Identifying Mechanisms Associated with Venetoclax Resistance in Multiple Myeloma (MM)

Venetoclax (VTX) is a selective small-molecule inhibitor of BCL-2 that exhibits antitumoral activity against MM cells presenting lymphoid features and those with translocation t(11;14). Despite its impressive clinical activity, VTX therapy for a prolonged duration can lead to drug resistance. Therefore, it is important to understand the underlying mechanisms of resistance in order to develop strategies to prevent or overcome resistance. In the present study, we established four VTX resistant human myeloma cell lines (HMCLs) from four sensitive HMCLs, including three with t(11;14), in culture with a stepwise increase in treatment dose with VTX. To identify the molecular basis of acquired VTX resistance, whole exon sequencing (WES), mRNA-sequencing (mRNAseq), and protein expression assays were performed in the four isogenic VTX-sensitive/resistant HMCLs and three MM patients with samples collected before VTX administration and after clinical resistance to the drug. Compared with sensitive cell lines and patient samples collected before VTX administration, mRNAseq analysis identified downregulation of BIM and upregulation of BCLXL in both resistant cell lines and MM cells from relapse patients. Other transcriptional changes detected included upregulation of AURKA, BIRC3, BIRC5, and IL32. Enrichment analysis of differentially expressed genes suggested involvement of PI3K and MAPK signaling, likely associated with cytokines, growth factors (EGF, FGF and IGF family members), and receptor tyrosine kinase (EGF and FGF). Western blot analysis was performed to compare BCL2 family expression in resistant cell lines versus sensitive cell lines and it showed upregulation of BCL2 survival members (such as MCL-1 and BCLXL), and downregulation of pro-apoptotic BH3 members (such as BIM and PUMA). BIM expression was completely lost in one resistant cell line, and introduction of exogenous BIM into this cell line enhanced VTX sensitivity. Interestingly, BCL2 was upregulated in some resistant cell lines generated after a long-term treatment with VTX, suggesting BCL2 expression level may not be suitable as a marker of VTX sensitivity for acquired resistance. Unlike in CLL, BCL2 mutations were not identified through WES in any resistant cell lines or primary patient sample harvested after relapse. While 8 genes were mutated in two resistant samples , no clear mutational pattern emerged . Based on the above, we further tested some specific inhibitors in in vitro or ex vivo cell models to help understanding resistant mechanism and identify strategies to overcome VTX resistance. We found that inhibition of MCL-1, with the compound S68345, substantially enhanced VTX sensitivity in three resistant HMCLs and in primary cells from one relapsed MM patient. A BCLXL inhibitor (A155463) only significantly enhanced VTX sensitivity in one resistant cell line after co-treatment with VTX. Co-treatment of the other three resistant cell lines with VTX, S68345 and A155463 resulted in the most synergistic anti-myeloma activity, suggesting those cell lines are co-dependent on MCL-1, BCLXL, and BCL2 for survival, although they are more dependent on MCL-1. We also found that inhibition of PI3K signaling, IGF1, RTK (EGF and FGF) and AURKA significantly increased VTX sensitivity, partially through downregulation of MCL-1, and BCLXL, and upregulation of BIM. Conventional anti-MM drugs such as dexamethasone, bortezomib and lenalidomide, were shown to have little activity on augmenting VTX sensitivity in most resistant cell lines. In summary, we find that acquired resistance to VTX in MM is largely associated with BCL2 family regulation, including upregulation of survival members such as MCL-1, BCLXL, BCL2, and downregulation of pro-apoptotic members, especially BIM. Our study also indicates that upstream signaling involved in BCL2 family regulation during acquired resistance is likely related to cytokine, growth factor, and/or RTK-induced cell signaling such as PI3K. Co-inhibition of MCL-1, or BCLXL, as well as the upstream PI3K, RTK (FGF and EGF), IGF-1 mediated signaling were effective in overcoming VTX resistance. Disclosures Fonseca: Mayo Clinic in Arizona: Current Employment; Amgen: Consultancy; BMS: Consultancy; Celgene: Consultancy; Takeda: Consultancy; Bayer: Consultancy; Janssen: Consultancy; Novartis: Consultancy; Pharmacyclics: Consultancy; Sanofi: Consultancy; Merck: Consultancy; Juno: Consultancy; Kite: Consultancy; Aduro: Consultancy; OncoTracker: Consultancy, Membership on an entity's Board of Directors or advisory committees; GSK: Consultancy; AbbVie: Consultancy; Patent: Prognosticaton of myeloma via FISH: Patents & Royalties; Scientific Advisory Board: Adaptive Biotechnologies: Membership on an entity's Board of Directors or advisory committees; Caris Life Sciences: Membership on an entity's Board of Directors or advisory committees.

Somali society's concept of stewardship: A study from Islamic perspective

This research’s topic revolves around a sensitive and strange issue that usually causes imbalance in family bonding and undermining stewardship in Somali society, which is the issue of leaving maintenance of the family to the women due to lack of supporter and carer for them. Therefore, it makes the highest population of the women in that society engage in different kind of jobs, which gives them the power of controlling homes and the society. The researcher adopts the inductive research methodology to gather pieces of evidence on the custodian of stewardship and how to control the family from the word of Allah and the sayings of the prophet Muhammad, as well as books and articles written by Somali scholars and researchers and United Nation report regarding the women guardianship in Somali society. The researcher also adopts analytical method to analyse the texts gathered from the texts related to the topic, and followed by interview, which is used to collect data related to the topic from nine respondents; four among them are elites, another four are laymen, and the president of (Somali Scholars Association), then analyse the interview and derive the effects of misunderstanding the stewardship on Somali community. In conclusion, the research finds that the civil war is one of the factors contributed to strengthening women stewardship in the Somali community, and that the Somali men misunderstood the true meaning of guardianship due to lack of deep understanding of it. The researcher also observed that addiction of men to the “khat weed” is another factor contributes to the men unseriousness in the Somali community and results to family separation in the community, hence, it leads Somali women to go out for work and get more power over the men, which causes emotional and educational deprive for Somali children, and consequently leads to the behavioral deviation in them. It is also found through the research that lack of state’s security and protection for women, absence of a tangible family regulation, loss of moral supports from religious scholars.

