Akebia Therapeutics (United States)

<p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d627885e81"> <b>Background:</b> In vivo engineering of CAR T cells using lipid nanoparticles (LNPs) [ <a class="xref-link" href="#r1">1</a>] represents a transformative therapeutic modality, offering scalable, non-viral delivery of mRNA to immune cells. We have developed a CD8+ T-cell-targeted LNP (tLNP) encapsulating mRNA encoding a CD20-directed chimeric antigen receptor (CAR) as a surrogate system to demonstrate pharmacologic effect in non-human primates (NHPs). In NHPs, systemic tLNP administration resulted in transient CAR expression on endogenous CD8+ T cells and subsequent depletion of CD20+ B cells in both peripheral blood and spleen. To characterize the in vivo dynamics and inform translational dosing strategies, we developed a pharmacokinetic/cellular kinetic/pharmacodynamic (PK/CK/PD) model [ <a class="xref-link" href="#r2">2</a>, <a class="xref-link" href="#r3">3</a>]. <p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d627885e95"> <b>Methods:</b> NHPs were administered multiple doses of tLNPs. The modeling framework integrated: Plasma mRNA pharmacokinetics (PK), CAR expression kinetics (CK) in circulating T cells, and Pharmacodynamic (PD) impact on CD20+ B-cell depletion. CAR expression was modeled as a function of mRNA exposure, with downstream PD effects driven by CAR T-cell-mediated B-cell cytotoxicity in blood and spleen. CAR T-cell clearance incorporated both first-order elimination and a target-mediated CAR-T clearance (TMCC) pathway reflective of CAR/B-cell interaction. <p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d627885e100"> <b>Results:</b> The model robustly recapitulated observed in vivo data. mRNA PK profiles, consistent across single and repeat dosing, were well captured. CAR expression was transient and scaled linearly with mRNA exposure. Importantly, CAR T-cell clearance dynamics were dose- and exposure-dependent. Initial doses exhibited rapid clearance via TMCC due to high B-cell burden. Subsequent doses, administered post-partial B-cell depletion, exhibited reduced TMCC and prolonged CAR T-cell persistence. This dynamic translated into more durable and extensive B-cell depletion upon repeat dosing. <p xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" dir="auto" id="d627885e105"> <b>Conclusions:</b> Developed semi-mechanistic model elucidates the exposure-response relationship of in vivo engineered CAR T cells following LNP-mediated mRNA delivery. The framework enables simulation of dosing regimens and supports optimization of LNP dose levels for maximal B-cell depletion. These insights guide preclinical development and lay the groundwork for rational clinical translation of the mRNA-LNP in vivo CAR-T platform—a novel, modular approach to programmable immunotherapy.

Abstract Background and Aims Anaemia of chronic kidney disease (CKD) is a major unmet medical need. Vadadustat, a hypoxia-inducible factor prolyl hydroxylase inhibitor (HIF-PHI), approved for the treatment of anaemia of dialysis-dependent CKD, stimulates endogenous erythropoietin (EPO) production, increasing haemoglobin (Hb) and haematocrit (HCT). Ferric citrate is a source of iron and is used as a treatment for iron deficiency anaemia. The objectives of these studies were to determine if the co-administration of vadadustat with a well-absorbed oral iron preparation such as ferric citrate would have additive effects on anaemia correction. Method The experiments were conducted in the peptidoglycan polysaccharide (PGPS) rat model of inflammation-induced anaemia, and the rat model of adenine-induced CKD (n = 10/group). Primary treatments included both vadadustat and ferric citrate alone and in combination, with appropriate controls. Recombinant human EPO (rhEPO) was used in some experiments as a comparator. Results In the PGPS rat model of inflammation-induced anaemia, the coadministration of vadadustat and ferric citrate caused a statistically significant increase in serum iron and transferrin saturation (TSAT) and a statistically significant decrease in total iron binding capacity (TIBC) compared to when vadadustat or ferric citrate were given alone. There were also increases in Hb, HCT, percent reticulocytes, mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) levels. Regarding inflammation, the combination of vadadustat and ferric citrate treatment decreased white blood cell counts in a statistically significantly manner and this was associated with decreases in neutrophil and lymphocyte counts, haptoglobin, IL-6, IL-15, C-reactive protein, ankle diameter and paw weight. Similarly, in the rat model of adenine-induced CKD, there was a statistically significant increase in serum iron levels in the rats given vadadustat in combination with ferric citrate compared to the rats given ferric citrate or vadadustat alone or in rats given rhEPO either alone or in combination with ferric citrate. Consistent with the serum iron data, the combination of vadadustat and ferric citrate resulted in increases in TIBC, serum iron saturation percent, Hb, HCT, MCV and MCH levels. Serum levels of IL-6, FGF-21, and myostatin, markers of inflammation, did not differ statistically across groups. Conclusion Collectively, these data suggest vadadustat in combination with oral iron provides greater improvement in anaemia related parameters in models of inflammation-induced anaemia and CKD than when either agent is given alone.

