License Fees Research Articles

Brighter horizons: The necessity of concentrated sponsorship targeted toward minoritized student pharmacists

Due to the effects of structural racism, disproportionately lower numbers of Black, Hispanic or LatinX, American Indian, and Alaska Native students pursue a career in pharmacy and successfully matriculate into the profession. Despite these disparities being present for many years, little progress has been achieved in diversifying the pharmacy profession, resulting in a persistent lack of diversity within pharmacy leadership across employers and pharmacy organizations. Consistent with recent recommendations for improving diversity in pharmacy, the PharmGradWishlist (PGWL) initiative was created as a way for practicing pharmacists and organizations to provide direct financial sponsorship to racially and ethnically minoritized trainees to offset costs incurred during training and during the transition from student to practicing pharmacist. Many of these costs, such as residency and fellowship application fees, job interview travel costs, board exam and licensing fees, and moving expenses, are not typically subsidized by federal student funding. Offsetting these costs is an important way to reduce barriers to entering the profession and postgraduate training, the latter of which may be particularly important in trainees’ pursuit of academic and leadership positions in pharmacy. The initial development and advertisement of the initiative occurred through social media and the grassroots efforts of the PGWL team, a group of 10 volunteer pharmacists from across the country, and resulted in generous donations from a small proportion of practicing pharmacists nationwide. It is now time for the profession as a whole to embrace the role of direct sponsorship in improving diversity in the profession. We call upon pharmacists and pharmacy organizations to advocate for and participate in financial sponsorship of racially and ethnically minoritized trainees and pharmacists as a way to increase diversity and promote health equity.

Public Participation and Public Finance Management Performance in Embu County Government, Kenya

The resource absorption rates at the county depicts a situation where all counties did not meet their local revenue target in the fiscal year 2015/16. On average, all the 47 counties achieved 69.3% of their targeted local revenue collection; 19 out of 47 counties (40%), achieved more than 70% of their revenue target in Financial Year2015/16. This is evidenced by governors being summoned at Senate and others who have been impeached due to violation of the Public Finance Management Act 2013, and violation of the County Governments Act 2012. The general objective of the study was to determine the effect of public participation on public finance management in Embu County Government, Kenya. The specific objectives were to examine the influence of communication and access to information, capacity of the public to participate and active involvement on public finance management in Embu County Government, Kenya. The study was guided by five theories including Democratic Theory, Stakeholder Theory, stewardship theory, ladder of participation and Agency Theory. The study adopted an explorative research design. The unit of observation was Embu County government, and the unit of analysis was a total 12 participants consisting of 1 chief, 1 assistant chief, 5 county officials and 5 village elders from each sub county who are involved in the public participation forum in Embu County. Data was collected using questionnaires and analyzed using descriptive and inferential statistics. The data was then presented in tables and figures. The study found out that the extent at which the identified projects implemented in Embu County was at very large extent, the extent at which the Embu County collect and account for the budgeted internal revenues e.g. parking fees, rates, license fees followed with large extent, the extent at which the public participation influence public financial management performance in Embu County was moderate, the extent at which the county officials and employees were engaged without following the laid down procedures was least extent. The study also found out that the respondents indicated that the Embu County government use the print media to invite the public-to-public participation forums, Embu County government use the social media (WhatsApp, Facebook and Tweeter) to invite the public-to-public participation forums, the Embu County government use the print media (newspapers and displays at public places) to invite the public-to-public participation forums, the Embu County government use telephone calls (mobile and landline) to invite the public-to-public participation forums. The study further found out that the public participation leader gives everyone a chance to participate during public participation forums, respondents indicated that they consider their profession/education sufficient for effective public participation, the questions asked and responded to during the public forums were satisfactory.

