GI physiology. We hypothesize that HIF-target genes are activated upon Giardia infection, promoting alternative cellular glucose metabolism to sustain bioenergetic homeostasis. Caco-2 colonic epithelial cells were infected with Giardia isolate GS/M (MOI 10) for 1.5 or 4.5 hours under normoxic (21% O2) or hypoxic (~1%O2, STEMCELL hypoxia incubator) conditions to capture the early or peak activation of the oxygen-dependent HIF subunit (HIF-1α). RNA was extracted for assessment of transcriptional alterations of HIF-target genes via reverse transcriptase quantitative polymerase chain reaction. Investigation of HIF-mediated intracellular metabolic changes was carried out via liquid-chromatography mass-spectrometry analysis (hydrophilic-interaction chromatography method) on cocultures supplemented with a hypoxia mimetic (DMOG) or a HIF inhibitor (PX-478). Metabolomics experiments were repeated using Caco-2 cells transfected with sodium-dependent glucose cotransporter 1 (SGLT1), a key glucose transporter in the small intestine where Giardia localizes which is not reliably expressed in Caco-2 cells. Under normoxic conditions, genes that aid in cell stress responses ( VEGFA, ANKRD37, GADD45A) and glycolysis ( HK2, LDHA) are upregulated in Giardia-infected cells in a time-dependent manner (p<0.05). Fewer HIF-target genes are upregulated under hypoxic conditions (e.g., VEGFA, GADD45A, PGK1; p<0.05), indicating Giardia-infected cells exhibit a transcriptional profile similar to hypoxic uninfected cells. Interestingly, HIF1α was upregulated in Giardia-infected cells under hypoxic conditions at 1.5 hours (p<0.05). Analysis of the Caco-2 intracellular metabolome indicated HIF-dependent changes to nucleic acid (e.g., guanosine, inosine, uracil) and amino acid (e.g., glycine, threonine) metabolism. DMOG treatment increased the abundance of glycolytic intermediates (e.g., PEP, DHAP) in both cell lines and inhibited the depletion of the Kreb’s Cycle intermediate aconitate in the Caco-2-SGLT1 transfected cells, confirming the promotion of glycolytic flux by HIF. Taken together, our findings indicate Giardia-infected cells illustrate a hypoxic signature, a novel metabolic cell rescue mechanism in response to enteropathogens. Elucidating the role of HIF during enteric parasitic infections will aid in our understanding of the cellular adaptations underpinning GI pathophysiology, while also shedding light on the potential of HIF as a therapeutic target. Natural Science and Engineering Research Council of Canada. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Myeloablative vs nonmyeloablative consolidation for primary central nervous system lymphoma: results of Alliance 51101

AbstractAlthough it is evident that standard-dose whole-brain radiotherapy as consolidation is associated with significant neurotoxicity, the optimal consolidative strategy for primary central nervous system lymphoma (PCNSL) is not defined. We performed a randomized phase 2 clinical trial via the US Alliance cancer cooperative group to compare myeloablative consolidation supported by autologous stem cell transplantation with nonmyeloablative consolidation after induction therapy for PCNSL. To our knowledge, this is the first randomized trial to be initiated that eliminates whole-brain radiotherapy as a consolidative approach in newly diagnosed PCNSL. Patients aged 18 to 75 years were randomly assigned in a 1:1 manner to induction therapy (methotrexate, temozolomide, rituximab, and cytarabine) followed by consolidation with either thiotepa plus carmustine and autologous stem cell rescue vs induction followed by nonmyeloablative, infusional etoposide plus cytarabine. The primary end point was progression-free survival (PFS). A total of 113 patients were randomized, and 108 (54 in each arm) were evaluable. More patients in the nonmyeloablative arm experienced progressive disease or death during induction (28% vs 11%; P = .05). Thirty-six patients received autologous stem cell transplant, and 34 received nonmyeloablative consolidation. The estimated 2-year PFS was higher in the myeloablative vs nonmyeloablative arm (73% vs 51%; P = .02). However, a planned secondary analysis, landmarked at start of the consolidation, revealed that the estimated 2-year PFS in those who completed consolidation therapy was not significantly different between the arms (86% vs 71%; P = .21). Both consolidative strategies yielded encouraging efficacy and similar toxicity profiles. This trial was registered at www.clininicals.gov as #NCT01511562.

