A regulatory pathway involving retinoic acid and calcineurin demarcates and maintains joint cells and osteoblasts in regenerating fin.

During zebrafish fin regeneration, blastema cells lining the epidermis differentiate into osteoblasts and joint cells to reconstruct the segmented bony rays. We show that osteoblasts and joint cells originate from a common cell lineage, but are committed to different cell fates. Pre-osteoblasts expressing runx2a/b commit to the osteoblast lineage upon expressing sp7, whereas the strong upregulation of hoxa13a correlates with a commitment to a joint cell type. In the distal regenerate, hoxa13a, evx1 and pthlha are sequentially upregulated at regular intervals to define the newly identified presumptive joint cells. Presumptive joint cells mature into joint-forming cells, a distinct cell cluster that maintains the expression of these factors. Analysis of evx1 null mutants reveals that evx1 is acting upstream of pthlha and downstream of or in parallel with hoxa13a Calcineurin activity, potentially through the inhibition of retinoic acid signaling, regulates evx1, pthlha and hoxa13a expression during joint formation. Furthermore, retinoic acid treatment induces osteoblast differentiation in mature joint cells, leading to ectopic bone deposition in joint regions. Overall, our data reveal a novel regulatory pathway essential for joint formation in the regenerating fin.

GAS2L3, a target gene of the DREAM complex, is required for proper cytokinesis and genomic stability.

The mammalian DREAM complex is a key regulator of cell-cycle-regulated gene transcription and drives the expression of many gene products required for mitosis and cytokinesis. In this study, we characterized GAS2L3, which belongs to the GAS2 family of proteins with putative actin- and microtubule-binding domains as a target gene of DREAM. We found that GAS2L3 localizes to the spindle midzone and the midbody during anaphase and cytokinesis, respectively. Biochemical studies show that GAS2L3 binds to and bundles microtubules as well as F-actin in vitro. Strikingly, the RNAi-mediated knockdown of GAS2L3 results in chromosome segregation defects in multinucleated cells and in cells with multi-lobed nuclei. Likewise, chronic downregulation of GAS2L3 causes chromosome loss and aneuploidy. Time-lapse videomicroscopy experiments in GAS2L3-knockdown cells reveal abnormal oscillation of chromatin and the spindle during cytokinesis. Taken together, our data reveal novel, important roles of GAS2L3 for faithful cell division. Our work thus contributes to the understanding of how DREAM regulates cytokinesis.

Convenient Access to Expert-Reviewed Health Information via an Alexa Voice Assistant Skill for Patients With Multiple Myeloma: Development Study.

BACKGROUND: Patients with multiple myeloma (MM) have high information needs due to the complexity of the disease and variety of treatments. Digital voice assistants provide support in daily life and can be a convenient tool that even older patients can use to access health information. Voice assistants may therefore be useful in providing digital health services to meet the information needs of patients with MM. OBJECTIVE: We aim to describe and report on the development, content, and functionality of the first Amazon Alexa voice assistant skill for patients with MM in Germany with the goal of empowering and educating patients. Further, we share data on skill usage and first learnings. METHODS: In a cocreation workshop with MM patient organizations and MM medical experts in Germany, Takeda Oncology discussed the development and content of the Alexa skill Multiple Myeloma. Patient information on MM disease, diagnostics, and therapy was presented in a question-and-answer format, reviewed by experts, and programmed into the skill. Additionally, a search function for finding patient support groups within a perimeter of 200 km around the users and a myeloma quiz functionality with multiple-choice questions were integrated into the skill. Aggregated retrospective data on the total number of skill installations and skill usage were retrieved from an Amazon Alexa developer account, and a web-based patient survey was conducted on the Takeda Oncology website. RESULTS: The Alexa skill Multiple Myeloma was launched in September 2019. It was available free of charge on the German Amazon Alexa skill store between September 2019 and March 2022 and could be used with devices featuring the Amazon Alexa voice assistant. Since the launch in September 2019 and up to July 2021, a total of 141 users have installed the skill. Between July 2020 and July 2021, a total of 189 skill sessions with 797 utterances were analyzed. The most popular inquiries were searches for patient support groups near the users (58/797, 7.3%), followed by inquiries about information on MM disease (53/797, 6.6%) and the quiz (43/797, 5.4%). The web-based survey on voice assistant usage and the feedback on the Alexa skill Multiple Myeloma were collected from 24 participants and showed that 46% (11/24) of participants would recommend the Alexa skill. Nonusers of voice assistants (11/24, 46%) stated that data protection concerns (7/11, 64%) and a lack of need (6/11, 55%) were the most important factors of not using voice assistants. CONCLUSIONS: The Alexa skill Multiple Myeloma offers patient-friendly and expert-reviewed answers and explanations for medical terms related to MM disease, diagnostics, and therapy, as well as connections to patient support groups and a quiz functionality. In the future, the skill can be extended with new content and functionalities, such as medication adherence support.