Structural Changes in Radio and Impacts on Music Listeners

The current study explores structural changes in radio and assesses their impact on music listeners in Pakistan utilizing the theoretical underpinnings of the political economy of communication. A mixed-method approach was used to answer research questions and evaluate hypotheses. A contextual analysis was utilized to evaluate the qualitative analysis after the interviews with 14 experts in the Pakistani radio industry were transcribed. Cross-tabulation with chi-square test, Independent Sample T-test, and AVOVA with Tukey tests were used to analyze survey responses from 200 respondents in order to determine the correlations between variables. The findings show that structural trends in the radio industry's deregulation and commercialization have curtailed access to a varied range of listeners in Pakistan. According to the research, deregulation, licensing fees, new license fees, and the restriction of Indian music, revenue generation, and licensing to nonprofit FM radio have all limited access to a diverse spectrum of radio music listeners in Pakistan. This trend also leads broadcast media in Pakistan to produce and disseminate more entertainment programming rather than information and education programs.

Oil Revenue and Economic Growth of Nigeria: 1990 – 2019

The oil and gas business in Nigeria has brought unparalleled fluctuations to the Nigerian economy, mostly in the past five decades after it substituted agriculture as the basis of the Nigerian economy. This study investigated the relationship between oil revenue and economic growth in Nigeria. It spanned through the period 1990 through 2019. The specific objectives are to investigate the relationship between crude oil/gas export, petroleum profit tax/royalty, domestic crude oil sales, oil licensing fees on real gross domestic product and real gross national product in Nigeria. And also, ascertain whether the exchange rate moderates the relationship between oil revenue and economic growth in Nigeria. The study employed an ex post facto research design and the secondary data used for the investigation were sourced from the Central Bank of Nigeria (CBN) statistical bulletin, Federal Inland Revenue Service Fact Book and the World Bank Development Website. Descriptive Statistics, Pearson Moment Correlation Coefficient and Ordinary Least Square Multiple Regression Statistical tools were used in the study. The results revealed that Crude oil/gas export has a significant and negative relationship with the real gross domestic product in Nigeria; Petroleum profit tax/royalty has a significant and positive relationship with real gross domestic in Nigeria; Domestic crude oil sales have an insignificant and negative relationship with real gross domestic product in Nigeria; Oil licensing fees have an insignificant and negative relationship with real gross domestic product in Nigeria; The study concluded that there is a significant relationship between oil revenue and economic growth in Nigeria. The study recommends that the government should effectively and efficiently utilize the oil fund in strategic development projects so as to reduce the rate of poverty and facilitate economic growth.

Consumer welfare, licensing, and exclusive dealing with vertically- and horizontally-differentiated products

We explore the existence of anticompetitive exclusive dealing when the downstream firm produces both vertically-and horizontally products. Our study shows that the licensing scheme has crucial ramifications for considering the possibility of exclusive dealing. For royalty licensing, the less the horizontal differentiation or the similar the quality of the two products is, the less possibility there is for exclusive dealing. However, under two-part tariff licensing, a pure fixed fee is preferred by a licensor, it is more likely to foster exclusive dealing. Moreover, we explore the consumer welfare implications and find that royalty licensing may prevent anticompetitive exclusion and lead to greater consumer surplus, which runs against the conventional wisdom that fixed fee licensing is socially desirable. In addition, royalty licensing can be favorable to the outside licensor if the licensor can commit at the beginning of the game.

Sunk Cost Analysis of Frequency Spectrum Auction Prices in Indonesia Using Data Envelopment Analysis

The purpose of this study is to investigate the sunk cost argument on the auction price of the frequency spectrum by measuring the efficiency of the frequency license fee paid by the auction winner. The efficiency method employs a nonparametric approach, namely Data Envelopment Analysis (DEA) with the Linear Programming method, with the hypothesis model tested using Path Analysis. The dataset under consideration was Indonesia's Big Three Service Providers (2015-2020). The results show that the spectrum auction price is efficient in its impact on service prices, revenues, and income taxes. The spectrum auction price has no effect, lending credence to the argument that the spectrum auction price is considered a sunk cost in the telecommunications industry. The research reason is that the higher the auction price, combined with the lower cost of service due to high competition, the harder it is for small businesses to win the auction. The winning bidder will remain under the control of the prominent organizers, so the government issues a merger policy for small businesses to compete in spectrum auction bids.