Successful treatment of paediatric refractory Hodgkin lymphoma with immunotherapy - A case report and literature review.

To describe a rare case of primary refractory Hodgkin lymphoma nodular sclerosis syncytial variant in a child and review immunotherapy in relapsed/refractory Hodgkin lymphoma. We described the treatment course of a child with primary refractory classic Hodgkin lymphoma and discussed different options for salvage therapy, with an emphasis on immunotherapy. We searched PubMed for all published clinical trials investigating immunotherapy in classic Hodgkin lymphoma written in English until 31 June, 2023. The reference list of each identified paper was searched for additional publications. Our patient was salvaged with anti-programmed cell death 1 (PD-1) antibody therapy followed by high-dose chemotherapy with autologous stem cell rescue. Radiotherapy was avoided. We identified five one-armed phase II trials investigating anti-PD-1 therapy in first relapse/refractory disease in a total of 254 patients aged 9-71 years, of which one included 31 children. The complete remission rate before high-dose chemotherapy was 59%-95% overall and 67%-89% among those with refractory disease. Although it remains to be proven in randomised trials, anti-PD-1 therapy may provide higher complete response rates than traditional chemotherapy. Anti-PD-1 therapy has the potential to increase the chance of cure while decreasing the risk of late effects from chemotherapy and radiotherapy.

Autologous HSCT with novel agent-based induction and consolidation followed by lenalidomide maintenance for untreated multiple myeloma.

Triplet regimen comprising proteasome inhibitors, immunomodulatory drugs, and dexamethasone (DEX) is a recommended induction/consolidation therapy for multiple myeloma (MM) patients eligible for transplant. In this Japanese phase II study conducted from 2017 to 2019, newly diagnosed MM patients aged 20-65 received four induction cycles with bortezomib (Bor), lenalidomide (Len), and DEX (VRD), followed by Bor and high-dose melphalan with autologous stem cell rescue. Subsequently, they underwent four consolidation cycles with carfilzomib, Len, and DEX (KRD), followed by Len maintenance until disease progression. A total of 141 patients were analyzed. In an intent-to-treat population, the complete or better response post induction was 19.9%, rising to 39.7%, 58.9%, and 62.4% after transplant, consolidation, and 1-year maintenance, respectively. With a median follow-up of 38 months, the 3-year progression-free survival (PFS) rate was 83.5% and the 3-year overall survival rate was 92.5%. Severe adverse events (≥grade 3) occurred in ~30% of patients; however, there was no treatment-related mortality. These findings clearly showed the tolerability and effectiveness of this protocol. Nevertheless, patients with high-risk cytogenetics showed a trend toward lower 3-year PFS than those without (77.8% vs. 89.4%, p = 0.051), and ultra-high-risk cytogenetics (≥2 high-risk cytogenetics) had an even worse prognosis, with 61.2% 3-year PFS. To overcome this situation, a more potent treatment strategy incorporating novel agents such as the CD38-antibody should be assessed in future studies.

Minimum intensity projection of embossed quadrant-detection images for improved photoreceptor mosaic visualisation.

Non-confocal split-detection imaging reveals the cone photoreceptor inner segment mosaic in a plethora of retinal conditions, with the potential of providing insight to ageing, disease, and response to treatment processes, in vivo, and allows the screening of candidates for cell rescue therapies. This imaging modality complements confocal reflectance adaptive optics scanning light ophthalmoscopy, which relies on the waveguiding properties of cones, as well as their orientation toward the pupil. Split-detection contrast, however, is directional, with each cone inner segment appearing as opposite dark and bright semicircles, presenting a challenge for either manual or automated cell identification. Quadrant-detection imaging, an evolution of split detection, could be used to generate images without directional dependence. Here, we demonstrate how the embossed-filtered quadrant-detection images, originally proposed by Migacz etal. for visualising hyalocytes, can also be used to generate photoreceptor mosaic images with better and non-directional contrast for improved visualisation. As a surrogate of visualisation improvement between legacy split-detection images and the images resulting from the method described herein, we provide preliminary results of simple image processing routines that may enable the automated identification of generic image features, as opposed to complex algorithms developed specifically for photoreceptor identification, in pathological retinas.