ABSTRACT The annual ImmunoRad Conference has established itself as a recurrent occasion to explore the possibility of combining radiation therapy (RT) and immunotherapy (IT) for clinical cancer management. Bringing together a number of preclinical and clinical leaders in the fields of radiation oncology, immuno-oncology and IT, this annual event fosters indeed essential conversations and fruitful exchanges on how to address existing challenges to expand the therapeutic value of RT-IT combinations. The 8th edition of the ImmunoRad Conference, which has been held in October 2024 at the Weill Cornell Medical College of New York City, highlighted exciting preclinical and clinical advances at the interface between RT and IT, setting the stage for extra progress toward extended benefits for patients with an increasing variety of tumor types. Here, we critically summarize the lines of investigation that have been discussed at the occasion of the 8th Annual ImmunoRad Conference.

IL-12 mediates innate and adaptive immune responses and has demonstrated therapeutic antitumor activity, but clinical development has been hindered by a narrow therapeutic window. We generated a novel IL-12-anchored drug conjugate by physiochemical linking of murine IL-12 to aluminum hydroxide (alum). The complex was designed to utilize alum as a scaffolding for durable retention of IL-12 within the tumor microenvironment as a strategy to increase the therapeutic window. To better define the systemic pharmacokinetic (PK) profile of the anchored IL-12 (mANK-101), a model-based assessment tool was developed to describe the systemic PK profile and downstream signaling factors following intratumoral injection of mANK-101. When compared with nonanchored IL-12, mANK-101 exhibited a distinct PK profile. Specifically, mANK-101 treatment was associated with a significant ninefold increase in the systemic terminal volume of distribution (Vd). Furthermore, linear mixed-effects models provided evidence that CD8+ T-cell infiltration and increased serum IFN-γ levels were correlated with tumor regression after a single dose of mANK-101. In addition, PK/pharmacodynamic modeling confirmed a link between systemic IL-12 and serum IFN-γ. The model also suggests that the anchored IL-12 drug conjugate is expected to prolong the absorption half-life (115 hours vs. 8 hours for the unanchored drug) with durable local retention and limited systemic absorption. In addition, serum IFN-γ may be a surrogate marker for drug activity. The PK modeling predictions may also contribute to determining the optimal clinical dose and schedule of mANK-101 and other anchored drug conjugates in patients with solid tumors.

Vadadustat is an oral hypoxia-inducible factor prolyl hydroxylase inhibitor for treating anemia in chronic kidney disease (CKD). Single ascending dose (SAD) and multiple ascending dose (MAD) studies assessed pharmacokinetics (PK), pharmacodynamics (PD), and safety of vadadustat in healthy volunteers. A single-dose, open-label study was conducted in patients with CKD stages 3 and 4 not on dialysis. In the SAD study, 48 healthy adult men (n = 8/cohort) received single doses of vadadustat (80-1,200 mg) or placebo. In the MAD study, 34 healthy adult men (n = 8-9/cohort) received daily vadadustat (500-900 mg) or placebo for 10 days. In the single-dose CKD study, 22 male and female patients with CKD (stage 3: n = 10; stage 4: n = 12) received single doses of vadadustat (500 mg). PK parameters included plasma vadadustat; PD biomarkers were measured, including erythropoietin (EPO) levels, reticulocytes, and others. Plasma vadadustat peaked 3-4 hours after single or multiple dosing in healthy volunteers, with a mean t 1/2 of approximately 4.5 hours. In patients with CKD, plasma vadadustat peaked at 5-6 hours, with a mean t 1/2 of 7.2 (stage 3) and 8.5 (stage 4) hours. Vadadustat AUC∞ and C max increased dose proportionally in SAD and MAD trials. In all trials, EPO concentrations increased in a dose-related manner and returned approximately to baseline by 24 hours. Adverse events were mild and considered not study drug related. The PK and PD results of these studies were utilized for further clinical development of vadadustat for treatment of anemia in CKD patients.