Cross-State Licensure Laws for Telehealth Evolve During Pandemic

Cross-State Licensure Laws for Telehealth Evolve During Pandemic

Bunching with the Stars: How Firms Respond to Environmental Certification

Voluntary environmental certification programs have been a popular tool used by governments, industry groups, and nonprofit organizations alike. A central question in the design of such programs is who should pay for them. In a context where firms respond strategically to a certification, the answer to this question is a priori ambiguous and, ultimately, empirical. This paper provides important insights on this question using ENERGY STAR, a voluntary certification program for energy-efficient products, as a case study. I show that firms are highly strategic with respect to this certification and extract consumer surplus associated with certified products via three mechanisms. They offer products that bunch at the certification requirement, differentiate certified products in the energy and nonenergy dimensions, and charge a price premium on certified products. I use these findings to motivate a structural econometric model with firms’ strategic behaviors with respect to product line and pricing decisions and to investigate the incidence of a certification licensing fee to fund the certification program. This paper was accepted by Juanjuan Zhang, marketing.

Product Design for Recycling Under Different Licensing Fee Schemes

Product Design for Recycling Under Different Licensing Fee Schemes

К вопросу об особенностях гражданско-правовой ответственности за незаконное использование изображений персонажей художественных произведений как объектов авторских прав

Theoretical and practical issues of civil liability of business entities that use images of fictional characters, mainly from feature films, without having the appropriate permission from the copyright holders, have been studied. The author has established that the illegal use of images of characters, as a rule, is associated with the production of goods whose packaging or appearance embody well-known fictional images. Consumers are subconsciously guided by these images and form demand necessary to increase sales. The infringer, embodying the images of famous characters protected both by copyright and as part of trademarks in goods and on their packaging, significantly reduces their costs due to non-payment of a license fee to the right holder and thereby reduces prices for these goods. The actual damage may be the costs of the right holder, which they have incurred or will inevitably incur when restoring their violated right. These costs can be incurred when making test purchases of counterfeit goods on which cartoon characters are placed, upon receipt of information fixing relevant violations, information on sales volumes of counterfeit goods, etc. Losses caused by violation of the exclusive right to depict a fictional character may also be expressed in the form of lost profits. Such a loss may manifest itself either in the failure of the right holder to receive remuneration for its illegal use, or in the failure to receive income from the independent use of the character in the implementation of certain entrepreneurial activities. If it is proved that the illegal user received income in a certain amount as a result of such a violation, then the right holder has the right to demand compensation, along with other losses and lost profits, in an amount not less than the income received by the violator. From the analysis of the judicial practice widely presented in this article, it follows that the most common way to protect violated exclusive rights to images of fictional characters is to claim monetary compensation, which is a borrowing from the Anglo-American legal order with its institution of statutory damages. The article analyzes the judicial practice related to the application of this sanction, notes its main shortcomings and concludes that the explanations of the Supreme Court of the Russian Federation in this part are rather superficial, not presented systematically and contradict the doctrinal provisions of the science of civil law. The author concludes that, to eliminate this shortcoming and to implement the principles of justice, equality of participants in civil relations, it is necessary to exclude the lower threshold of compensation from these norms.

A Study on the International Competitiveness of Korea and China in Service Trade

Purpose This study compares and analyzes the trends in service trade between Korea and China by analyzing the RCA index, SRCA index, and TSI index. Design/Methodology/Approach For this analysis using the UN Comtrade DB, the service trade in 11 sectors was analyzed using Extended Balance of Payments Services (EBOPS) 2002 between Korea and China for 8 years (2010 to 2018). The comparative advantage was measured by calculating the present comparative advantage index (RCA), the revealed symmetric comparative advantage (SRCA), and the trade specialization index (TSI). Then, the level of competition was measured by analyzing the export competition (ESI) of the service sectors between Korea and China. Findings In Korea and China, the transportation and construction sectors, which are highly related to manufacturing, have relatively high international competitiveness; however, international competitiveness is weakening in other service trade sectors. Research Implications In the case of royalties and license fees, if international competitiveness is weakened, as product exports increase, the size of the service deficit in that sector will inevitably increase. Accordingly, it should be human investment related to infrastructure construction and human education, such as royalties and license fees.