A40 THE ROLE OF HYPOXIA-INDUCIBLE FACTOR IN CELLULAR METABOLIC ADAPTATION UPON GIARDIA INFECTION

Abstract Background Recent findings have highlighted the integral role of oxygen tension in gastrointestinal (GI) homeostasis. During hypoxia when oxygen supply is limited, mammalian cells utilize the Hypoxia-Inducible Factor (HIF) complex to activate genes that promote cell survival and alter glucose metabolism. The GI system exists in a state of physiologic hypoxia, subject to further alterations by pathogens. Although tissue and blood parasites are known to influence tissue oxygen tension, little is known regarding the modulation of hypoxia and HIF by enteric parasites. Aims This project aims to characterize the effect of Giardia duodenalis, a common enteric protozoan that perturbs gastrointestinal function leading to diarrheal disease, on the cellular hypoxic response. We hypothesize that hypoxia-associated genes are activated upon Giardia infection, leading to an adaptive metabolic response with increased glycolytic flux to maintain cellular bioenergetic homeostasis. Methods Caco-2 colonic epithelial cells were infected with Giardia isolate GS/M (MOI 10) for 1.5 or 4.5 hours to capture early and peak HIF activation, under normoxic (21% O2) or hypoxic (1%O2, STEMCELL hypoxia incubator) conditions. RNA was extracted for RT-qPCR to assess transcriptional changes of known HIF-target genes. Intracellular metabolite analysis via liquid-chromatography mass-spectrometry (hydrophilic-interaction chromatography method) was performed to assess HIF-mediated metabolic changes in uninfected and Giardia-infected cells (MOI 10) exposed to the hypoxia mimetic DMOG or the HIF inhibitor PX-478. Results Under normoxic conditions, genes associated with compensatory cellular stress responses such as VEGFA, ANKRD37, GADD45A, and glycolysis-associated genes HK2, and LDHA are upregulated in Giardia-infected cells in a time-dependent manner (pampersand:003C0.05). Under hypoxic conditions, fewer HIF-target genes are upregulated (e.g., VEGFA, GADD45A, PGK1; pampersand:003C0.05), suggesting Giardia-infected cells behave similarly to uninfected cells with constant HIF activation. Analysis of the Caco-2 intracellular metabolome indicates HIF-dependent changes in amino acid (e.g., glycine, threonine) and nucleic acid (e.g., guanosine, inosine, uracil) metabolism. DMOG treatment altered the production of glycolytic intermediates (e.g., DHAP, PEP), confirming the promotion of glycolytic flux by HIF. Conclusions Both cell-stress-related and glucose-metabolism HIF target genes are activated upon Giardia infection. As well, some metabolic changes are dependent on HIF activation. These novel findings indicate Giardia promotes a hypoxic state in human intestinal cells, highlighting a metabolic cell rescue mechanism in response to enteropathogens. Furthermore, understanding the role of hypoxia in enteric parasitic infections will shed light on the potential of HIF as a therapeutic target. Funding Agencies Natural Sciences and Engineering Research Council of Canada (NSERC)