Treatment of relapsed refractory multiple myeloma: which new PI-based combination treatments do patients prefer?

BACKGROUND AND OBJECTIVES: This study describes preferences of German relapsed refractory multiple myeloma (RRMM) patients with novel proteasome inhibitor-based combination treatments. METHODS: Patients with a minimum age of 18 years and a diagnosis of RRMM were included. Their preferences were assessed using a discrete choice experiment design, which was developed based on a literature review and two patient focus group discussions. The final discrete choice experiment design consisted of four attributes, namely "therapy application regimen," "time without progression of disease," "possibility of grade ≥3 adverse events (AEs) affecting the blood," and "possibility of grade ≥3 AE heart failure." RESULTS: Analysis was based on 84 patients (36.9% females, mean age 62.7 years, mean multiple myeloma disease duration 5.5 years). Among the tested attributes, "therapy application regimen" was assigned the highest importance for treatment decisions (38.8%), the second important attribute was "time without progression of disease" (38.7%), followed by "possibility of AE heart failure" (13.9%) and "possibility of AEs affecting the blood" (8.6%). Patients preferred oral intake once a day and once a week over other application modes such as oral intake once a day and once a week plus twice-weekly infusions. Furthermore, they preferred longer disease progression-free time and lower risk of grade ≥3 AEs. The highest overall utility was derived for ixazomib + lenalidomide + dexamethasone (utility: 3.218), compared with lenalidomide + dexamethasone (2.769), and carfilzomib + lenalidomide + dexamethasone (1.928). CONCLUSION: RRMM patients prefer treatments with an all-oral application, a longer disease-progression-free time, and a lower probability of AEs. If patients face tradeoffs, they accept a lower progression-free time and/or higher AE rates to get an all-oral therapy.

Lin9, a subunit of the mammalian DREAM complex, is essential for embryonic development, for survival of adult mice, and for tumor suppression.

The retinoblastoma tumor suppressor protein (pRB) and related p107 and p130 "pocket proteins" function together with the E2F transcription factors to repress gene expression during the cell cycle and development. Recent biochemical studies have identified the multisubunit DREAM pocket protein complexes in Drosophila melanogaster and Caenorhabditis elegans in regulating developmental gene repression. Although a conserved DREAM complex has also been identified in mammalian cells, its physiological function in vivo has not been determined. Here we addressed this question by targeting Lin9, a conserved core subunit of DREAM. We found that LIN9 is essential for early embryonic development and for viability of adult mice. Loss of Lin9 abolishes proliferation and leads to multiple defects in mitosis and cytokinesis because of its requirement for the expression of a large set of mitotic genes, such as Plk1, Aurora A, and Kif20a. While Lin9 heterozygous mice are healthy and normal, they are more susceptible to lung tumorigenesis induced by oncogenic c-Raf than wild-type mice. Together these experiments provide the first direct genetic evidence for the role of LIN9 in development and mitotic gene regulation and they suggest that it may function as a haploinsufficient tumor suppressor.