Cancer immunotherapy has flourished over the last 10–15 years, transforming the practice of oncology and providing long-term clinical benefit to some patients. During this time, three distinct classes of immune checkpoint inhibitors, chimeric antigen receptor-T cell therapies specific for two targets, and two distinct classes of bispecific T cell engagers, a vaccine, and an oncolytic virus have joined cytokines as a standard of cancer care. At the same time, scientific progress has delivered vast amounts of new knowledge. For example, advances in technologies such as single-cell sequencing and spatial transcriptomics have provided deep insights into the immunobiology of the tumor microenvironment. With this rapid clinical and scientific progress, the field of cancer immunotherapy is currently at a critical inflection point, with potential for exponential growth over the next decade. Recognizing this, the Society for Immunotherapy of Cancer convened a diverse group of experts in cancer immunotherapy representing academia, the pharmaceutical and biotechnology industries, patient advocacy, and the regulatory community to identify current opportunities and challenges with the goal of prioritizing areas with the highest potential for clinical impact. The consensus group identified seven high-priority areas of current opportunity for the field: mechanisms of antitumor activity and toxicity; mechanisms of drug resistance; biomarkers and biospecimens; unique aspects of novel therapeutics; host and environmental interactions; premalignant immunity, immune interception, and immunoprevention; and clinical trial design, endpoints, and conduct. Additionally, potential roadblocks to progress were discussed, and several topics were identified as cross-cutting tools for optimization, each with potential to impact multiple scientific priority areas. These cross-cutting tools include preclinical models, data curation and sharing, biopsies and biospecimens, diversification of funding sources, definitions and standards, and patient engagement. Finally, three key guiding principles were identified that will both optimize and maximize progress in the field. These include engaging the patient community; cultivating diversity, equity, inclusion, and accessibility; and leveraging the power of artificial intelligence to accelerate progress. Here, we present the outcomes of these discussions as a strategic vision to galvanize the field for the next decade of exponential progress in cancer immunotherapy.

Abstract Background: The composition of the tumor microenvironment (TME) is a major determinant of response to therapy in many solid tumors. To-date, characterization of the TME has been based on limited analysis of thin tissue sections. Here we demonstrate the utility of an end-to-end 3D spatial biology workflow, the 3D I/O Pro™, based on whole tissue imaging, to identify and quantify tumor cells, lymphocytes, and collagen features in human FFPE tissue samples. Methods: Five human colorectal cancer FFPE blocks were deparaffinized, stained with nuclear (TO-PRO-3) and general protein (eosin) dyes, and cleared using a modified iDISCO protocol. Entire samples were imaged at 2 µm/pixel resolution with a hybrid open-top light-sheet microscope, the 3Di™. Regions of interest (ROIs) with a volume of 0.5 mm3 were re-imaged at 0.33 µm/pixel. Cell nuclei and collagen were segmented and 3D spatial relationships between tumor cells, lymphocytes, and collagen fibers were quantified. Analyses were performed within select ROIs using 3Dai™ tools including U-Net, CytoMAP, CT-FIRE, CurveAlign, and custom Python scripts. Results: The ratio of stromal to tumor parenchymal lymphocytes (lymphocyte infiltration ratio) varied from 1.4 up to 9.1 in 3D volumes and 1 to 25 in 2D virtual sections taken throughout all 5 CRC samples. The ratio of the collagen fibers within 100 µm of the tumor border-oriented perpendicular to the tumor-stromal boundary compared to those oriented parallel to the tumor-stromal boundary was 0.37 in a proof-of-concept evaluation of a 2D virtual section with a lymphocyte infiltration ratio of 12. Qualitatively, areas with perpendicular collagen had more lymphocyte infiltration into the tumor parenchyma than areas with parallel collagen orientation. Conclusions: We demonstrated that the 3D I/O Pro™ pipeline can quantify lymphocyte density in tumor parenchyma and stroma and analyze collagen features, including orientation, within 3D ROIs in the TME. This workflow allows us to characterize tumors based on many complex spatial relationships and could have broad applicability in research and development of novel cancer therapies that target tumor fibrosis or other features of the TME. In the future, we plan to correlate features from the 3D TME with response to immunotherapy and use these features to refine histologic definitions of immune spatial phenotypes. Citation Format: Caleb Stoltzfus, Jasmine Wilson, Nathan Grant, Alexandra Alvarsson, Anna DeWitt, David Simmons, Brandy Olin, Bonnie Phillips, Laura A. Dillon, G. Travis Clifton, Nicholas P. Reder. 3D spatial quantification of lymphocyte infiltration and collagen features in the tumor microenvironment using a novel assay: 3D I/O Pro™ [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 2309.