DDX27 regulates oral squamous cell carcinoma development through targeting CSE1L

The headings aimsDEAD-box helicase 27 (DDX27), a member of the DEAD-Box nucleic acid helicase family, holds an elusive role in oral squamous cell carcinoma (OSCC). This study aims to unravel the regulatory functions of DDX27 in OSCC and explore its downstream targets. Materials and methodsA commercial oral squamous cell carcinoma (OSCC) tissue microarray (TMA) was utilized. We analyzed differentially expressed genes in OSCC through the GEO database. Target gene silencing was achieved using the shRNA-mediated lentivirus method. Coexpedia analysis identified co-expressed genes associated with DDX27. Additionally, a Co-Immunoprecipitation (Co-IP) experiment confirmed the protein interaction between DDX27 and CSE1L. Xenograft tumor models were employed to evaluate DDX27's role in OSCC tumor formation. Key findingsElevated DDX27 expression in OSCC correlated with a higher pathological grade. DDX27 knockdown resulted in decreased cell proliferation, increased apoptosis, inhibited cell migration, and induced G2/M phase cell cycle arrest, as well as impaired tumor outgrowth. Coexpedia analysis identified STAU1, NELFCD, and CSE1L as top co-expressed genes. Lentiviral vectors targeting STAU1, NELFCD, and CSE1L revealed that silencing CSE1L significantly impaired cell growth, indicating it as a downstream target of DDX27. Cell rescue experiments demonstrated that increased DDX27 levels ameliorated cell proliferation, attenuated apoptosis, and CSE1L depletion blocked cell development induced by DDX27 overexpression. SignificancesThis study highlighted DDX27 as a potential therapeutic target for OSCC treatment, shedding light on its crucial role in OSCC development. Targeting DDX27 or its downstream effector, CSE1L, holds promise for innovative OSCC therapies.

Mitochondria in Mesenchymal Stem Cells: Key to Fate Determination and Therapeutic Potential.

Mesenchymal stem cells (MSCs) have become popular tool cells in the field of transformation and regenerative medicine due to their function of cell rescue and cell replacement. The dynamically changing mitochondria serve as an energy metabolism factory and signal transduction platform, adapting to different cell states and maintaining normal cell activities. Therefore, a clear understanding of the regulatory mechanism of mitochondria in MSCs is profit for more efficient clinical transformation of stem cells. This review highlights the cutting-edge knowledge regarding mitochondrial biology from the following aspects: mitochondrial morphological dynamics, energy metabolism and signal transduction. The manuscript mainly focuses on mitochondrial mechanistic insights in the whole life course of MSCs, as well as the potential roles played by mitochondria in MSCs treatment of transplantation, for seeking pivotal targets of stem cell fate regulation and stem cell therapy.

Cytoprotective effects of α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, oleic acid and α-tocopherol on 7-ketocholesterol – Induced oxiapoptophagy: Major roles of PI3-K / PDK-1 / Akt signaling pathway and glutathione peroxidase activity in cell rescue

On murine N2a cells, 7-ketocholesterol induced an oxiapotophagic mode of cell death characterized by oxidative stress (reactive oxygen species overproduction on whole cells and at the mitochondrial level; lipid peroxidation), apoptosis induction (caspase-9, −3 and −7 cleavage, PARP degradation) and autophagy (increased ratio LC3-II / LC3-I). Oxidative stress was strongly attenuated by diphenyleneiodonium chloride which inhibits NAD(P)H oxidase. Mitochondrial and peroxisomal morphological and functional changes were also observed. Down regulation of PDK1 / Akt signaling pathways as well as of GSK3 / Mcl-1 and Nrf2 pathways were simultaneously observed in 7-ketocholesterol-induced oxiapoptophagy. These events were prevented by α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, oleic acid and α-tocopherol. The inhibition of the cytoprotection by LY-294002, a PI3-K inhibitor, demonstrated an essential role of PI3-K in cell rescue. The rupture of oxidative stress in 7-ketocholesterol-induced oxiapoptophagy was also associated with important modifications of glutathione peroxidase, superoxide dismutase and catalase activities as well as of glutathione peroxidase-1, superoxide dismutase-1 and catalase level and expression. These events were also counteracted by α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, oleic acid and α-tocopherol. The inhibition of the cytoprotection by mercaptosuccinic acid, a glutathione peroxidase inhibitor, showed an essential role of this enzyme in cell rescue. Altogether, our data support that the reactivation of PI3-K and glutathione peroxidase activities by α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, oleic acid and α-tocopherol are essential to prevent 7KC-induced oxiapoptophagy.

The Management of Children with Pre-School Medulloblastoma – A Review of Four Decades of Multi-Disciplinary and Multi-Institutional Collaboration.

