RaNA Therapeutics (United States)

Recent advances in fibroblast activation protein imaging have led to the development of new fibroblast activation protein-targeting radiotracers. This study investigates the mass dose effects of [68Ga]Ga-LNTH-1363S on image quality. Methods: In this retrospective evaluation of 39 patients with various oncologic diseases (including breast cancer and sarcoma), different administered mass doses (30, 75, 90, and 100 μg) of [68Ga]Ga-LNTH-1363S were visually and quantitatively (blood pool SUVmean, lesion SUVmax, and tumor-to-blood pool ratios [TBRs]) assessed. Results: Lower peptide masses were associated with prominent blood pooling in [68Ga]Ga-LNTH-1363S PET imaging and reduced TBRs, whereas higher mass doses yielded superior image quality because of lower mean tracer accumulation in the blood pool and therefore higher TBRs. Conclusion: These observations provide evidence that biodistribution and imaging quality are dependent on the administered mass dose. Further studies are warranted to validate these findings.

KRAS G12D is a common oncogenic driver mutation in diverse cancers, including non-small cell lung cancer, colorectal cancer, and pancreatic cancer. KRASG12D inhibitors have recently progressed into clinical trials but will likely face innate or acquired drug resistance similar to that which has been observed for KRASG12C inhibitors, such as activation of receptor tyrosine kinases, KRAS independence, and reactivation of RAS-MAPK signaling. This study investigates heterobifunctional small-molecule dual inhibitors that simultaneously target both KRASG12D and protein chaperone HSP90 in KRASG12D-mutated cancer cell lines and patient-derived organoids. Our findings reveal that the efficacy of the clinical-stage KRASG12D inhibitor MRTX1133 varies, with notable resistance being observed in some cell line and organoid models. In contrast, KRASG12D-HSP90 dual inhibitors were found to broadly display superior effectiveness in inducing apoptosis, reducing cell viability, and suppressing key downstream signaling pathways such as AKT and ERK1/2 in MRTX1133-resistant models. The rationale for targeting HSP90, which is preferentially activated in cancer cells, alongside KRASG12D, arises from the ability of HSP90 inhibition to destabilize substrate client proteins that are essential for cancer cell survival and have also been implicated in resistance to KRAS inhibitors. This dual-inhibitor approach presents a promising new strategy to combat de novo and acquired drug resistance in KRASG12D-mutated cancers and potentially paves the way for improved clinical outcomes by addressing the complex molecular mechanisms underlying cancer cell evolution that enables resistance to conventional inhibitors.

Purpose:The development of DNA damage response (DDR)-directed therapies is a major area of clinical investigation; however, to date, PARP inhibitors (PARPi) remain the only approved therapy in this space. Major challenges to DDR-targeted therapies in the post-PARPi therapy era are the context dependency of DDR alterations and the presence of preexisting resistance in this heavily pretreated population. Blood samples from patients with tumors harboring defects in DDR genes were used to evaluate the feasibility of a liquid biopsy platform to detect complex genomic events such as BRCA1/2 reversions, homologous recombination deficiency (HRD) signatures, pathogenic variant allele status, and differentially methylated regions for accurate quantitation of tumor fraction.Experimental Design:Pretreatment ctDNA samples from 173 patients enrolled in two phase 1/2 clinical trials (TRESR; NCT04497116 and ATTACC; NCT04972110) were selected for analysis. Results:In a phase I heavily pretreated patient population with DDR defects, complex genomic alterations (HRD, biallelic loss, and complex reversions) that historically require tumor tissue biopsies could be detected in ctDNA. Within the cohort of BRCA-associated tumor types previously treated with PARPi or platinum therapy, HRD reversions were detected in 44% of evaluable patients and included large genomic rearrangements leading to deletion of whole or partial exons which have been underrepresented in the literature because of technological limitations.Conclusions:This study showcases the genomic complexity of DDR-altered tumors as revealed through baseline ctDNA profiling, an understanding of which is crucial for the future clinical development of novel DDR-directed therapies and combinations.