Are the compensations given to mining enterprises due to the overlapping with other investments in Turkey enough? Expectations of compensation for profit deprivation

Generally, there can be very high differences among the alternative investment costs of the other activity which overlaps with mining. This situation can oblige the overlapping of the alternative investments of these activities that overlap with mining in the available location with mining sites due to not being able to be preferred. There is no mistake in shutting down mining activities completely or partially in these kinds of investment overlaps. However, are mining operations, whose activities have been finalized due to not being foreseen beforehand and who experience investment loss, given compensation for profit deprivation and investment losses? Are the investment costs of mining operations covered precisely? In these kinds of investment overlaps in Turkey, particularly license and land use fees of mining enterprises are not paid, compensation for profit deprivation are not paid by considering proved mineral reserve, and investment costs are given only to a limited extent. This situation occurred e.g. in Izmir as a result of the overlapping of 15 mining operations with a highway investment project route. The expectation of mining enterprises in Turkey and the opinion of doctrine considering Turkish Constitution and legislative provisions are that all mining investment losses to met completely and compensation for profit deprivation to met to a certain extent. In the answers given to the question asked for mining enterprises through SurveyMonkey program in 2018, most of the enterprises request that compensation for lost profit is given from the proved reserved to a certain extent due to mineral reserve losses. Along with the available investment costs foreseen in Turkish Mining Regulation, several times more of six-month operating costs determined as profit deprivation in nationalizations can be given to all mining enterprises whose activities are terminated in these investment overlaps. Or, a different method can be applied in the calculation of compensation for profit deprivation. In this method, it is suggested that a new calculation for compensation for profit deprivation shall be made by considering pit sale prices and proved reserves. There are distinctions made as the operations -whose proved reserves are determined according to UMREK, The operations whose proved reserves are not determined according to UMREK (or whose proved reserves determined in mining operating projects), “mining enterprise whose proved mineral reserve is determined according to UMREK (congruent with CRIRSCO), or not determined according to UMREK (considered mining operating project in Turkey), “open-pit or underground mining operation” and “mining operation providing or not providing additional added value by processing the minerals in their country and facilities.” Considering these differences, it was suggested to apply different compensation rates.

The Decision-Making Analysis on End-of-Life Vehicle Recycling Extending Producer Responsibility Policy in the Context of Remanufacturing

Abstract For ensuring an ideal recycling rate, the extending producer responsibility (Extending Producer Responsibility) has been recognized by more and more governments. And an increasing number of original vehicle manufacturers (Original Vehicle Manufacturers) participated in their End-of-Life vehicles (End-of-Life Vehicles) reusing to fulfil their social responsibility. The Original Vehicle Manufacturers can do well in auto parts remanufacturing due to their knowledge and skills of vehicle manufacturing, and the third-party vehicle recyclers are more capable of recycling the End-of-Life Vehicles due to their skills and facilities of waste recycling. To better improve the End-of-Life Vehicle reusing businesses and improve the efficiency and quality of End-of-Life Vehicle recycling and remanufacturing, this research built a remanufacturing system with dual-cycle End-of-Life Vehicle recycling. Moreover, for the well operation of this system, the researchers established the game model between Original Vehicle Manufacturers and third-party recyclers with the government involved and analyzed this model with evolutionary game theory. This research found that there are two evolutionary stable strategies (ESS) for the two groups. For well operation of remanufacturing system with dual-cycle End-of-Life Vehicle recycling, the government need to use the Extending Producer Responsibility policy to impose strict restrictions on Original Vehicle Manufacturers doing End-of-Life Vehicle recycling through promoting Original Vehicle Manufacturers’ recycling quantity standard and the penalty for insufficient End-of-Life Vehicle recycling. Further, this research also found that the licensing fee that the third-party recycler needs to pay the Original Vehicle Manufacturer for getting the recycling business plays a vital role in the ESS.