Background: The management and consequent survival and quality of survival for young children with medulloblastoma have undergone substantial improvement over the last four decades. While many of these changes reflect improvements in pediatric neurosurgical, neuro- imaging and radiation oncologic technology and training, as well as in supportive care experience and expertise, this review will focus particularly upon improvements in chemotherapeutic approaches. Methods: This review focuses only upon prospective clinical trials conducted in young children with newly-diagnosed medulloblastoma since the 1980s. The upper age limits for these trials varied from under three to up to six years of age at diagnosis. Results: Certain but not all trials endeavoring to improve outcomes for young children with now- recognized pathological and molecular low-risk characteristics, specifically the Sonic Hedgehog (SHH) sub-type, representing some 50-60% of young children with medulloblastoma, have reaped substantial gains in event-free and overall survival, irradiation-free survival as well as neuropsychological outcomes. Those successful trials utilized either induction including intravenous high-dose methotrexate and intraventricular (intra-Ommaya) methotrexate without consolidation by marrow-ablative chemotherapy with autologous hematopoietic progenitor cell rescue (AuHPCR), or induction with or without intravenous high-dose methotrexate (but not intra-ventricular methotrexate) followed by consolidation with marrow-ablative chemotherapy with AuHPCR. These two approaches have consistently produced superior outcomes compared with other trials. For children with molecularly-characterized Groups 3 or 4 medulloblastoma, representing those at high-risk of relapse, some modest improvements have been made over the years, but much room for improvement remains, especially for those presenting with the highest- risk molecular characteristics (CMYC-amplified Group 3 medulloblastoma) as well as for children with metastatic disease at initial diagnosis. Conclusions: Substantial gains in improving outcomes for young children with medulloblastoma have been achieved with acceptable short- and long-term morbidities of treatment. However, the inclusion of intra-ventricular therapies (conventional chemotherapeutic, radio-labeled monoclonal antibody or adaptive cellular immunotherapeutic approaches) as well as the identification of targeted systemic therapies (biological and/or immunological) and their incorporation into prospective multi-center clinical trials, must be investigated prospectively if we are to improve outcomes for those children at high-risk for relapse, even with the inclusion of refined irradiation therapeutic approaches.

ASB1 inhibits prostate cancer progression by destabilizing CHCHD3 via K48-linked ubiquitination.

Prostate cancer is a major contributor to male mortality worldwide. In this study, we revealed that Ankyrin Repeat and SOCS Box Containing 1 (ASB1) expression was significantly decreased in prostate cancer tissues, correlating strongly with poor patient prognosis. Notably, the group with low ASB1 expression exhibited an increased proportion of M2 macrophages and showed resistance to immune checkpoint inhibitors and cisplatin, but remained sensitive to androgen-receptor-targeting drug bicalutamide. Silencing ASB1 enhanced prostate cancer cell proliferation, clonogenicity, and migration, whereas its overexpression exerted the opposite effects. Through quantitative mass spectrometry interactome analysis, we identified 37 novel proteins interacting with ASB1, including CHCHD3. Subsequent experiments including co-immunoprecipitation, cycloheximide treatment, and ubiquitination assays, revealed that ASB1 interacts with CHCHD3, promoting its degradation via K48-linked ubiquitination. Cell rescue experiments further demonstrated that ASB1 inhibits prostate cancer cell through the CHCHD3/reactive oxygen species (ROS) pathway. Taken together, our study indicated that ASB1 functions as a tumor suppressor by inhibiting CHCHD3/ROS signaling, thereby playing a vital part in prevention of prostate cancer proliferation, clonogenicity, and migration.

An updated cost-effectiveness analysis of axicabtagene ciloleucel in second-line large B-cell lymphoma patients in the United States