Abstract We previously discovered that BM-derived Mesenchymal stromal cells (BM-MSCs) are migrating to the stroma of numerous cancers and their metastasis, forming TAFs and secrete proteins intra-tumorally. MSCs have not been used in cancer therapy because of their immune suppressive properties, limited replicative capacity, source dependent quality and controversial roles in cancer biology. Here, we report the characterization of novel MSCs (iMSCs), which were generated from adult skin fibroblast-derived induced pluripotent stem cells (iPSC). These iPSCs were generated via a unique, transient mRNA transfection technique and subsequently differentiated into iMSCs. Notably, iMSCs displayed contact inhibition and significantly enhanced proliferative capacity under both normoxic and hypoxic conditions, outperforming bone marrow-derived MSCs (BM-MSCs) while fully retaining trilineage differentiation potential. Comprehensive molecular profiling, including RNA sequencing, single-cell mass cytometry (CyTOF), and Luminex assays, revealed strong phenotypic and functional resemblance between iMSCs and BM-MSCs. Crucially, no sarcoma formation was detected in NSGS mice following intraperitoneal, subcutaneous, or intravenous administration of iMSCs, underscoring their safety profile. We further engineered a DNA cassette into these cells to enable IL-7 and IL-15 cytokine production, expressed as either individual molecules (P2A) or a single fused molecule (FUS). Both P2A and FUS cells demonstrated the capacity to drive T cell proliferation autonomously in co-culture. Cytokine secretion from these modified iMSCs enhanced tumor cell death in a triple co-culture system comprising iMSCs, the ovarian cancer cell line ID8, and human PBMCs. In a syngeneic mouse model of ovarian cancer (ID8 in C57BL/6 mice), administration of P2A or FUS MSCs resulted in reduced tumor growth and extended survival. Immunohistochemical and flow cytometric analyses revealed massive infiltration of T cells, macrophages, NK cells, and other immune cells into the tumor microenvironment (TME) in both FUS or P2A groups, but not in control, unmodified MSC, or PBS injected animals. Moreover, the TME in P2A- and FUS-treated mice showed enrichment in M1-type macrophages, with no detection of any of the T cell exhaustion markers tested, in contrast to the control groups (PBS or GFP-iMSC). In conclusion, our findings indicate that IL7-IL15-secreting iMSCs migrate into solid tumors, induce massive immune cell infiltration into the TME and enhance antitumor immunity in a syngeneic mouse model of cancer. These cytokine-producing iMSCs represent a promising therapeutic strategy for modulating the immune response against tumors and advancing anticancer immunotherapy by converting “cold” into “hot” tumor microenvironments. Citation Format: Sandeep Singh, Andrea Bedoy, Dipmoy Nath, Kyung Hee Chang, Lauren B. Ostermann, Ivo Veletic, Christopher D. Pacheco, Po Yee Mak, Edward Ayoub, Muharrem Muftuoglu, Mahesh Basyal, Taeyun Kim, Raven Dance Hinkel, Kyle M. Garland, Matthew Angel, Sanjeev Luther, Christopher B. Rohde, Michael Andreeff. iPS-derived MSC secreting IL7 and IL15 exert immune mediated anti-tumor activities [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3473.

BackgroundMitochondrial dysfunction is widely reported in models of Huntington’s disease (HD), including altered respiratory complex activity, altered mitochondrial membrane potential and changes in mitophagy. However, whether mitochondrial function could be rescued as a therapeutic target in HD is under explored. This study sought to determine whether HD donor-derived fibroblasts could be used to identify modulators of mitochondrial function with therapeutic potential.MethodsFibroblasts from 3 HD donors and 3 age-matched control donors were characterised for respiration, and total cellular ATP levels. High content imaging was used to assess multiple parameters of mitochondrial function, mitochondrial morphology and mitophagy flux. Tool compounds predicted to affect mitochondrial function were selected for screening using high throughput imaging.ResultsMitochondria from HD donor-derived fibroblasts display similar ATP levels and basal respiration to controls but significantly lower maximal respiration under uncoupling conditions. High content imaging identified alterations in mitochondrial morphology that were consistent across all HD cell lines and alterations in number and mitochondrial membrane potential that are specific to individual patient cell lines. HD donor-derived fibroblasts displayed altered mitophagy flux under mitophagy inducing conditions. Proof-of-principle live imaging screening identified A769662 which can enhance the proportion of functional mitochondria in HD donor-derived fibroblasts.ConclusionsThis study has identified alterations in HD donor-derived fibroblasts in mitochondrial morphology, function, and clearance, indicating multiple potential target areas for screening for therapeutic intervention. Proof-of-principle screening identified a modulator of mitochondrial function, confirming the potential of using HD donor-derived fibroblasts with high content imaging to identify mitochondrial therapeutics for HD.