4O Nivolumab (NIVO) + ipilimumab (IPI) versus chemotherapy (chemo) as first-line (1L) treatment for advanced NSCLC (aNSCLC) in CheckMate 227 part 1: Efficacy by KRAS, STK11, and KEAP1 mutation status

4O Nivolumab (NIVO) + ipilimumab (IPI) versus chemotherapy (chemo) as first-line (1L) treatment for advanced NSCLC (aNSCLC) in CheckMate 227 part 1: Efficacy by KRAS, STK11, and KEAP1 mutation status

122MO Nivolumab (NIVO) + ipilimumab (IPI) as first-line (1L) treatment (tx) for patients (pts) with advanced NSCLC (aNSCLC) and baseline (BL) brain metastases (mets): Intracranial and systemic outcomes from CheckMate 227 Part 1

122MO Nivolumab (NIVO) + ipilimumab (IPI) as first-line (1L) treatment (tx) for patients (pts) with advanced NSCLC (aNSCLC) and baseline (BL) brain metastases (mets): Intracranial and systemic outcomes from CheckMate 227 Part 1

The Evolutionary Game Analysis and Optimization Algorithm of Electric Vehicle Cell Innovation Diffusion Based on a Patent Pool Strategy

A patent pool strategy was proposed for use in the electric vehicle cell industry to manage patent licensing disputes and litigation. How to promote EV cell innovation diffusion under a patent pool scenario is unclear. We introduced an innovation diffusion channel model comprising different players with patent licensing relationships and market competition relationships following evolutionary game analysis and simulation. We found the interlinked factors that influenced evolutionary stable strategies with a sensitivity test on all factors to identify the important and unimportant factors. To achieve the maximum return for the players, an optimization algorithm was introduced to find the maximum weighted object function. The decision and policy makers could focus on important factors such as improving the technology’s competitive advantages, delivering more profits to its licensees with reasonable licensing fees, and finding the best patent pool strategy with the support of the optimization algorithm to balance the competition relationships and patent licensing relationships between players.

Rentier capitalism in question

Rentier capitalism is an idea that has been drawing increasing attention. This article identifies the incoherence of the idea and then addresses its weaknesses as a critique of contemporary political economy. In the first place, it derives from a view of capitalism which privileges exchange rather than production. This explains its view of the relation between the monopoly control of assets and competition. While there may indeed be a current tendency to invest in assets, that does not necessarily act as a deadweight on production; rather it can act as an incentive to rationalize the labor process with a view to increasing the extraction of surplus value. It is suggested that one of the ways in which industrial capital accommodates itself to increasing rents, license fees and the like, is through a turn to extracting value in its absolute form; which may in then shed light on the stagnation of wage levels that has accompanied the turn to investing in assets.

Health insurance coverage and switching among people with hemophilia A in the United States