Aims This economic evaluation of axicabtagene ciloleucel (axi-cel) versus previous standard of care (SOC; salvage chemotherapy followed by high-dose therapy with autologous stem cell rescue) in the second line (2L) large B-cell lymphoma population is an update of previous economic models that contained immature survival data. Methods This analysis is based on primary overall survival (OS) ZUMA-7 clinical trial data (median follow-up of 47.2 months), from a United States (US) payer perspective, with a model time horizon of 50 years. Mixture cure models were used to extrapolate updated survival data; subsequent treatment data and costs were updated. Patients who remained in the event-free survival state by 5 years were assumed to have achieved long-term remission and not require subsequent treatment. Results Substantial survival and quality of life benefits were observed despite 57% of patients in the SOC arm receiving subsequent cellular therapy: median model-projected (ZUMA-7 trial Kaplan–Meier estimated) OS was 78 months (median not reached) for axi-cel versus 25 months (31 months) for SOC, resulting in incremental quality-adjusted life year (QALY) difference of 1.63 in favor of axi-cel. Incrementally higher subsequent treatment costs were observed in the SOC arm due to substantial crossover to cellular therapies, thus, when considering the generally accepted willingness to pay threshold of $150,000 per QALY in the US, axi-cel was cost-effective with an incremental cost-effectiveness ratio of $98,040 per QALY. Conclusions Results remained consistent across a wide range of sensitivity and scenario analysis, including a crossover adjusted analysis, suggesting that the mature OS data has significantly reduced the uncertainty of axi-cel’s cost-effectiveness in the 2L setting in the US. Deferring treatment with CAR T therapies after attempting a path to transplant may result in excess mortality, lower quality of life and would be an inefficient use of resources relative to 2L axi-cel.

Long-term Outcomes of Protocol-Based Treatment for Newly Diagnosed Medulloblastoma.

The Korean Society of Pediatric Neuro-Oncology (KSPNO) conducted treatment strategies for children with medulloblastoma (MB) by using alkylating agents for maintenance chemotherapy or tandem high-dose chemotherapy (HDC) with autologous stem cell rescue (ASCR) according to the risk stratification. The purpose of the study was to assess treatment outcomes and complications based on risk-adapted treatment and HDC. Fifty-nine patients diagnosed with MB were enrolled in this study. Patients in the standard-risk (SR) group received radiotherapy (RT) after surgery and chemotherapy using the KSPNO M051 regimen. Patients in the high-risk (HR) group received two and four chemotherapy cycles according to the KSPNO S081 protocol before and after reduced RT for age following surgery and two cycles of tandem HDC with ASCR consolidation treatment. In the SR group, 24 patients showed 5-year event-free survival (EFS) and overall survival (OS) estimates of 86.7% (95% confidence interval [CI], 73.6 to 100) and 95.8% (95% CI, 88.2 to 100), respectively. In the HR group, more infectious complications and mortality occurred during the second HDC than during the first. In the HR group, the 5-year EFS and OS estimates were 65.5% (95% CI, 51.4 to 83.4) and 72.3% (95% CI, 58.4 to 89.6), respectively. High intensity of alkylating agents for SR resulted in similar outcomes but with a high incidence of hematologic toxicity. Tandem HDC with ASCR for HR induced favorable EFS and OS estimates compared to those reported previously. However, infectious complications and treatment-related mortalities suggest that a reduced chemotherapy dose is necessary, especially for the second HDC.

Treated Non Seminomatous Germ Cell Tumor of Testes Post Resection of Teratomatous Lung Nodule

Background: Testicular tumors are relatively rare, accounting for only 1% of all malignancies in men. Among the different types of testicular tumors, germ cell tumors are the most common. They are very sensitive to chemotherapy and are curable even when metastatic. Cure rates approximates 90%-95% in good risk and early stage tumors. Earlier on, metastatic testicular cancer was usually incurable and fatal, but newer treatments including high-dose chemotherapy and stem cell rescue have changed the scenario. Case Presentation: This case report presents a case of a 30-year-old male with a large testicular tumor, diagnosed as a mixed germ cell tumor consisting of a yolk sac tumor, teratoma with immature elements, and seminoma with germ cell neoplasia in situ. After undergoing radical orchiectomy, the patient experienced a postoperative decline in alpha-fetoprotein levels and was placed on surveillance. Subsequent imaging revealed the development of a lung nodule, accompanied by an increase in alpha-fetoprotein levels. The patient received chemotherapy with three cycles of BEP (bleomycin, etoposide, platinum), resulting in a partial response. Due to the patient's COVID-19 infection, the final cycle of chemotherapy was modified. Further imaging showed a marginal decrease in the size of the lung nodule, prompting surgical resection. The lung nodule was found to be a post-pubertal teratoma without significant immature elements. Following surgery, the patient's alpha-fetoprotein levels decreased to a baseline nadir. Close follow-up continues, demonstrating no evidence of disease recurrence. Conclusion: This case emphasizes the importance of a multidisciplinary approach, including surgery and chemotherapy, in managing testicular germ cell tumors with metastasis, leading to favorable outcomes and long-term disease control.