This study aimed to purify and characterize immunoregulatory peptides from Sipunculus nudus L. and to explore the underlying mechanisms. Ultrafiltration, gel filtration chromatography, and reverse phase high-performance liquid chromatography (RP-HPLC) were used to purify the peptide following enzymatic hydrolysis. Rates of lymphocyte proliferation and phagocytosis as well as nitric oxide (NO) production levels were used as indicators of immunoregulatory activity to screen the fractions. The amino acid sequence of the peptide, designated as SNLP, was identified as Arg-Val-Lys-Gly-Lys-Ile-Leu-Ala-Lys-Arg-Leu-Asn (RVKGKILAKRLN) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Treatment with the synthetic SNLP increased the proliferation and phagocytosis of RAW 264.7 macrophages and promoted the secretion of tumor necrosis factor-ɑ (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β), and NO levels. The mRNA levels of these cytokines and iNOS were also increased by SNLP. Our results provide preliminary evidence suggesting that SNLP acts as a dual immunomodulatory peptide with immunostimulatory and anti-inflammatory activities. In summary, SNLP derived from Sipunculus nudus L. is a potent immunoregulatory peptide and represents a potential functional food or immunoregulatory drug.

Intraovarian growth factors play a vital role in influencing the fate of ovarian follicles. They affect proliferation and apoptosis of granulosa cells (GC) and can influence whether small antral follicles continue their growth or undergo atresia. Transforming growth factor-alpha (TGFα), an oocyte-derived growth factor, is thought to regulate granulosa cell function; yet its investigation has been largely overshadowed by emerging interest in TGF-beta superfamily members, such as bone morphogenetic proteins (BMP) and anti-Mullerian hormone (AMH). Here, effects of TGFα on bovine GC proliferation, intracellular signaling, and cytokine-induced apoptosis were evaluated. Briefly, all small antral follicles (3-5 mm) from slaughterhouse specimens of bovine ovary pairs were aspirated and the cells were plated in T25 flasks containing DMEM/F12 medium, 10% FBS, and antibiotic-antimycotic, and incubated at 37 °C in 5% CO2 for 3 to 4 d. Once confluent, the cells were sub-cultured for experiments (in 96-, 12-, or 6-well plates) in serum-free conditions (DMEM/F12 medium with ITS). Exposure of the bGC to TGFα (10 or 100 ng/mL) for 24 h stimulated cell proliferation compared to control (P < 0.05; n = 7 ovary pairs). Proliferation was accompanied by a concomitant increase in mitogen-activated protein kinase (MAPK) signaling within 2 h of treatment, as evidenced by phosphorylated ERK1/2 expression (P < 0.05, n = 3 ovary pairs). These effects were entirely negated, however, by the MAPK inhibitor, U0126 (10uM, P < 0.05). Additionally, prior exposure of the bGC to TGFα (100 ng/mL) failed to prevent Fas Ligand (100 ng/mL)-induced apoptosis, as measured by caspase 3/7 activity (P < 0.05, n = 7 ovary pairs). Collectively, the results indicate TGFα stimulates proliferation of bGC from small antral follicles via a MAPK/ERK-mediated mechanism, but this action alone fails to prevent apoptosis, suggesting that TGFα may be incapable of promoting their persistence in follicles during the process of follicular selection/dominance.