BACKGROUND: Hemophilia A (HA) is marked by substantial economic burden, including costs of ongoing treatment, increased monitoring, bleed events, and other health care utilization associated with managing the disease and comorbidities related to the disease. Gene therapies and other anticipated breakthrough treatments hold potential to substantially offset long-term traditional factor VIII (FVIII) prophylaxis in specific populations. Fragmentation of the US insurance system, however, may impact payers' approaches to coverage of new treatments, given concerns about patients "switching" insurance and the payer's ability to offset costs over time. OBJECTIVE: To assess insurance coverage and switching across payers among people with severe HA (SHA) using real-world data. METHODS: Adult men with SHA (FVIII measuring < 1%) in the American Thrombosis and Hemostasis Network dataset between January 2013 and September 2019 were identified. Patients' primary insurance category (ie, commercial, Medicaid, Medicare) and insurance switching over time were described. Outcomes included distribution of current primary insurance coverage by category and mean years of coverage per payer for commercially insured patients, including those with 2 or more commercial payers, and for those who switched insurance categories (eg, coverage by a commercial payer and government payer). RESULTS: Among the cohort of patients with SHA (N = 3,677), 51.9% had commercial primary insurance and 29.0% had coverage by Medicaid (including state-funded programs). The mean duration of follow-up in the database was 6.3 years for patients with at least 1 year of follow-up. Among patients who had ever been commercially insured, 74.9% had the same commercial payer for the entire follow-up period. The mean time covered by the same commercial insurance was 4.8 years. Only 7.5% of patients switched insurance categories (eg, from commercial to Medicaid). Among those who switched categories, patients averaged 3.9 years of commercial coverage, 4.0 years of Medicaid coverage, and 4.8 years of Medicare coverage during the follow-up period. CONCLUSIONS: Both commercially and government-insured patients with SHA typically maintain continuous coverage for extended periods, with limited switching between payers and insurance categories over time. These findings suggest that should breakthrough treatments be approved, payers would likely be able to realize substantial cost savings associated with avoiding long-term prophylactic therapies during the several years after treatment. DISCLOSURES: This study was funded by BioMarin Pharmaceutical Inc. Hinds, Chen, and Sammon are employees of BioMarin Pharmaceutical Inc. and own stock/stock options. Solari was an employee of BioMarin Pharmaceutical Inc. at the time of the study. Pezalla is CEO of Enlightenment Bioconsult, LLC. He, Cheng, and Recht are, or were at the time of this study, employees of American Thrombosis and Hemostasis Network (ATHN), which has received ATHNdataset licensing and other fees from BioMarin Pharmaceutical Inc. Research funding to Recht's employers has come from Bayer, BioMarin Pharmaceutical Inc., CSL Behring, Genentech, Grifols, Hema Biologics, LFB, Novo Nordisk, Octapharma, Pfizer, Sanofi, Spark, Takeda, and uniQure. Recht has also worked as a consultant for Catalyst Biosciences, CSL Behring, Genentech, Hema Biologics, Kedrion, Novo Nordisk, Pfizer, Sanofi, Takeda, and uniQure; sits on the board of directors of the Foundation for Women and Girls with Blood Disorders and of Partners in Bleeding Disorders; and is an employee of the Oregon Health & Science University. Data from this study were presented as a poster at AMCP Nexus 2021; October 18-21, 2021; Denver, CO.

Eomes and T-Bet Expression Are Required By Mature Primary Human NK Cells for Anti-Leukemia Responses In Vivo