Bone Marrow Failure Associated with Short Telomeres and Digenic Variants of Uncertain Significance in Telomere Biology Genes

DISCUSSION Telomere Biology Disorders (TBD) are a group of heritable disorders characterized by abnormally short telomeres. There are at least 15 genes associated with TBD including ACD, CTC1, DKC1, NHP2, NOP10, PARN, POT1, RTEL1, STN1, TERC, TERT, TINF2, WRAP53 NAF1 and ZCCHC8, which can be inherited in autosomal dominant (AD), autosomal recessive (AR), or X-linked recessive patterns. Genetic anticipation has been described in some families with more severe symptoms appearing in each successive generation. We report two patients that presented with bone marrow failure, who were subsequently identified to have short telomeres and variants of uncertain significance (VUS) in two different telomere genes inherited from their parents. These cases suggest a potential for digenic inheritance of TBD. CASE 1: At age 6, patient was diagnosed with stage 4 neuroblastoma. He was treated with chemotherapy, surgery, immunotherapy, and autologous stem cell rescue. At age 10, he developed frequent and severe epistaxis and was found to have pancytopenia. He underwent a bone marrow aspirate and biopsy which revealed a hypocellular marrow and der (1;7), which is associated with myelodysplastic syndrome (MDS). Genetic evaluation revealed one pathogenic variant in DNAJC21 (from father) and VUS in NAF1 (c.1375C>T) (from father) and RTEL1 (c.533T>C) (from mother). Because NAF1 and RTEL1 are related to AD or AR TBD, telomere length testing was performed. Patient was found to have low telomere length, < 1st percentile, in granulocytes, lymphocytes, memory T cells and NK cells (Figure 1). The patient's mother was found to have borderline low telomere length >=1 and <10 th percentiles in her lymphocytes, naïve T cells, memory T cells, and B cells, and low telomere length in her granulocytes. Father had borderline low telomere length >=1 and <10 th percentiles in granulocytes but normal telomere length in all other subsets. Neither parent reported any symptoms that would be consistent with TBD. We interpreted these findings as consistent with the patient having TBD and the patient's mother either also having TBD or being a carrier, and the potential that genetic anticipation may be involved. The NAF1 variant appears likely to be benign as father's telomeres were normal in length except for low telomere length in his granulocyte compartment. It is possible that the patient's prior treatment for neuroblastoma may have contributed to the shortened telomeres. Over the course of a year, patient developed worsening bone marrow failure. He then underwent reduced intensity conditioning using busulfan, fludarabine and anti-thymocyte globulin for a 4/6 matched cord stem cell transplant. He engrafted on day +10. CASE 2: A 15-year-old male presented with a depressed skull fracture and scalp hematoma following a sports-related head injury. He was incidentally found to have a platelet count of 69 x 10 3/microliter, mild anemia with macrocytosis, and mild leukopenia. Bone marrow biopsy revealed a hypocellular marrow with multilineage maturation, mild fibrosis, and no cytogenetic abnormalities. The patient's telomere lengths were low, below the 1 st percentile, in all 6 white blood cell subsets (Figure 2). He underwent clinical trio exome sequencing along with his parents. He was found to have VUSs in TERT (c.3158-80G>A) (from mother), TINF2 (c.814T>C) (from mother) and SRP72 (c.1928C>T) (from father). Mother was also found to have low telomere length (<1 st percentile) in all 6 subsets. Both parents have normal blood counts while patient continues to have pancytopenia and is transfusion independent. It is presently unclear which of the VUSs are contributing to the patient's phenotype as the TERT and TINF2 variants are both associated with bone marrow failure and heterozygous germline variants in SRP72 is associated with familial MDS. Genes that cause TBD can be inherited through various modes of inheritance. These two patients who presented with bone marrow failure, identified to have VUSs in more than one TBD-associated genes and functional evidence of shortened telomeres, highlight the potential for a digenic mode of inheritance underlying TBD. Synergy between two variants contributes to a penetrant phenotype and can result in earlier or more severe onset of disease. Our patients' parents were asymptomatic or mildly affected, compared to our patients. Larger cohort studies are needed to determine the prevalence of this mode of inheritance.