Invasive cancers typically evade immune surveillance through profound local and systemic immunosuppression, preventing their elimination or control. Targeting immune interventions to prevent or intercept premalignant lesions, before significant immune dysregulation has occurred, may be a more successful strategy. The field of cancer immune interception and prevention is nascent, and the scientific community has been slow to embrace this potentially most rational approach to reducing the global burden of cancer. This may change due to recent promising advances in cancer immunoprevention including the use of vaccines for the prevention of viral cancers, the use of cancer-associated antigen vaccines in the setting of precancers, and the development of cancer-preventative vaccines for high-risk individuals who are healthy but carry cancer-associated heritable genetic mutations. Furthermore, there is increasing recognition of the importance of cancer prevention and interception by national cancer organizations. The National Cancer Institute (NCI) recently released the National Cancer Plan, which includes cancer prevention among the top priorities of the institute. The NCI’s Division of Cancer Prevention has been introducing new funding opportunities for scientists with an interest in the field of cancer prevention: The Cancer Prevention-Interception Targeted Agent Discovery Program and The Cancer Immunoprevention Network. Moreover, the Human Tumor Atlas Network is spearheading the development of a precancer atlas to better understand the biology of pre-invasive changes, including the tissue microenvironment and the underlying genetics that drive carcinogenesis. These data will inform the development of novel immunoprevention/immuno-interception strategies. International cancer foundations have also started recognizing immunoprevention and immune interception with the American Association for Cancer Research, Cancer Research UK and the Society for Immunotherapy of Cancer each implementing programming focused on this area. This review will present recent advances, opportunities, and challenges in the emerging field of cancer immune prevention and immune interception.

Interleukin 2 (IL-2) plays a crucial role in T cell growth and survival, enhancing the cytotoxic activity of natural killer and cytotoxic T cells and thus functioning as a versatile master proinflammatory anticancer cytokine. The FDA has approved IL-2 cytokine therapy for the treatment of metastatic melanoma and metastatic renal cell carcinoma. However, IL-2 therapy has significant constraints, including a short serum half-life, low tumor accumulation, and life-threatening toxicities associated with high doses. Oncolytic viruses (OVs) offer a promising option for cancer immunotherapy, selectively targeting and destroying cancer cells while sparing healthy cells. Numerous studies have demonstrated the successful delivery of IL-2 to the tumor microenvironment without compromising safety using OVs such as vaccinia, Sendai, parvo, Newcastle disease, tanapox, and adenoviruses. Additionally, by engineering OVs to coexpress IL-2 with other anticancer transgenes, the immune properties of IL-2 can be further enhanced. Preclinical and clinical studies have shown promising antitumor effects of IL-2-expressing viral vectors, either alone or in combination with other anticancer therapies. This review summarizes the therapeutic potential of IL-2-expressing viral vectors and their antitumor mechanisms of action.