Natural Killer (NK) cells are innate lymphoid cells that respond to hematologic cancers via cytotoxicity (perforin/granzyme and death receptors) and cytokine/chemokine production, yet the molecular determinants underlying their proliferation, function, and persistence are poorly understood. There are promising reports of pre-clinical and clinical NK cell responses to leukemia and lymphoma, which represent a nascent cellular therapy for these blood cancers. The T-box transcription factors (TFs) Eomes and T-bet are expressed by NK cells throughout their lifespan, and are required for development as evidenced by NK cell loss in Eomes and T-bet deficient mice. However, the roles of these TFs in mature human NK cell molecular programs and functions remain unclear.We hypothesized Eomes and T-bet, which are the only T-box TFs expressed in NK cells, are critical regulators of NK cell homeostasis and functionality, and are necessary for proper mature NK cell responses. To address this, we utilized the CRISPR-Cas9 system to genetically delete both Eomes and T-bet in primary human NK cells isolated from healthy donors, and investigated their role beyond guiding NK cell development, specifically in the anti-leukemia response. Gene-editing of primary human NK cells has been technically challenging, thus most reports that modified NK cells were performed with cell lines, in vitro-differentiated, or highly expanded NK cells that likely do not reflect primary human NK cell biology.Here, we introduced Cas9 mRNA and sgRNA targeting T-bet and Eomes by electroporation into unexpanded primary human NK cells isolated from healthy donors using the MaxCyte GT system. We observed highly efficient reductions of Eomes and T-bet protein expression, quantified by flow cytometry (p < 0.0001, Fig A-B) without viability differences between control (sgRNA targeting TRAC, an unexpressed locus in NK cells), and Eomes/T-bet double CRISPR-edited (DKO) cells after one week in vitro. To study Eomes and T-bet in NK cell anti-leukemia response, control or DKO primary human NK cells were engrafted into NSG mice, supported with human IL-15, and challenged with K562 leukemia cells. Utilizing bioluminescent imaging to visualize leukemia burden, we observed that NK cells lacking both TFs were unable to suppress leukemia growth in vivo.To understand the mechanism responsible for impaired leukemia control, we investigated in vivo persistence and proliferation, cytotoxic effector molecule expression, as well as ex vivo degranulation and cytokine production of DKO NK cells compared to control NK cells. DKO or control human NK cells were transferred into NSG mice and supported with human IL-15. After 2-3 weeks, significantly fewer (<30%) DKO NK cells persisted compared to control NK cells: spleen (5-fold decrease, control 240e3±65e3 vs DKO 47e3±15e3 NK cells, p<0.01, Figure C), blood (6-fold decrease, p<0.01), and liver (4-fold decrease, p<0.05). Using intracellular flow cytometry, double T-bet/Eomes CRISPR-edited NK cells that lacked both Eomes and T-bet protein after in vivo transfer were identified. A proliferative defect was evident in flow-gated DKO (62±6% undivided), compared to unedited (WT) NK cells (4±2% undivided) assessed by CellTrace Violet dilution (Figure D). In addition, there were marked reductions in granzyme B and perforin protein (p<0.001) in flow-gated DKO NK cells compared to controls. To assess DKO NK cell functional capacity, we performed an ex vivo functional assay on NK cells from spleens of the NSG mice as effectors, and K562 targets or IL-12/15/18 stimulation for 6 hours. Degranulation to K562 targets was impaired (p<0.05), and IFN-γ production was reduced (p<0.0001) after cytokine stimulation in flow-gated DKO NK cells (Figure E).Thus, CRISPR-editing of unexpanded, primary human NK cells revealed that Eomes and T-bet are required by mature human NK cells for their function and homeostasis, distinct from their role in development. This is translationally relevant, as defects in proliferation and function of human DKO NK cells manifested markedly reduced response against human leukemia cells in vivo in xenografts. These findings expand our understanding of key molecular regulators of mature NK cell homeostasis and function, with the potential to provide new avenues to enhance NK cell therapy. [Display omitted] DisclosuresBerrien-Elliott: Wugen: Consultancy, Patents & Royalties: 017001-PRO1, Research Funding. Foltz-Stringfellow: Kiadis: Patents & Royalties: TGFbeta expanded NK cells; EMD Millipore: Other: canine antibody licensing fees. Fehniger: HCW Biologics: Research Funding; Compass Therapeutics: Research Funding; Affimed: Research Funding; ImmunityBio: Research Funding; Wugen: Consultancy, Current equity holder in publicly-traded company, Patents & Royalties: related to memory like NK cells, Research Funding; Kiadis: Other; OrcaBio: Other; Indapta: Other.

Cytokine-Induced Memory-like NK Cells Have a Distinct Single Cell Transcriptional Profile and Persist for Months in Adult and Pediatric Leukemia Patients after Adoptive Transfer