RADAR Trial: MRD Response Adapted Trial for Newly Diagnosed Transplant Eligible Myeloma Patients

Background: Achieving and sustaining minimal disease negativity improves clinical outcomes in newly diagnosed transplant eligible myeloma (NDMM). Outcomes in NDMM are influenced by levels of minimal residual disease (MRD) post therapy and genetic risk by FISH or molecular profiling prior to therapy. Trials exploring response adapted post transplant strategies using MRD as a prognostic tool are going. We provide an update of the current UK-MRA RADAR study. Study Design/Methods: The UK-MRA RADAR study is a national multicentre risk adapted response guided multiarm multistage (MAMS) phase II/III trial which will recruit 1400 patients with NDMM eligible for ASCT. Eligible patients receive induction with 4 cycles of RCyBorD (lenalidomide, cyclophosphamide, bortezomib, dexamethasone), followed by high-dose melphalan and stem cell rescue. Patients with high-risk disease receive Isatuximab in addition to RCyBorD during induction. Genetic risk classification is performed using standard of care FISH testing at local sites. Participants enter the standard or high risk pathway based on presence of atleast two markers: t(4;14), t(14;16), t(14;20), del(17p), del(1p) and gain (1q21). MRD analysis in RADAR uses a multiparametric flow cytometry panel with CD319 and Cd229 used to augment post-isatuximab sample analysis at a central laboratory, with negativity defined at threshold of 10 -5(Figure 1). For standard risk patients, the study addresses two pivotal MRD-related questions: 1) For those with sustained MRD negativity after 12 months of isatuximab maintenance, can treatment be stopped safely to allow patients a treatment-free interval thereby reducing treatment burden? 2) For patients with persistent detectable disease after ASCT, does intensification of consolidation and maintenance therapy augment conversion to MRD negativity compared to lenalidomide maintenance? High risk participants receive intensified consolidation and maintenance regimens post_ASCT aligned to OPTIMUM/MUKnine ( NCT03188172). A further amendment is planned to explore the activity of a BCMA bispecific antibody in high risk patients pre and post transplant. It is hypothesised this will improve both depth (MRD negativity) and ability to sustain response (PFS). Feasibility and benefits of multimodal MRD incorporating imaging and blood-based testing will be tested in the new high risk pathway. In RADAR, we will also gather information on how patients respond to receiving information about their genetic risk assignment and treatment response (MRD). This is a unique opportunity to understand the patients perspective on personalised medicine, which has implications for implementation in clinical care, and future trial design. Trial Progress. RADAR opened to recruitment in May 2021 and is currently recruiting across 87 hospital sites with a further 16 sites in set up. 504 patients have been recruited; 87 are classified as high risk on FISH analysis ( 375 standard risk, 15 unable to determine, 19TBC, 8 withdrawn before FISH analysis). 220 have reached D+100 MRD assessment. Trackable phenotypes were available in 98.2% of patients. Discussion: The RADAR study simultaneously tests escalation and de-escalation strategies, according to disease response, and using MRD to guide post transplant therapy in NDMM patients. Use of central flow cytometry laboratory for MRD has proven feasible in patients to date. Recruitment is on target and outcomes will provide RCT-level evidence for the adaptive use of MRD in treatment allocation and intensification post transplant. The platform design enables adaptation with ability to test new strategies for high risk patients and for standard risk patients who are MRD positive post-transplant. Funding This trial is funded by Cancer Research UK (C9203/A24078), Sanofi Genzyme and Celgene : A BMS company. Sanofi Genzyme and Celgene: A BMS company provide study drug and unrestricted educational grants to support study coordination. This work is also supported by Core Clinical trials unit infrastructure from Cancer Research UK ( C7852/A25447). The funder of the study had no role in study design, data collection, data analysis, data interpretation or writing of the abstract