Natural killer (NK) cells are innate lymphoid cells that mediate anti-tumor responses and exhibit innate memory following stimulation with IL-12, IL-15, and IL-18, thereby differentiating into cytokine-induced memory-like (ML) NK cells. ML NK cells have well-described enhanced anti-tumor properties; however, the molecular mechanisms underlying their enhanced functionality are not well-understood. Initial reports of allogeneic donor ML NK cellular therapy for relapsed/refractory (rel/ref) acute myeloid leukemia (AML) demonstrated safety and a 47% CR/CRi rate (PMID32826231). In this setting, allogeneic ML NK cells are rejected after 3 weeks by recipient T cells, which precludes long-term evaluation of their biology. To address this limitation, we conducted a clinical trial for rel/ref AML patients that added adoptive transfer of same-donor ML NK cells on day +7 of a reduced-intensity conditioning (RIC) MHC-haploidentical HCT, followed by 4 doses of IL-15 (N-803) over 2 weeks (NCT02782546). Since the ML NK cells are from the HCT donor, they are not rejected, but remain MHC-haploidentical to the patient leukemia. Using samples from these patients, we profiled the single cell transcriptomes of NK cells using multidimensional CITE-seq, combining scRNAseq with a custom NK panel of antibodies.To identify donor ML NK cells in an unbiased fashion, we developed a CITE-seq ML NK classifier from in vitro differentiated paired conventional NK (cNK) and ML NK cells. This classifier was applied via transfer learning to CITE-seq analyzed samples from the donor (cNK cells) and patients at days +28 and +60. This approach identified 28-40% of NK cells as ML at Day +28 post-HCT. Only 1-6% of donor peripheral blood NK cells and 4-7% of NK cells in comparator leukemia patients at day +28 after conventional haplo-HCT alone were identified as ML NK cells (Fig 1A). These ML NK cells had a cell surface receptor profile analogous to a previously reported mass cytometry phenotype. Within the CITE-seq data, ML NK cells expressed a transcriptional profile consistent with enhanced functionality (GZMK, GZMA, GNLY), secreted proteins (LTB, CKLF), a distinct adhesome, and evidence of prior activation (MHC Class II and interferon-inducible genes). ML NK cells had a unique NK receptor repertoire including increased KIR2DL4, KLRC1(NKG2A), CD300A, NCAM1(CD56) , and CD2 with decreased expression of the inhibitory receptor KLRB1(CD161). Furthermore, ML NK cells upregulated HOPX, a transcription factor implicated in memory T cells and murine CMV adaptive NK cells. Additionally, ML NK cells downregulated transcription factors related to terminal maturation (ZEB2) and exhaustion (NR4A2).We next sought to identify changes during ML differentiation in patients post-HCT from day +28 to +60 post-HCT. Trajectory analysis identified a ML NK cell state distinct from cNK cells that was present at least 60 days post-HCT (Fig 1B). The ML transcriptional phenotype continued to modulate during late differentiation, including downregulation of GZMK and NCAM1, and upregulation of maturation related transcription factors, while maintaining high expression of HOPX. ML NK cells retained their enhanced functionality during in vivo differentiation, as patient ML NK cells had significantly increased IFNγ production compared to cNK cells after restimulation with leukemia targets or cytokines using mass cytometry (Fig. 2). Subsequently, we confirmed the ML CITE-seq profile in an independent clinical trial treating pediatric AML relapsed after allogenic HCT with same-donor ML NK cells (NCT03068819). In this setting, ML NK cells expressed a similar transcriptional signature and persisted for at least 2 months in the absence of exogenous cytokine support. Thus, ML NK cells possess a distinct transcriptional and surface proteomic profile and undergo in vivo differentiation while persisting within patients for at least 2 months. These findings reveal novel and unique aspects of the ML NK cell molecular program, as well as their prolonged functional persistence in vivo in patients, assisting in future clinical trial design. [Display omitted] DisclosuresFoltz: Kiadis: Patents & Royalties: TGFbeta expanded NK cells; EMD Millipore: Other: canine antibody licensing fees. Berrien-Elliott: Wugen: Consultancy, Patents & Royalties: 017001-PRO1, Research Funding. Bednarski: Horizon Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees. Fehniger: Wugen: Consultancy, Current equity holder in publicly-traded company, Patents & Royalties: related to memory like NK cells, Research Funding; ImmunityBio: Research Funding; Kiadis: Other; Affimed: Research Funding; Compass Therapeutics: Research Funding; HCW Biologics: Research Funding; OrcaBio: Other; Indapta: Other.