Ending the Family Death Penalty and Building a World We Deserve

U.S. history is rooted in the rationalization of family separation to benefit white supremacy, capitalism and mainstream U.S. values. Because of this dark history, the U.S. history has become the world’s leader of legal destruction of families through termination of parental rights. It is the only country in the world that routinely pays people to adopt children whose parents, often women, very much want to be their parent. The Adoption and Safe Families Act, enacted in 1997, wildly changed the legal landscape of the family regulation system. At that time 47% of the children in the system were Black, and the drug war had been targeting Black men for low level offenses, and labeling Black mothers as “crack moms”. The result was an extreme attack on Black families, for which we have yet to recover. Abolition teaches us to unroot oppressive structures, disrupt and dismantle them while simultaneously supporting a praxis of imagination, healing, and building. In this paper, we encourage people not only to work to repeal ASFA, but to interrogate the imagination which entrenched the legitimacy of ASFA. Part I centers the discussion in our imaginations—the world we want to build, and the demands we are making. Part II moves into a discussion about the counter imagination, the ideas and mythology that created ASFA—the legal framework. In this section, we isolate ASFA as a target for abolition and organizing. Part III moves into a practical discussion about ethical ways to mobilize around ASFA. This section is intended to invite the reader to learn, and question, together. It invites questions, thinking, and problem solving in lieu of providing a recommendation.

The White Supremacy Hydra

Fundamentally, the so-called “child welfare system”—more appropriately named, the family regulation system—is a policing system rooted in white supremacist ideologies and techniques. From its earliest iteration, the family regulation system has functioned to pathologize, control, and punish the families entrapped in its web, most especially Black families. Nevertheless, among many, the myth persists that the family regulation system is one of child protection and family support. This is especially true when discussing the Family First Prevention Services Act of 2018, which—for the first time since the establishment of the modern family regulation system—opens up federal funding streams previously reserved for the removal of children to the foster system to provide prevention services for families in which children have not yet been removed to the foster system. While the Act is a course change in federal family regulation policy, this Article traces how it leaves undisturbed the pathology, control, and punishment central to the policies that preceded it.

Reimagining Schools' Role Outside the Family Regulation System

The United States’ family regulation system often begins with well-intentioned professionals making child protection hotline calls, jeopardizing their own ability to work with families and subjecting the families to surveillance. By the system’s own standards, most of this surveillance leads to no meaningful action. Nowhere is this reality more present than in schools. Educational personnel serve as the leading driver of child maltreatment allegations, yet decades worth of data reveal educator reports of maltreatment are the least likely to be screenedin and the least likely to be substantiated or confirmed. In other words, education personnel— whether motivated by genuine concern, which may nevertheless be informed by implicit biases towards low-income families and families of color; fear of liability; or the desire to access services they believe families cannot acquire elsewhere— overwhelm our child welfare system with unnecessary allegations of maltreatment. This reality has fundamentally transformed the relationship between families and schools. Carrying the heavy burden of mandated reporting laws, public schools disproportionately refer Black and low-income families to the family regulation system, abdicating schools’ opportunity to serve these same families in the communities in which they reside. Rather than serving as the great equalizer, public schools increasingly contribute to the carceral state’s regulation of families. This Article argues that schools must shift their role away from the reporting and surveillance of these families, and instead directly provide and arrange for services for families. This change begins with sharply limiting or repealing mandatory reporting obligations (permitting voluntary reports in severe cases)—but that is only the start. Schools are well-positioned to create new pathways to the supports and services from which most families reported to the family regulation system might actually benefit. Schools are already a primary source of food for impoverished children, and can help ensure low-income families access all the public benefits to which they are entitled. Schools can largely refer children and families to the same services that the family regulation system can—such as mental health services and substance abuse treatment—but without that system’s coercive authority and its associated problems. Where some services are tied to the family regulation system’s involvement, then law should permit schools to refer families directly. Schools know which families need legal services to defend their housing, access benefits, obtain orders of protection—or any of the myriad of other supports that poverty lawyers can provide. This shift would tie schools to the families and communities that they serve and benefit those families and communities far more than the surveillance and policing they experience under the current family regulation system.

How Racial Politics Led Directly to the Enactment of the Adoption and Safe Families Act of 1997

This Article is part of a celebration of the magnificent work of Dorothy Roberts who, more than any other scholar, has brilliantly demonstrated both the highly destructive qualities of the United States’ family regulation system and its relationship to the country’s legacy of slavery. The most vicious feature of the current family regulation system is the almost routine destruction of families resulting from an overly zealous enforcement of the Adoption and Safe Families Act of 1997, through which the federal government pays states to permanently banish parents from their children and legally sever the parent-child relationship when children have remained in foster care for fifteen months. This Article tells some of the racialized history that led to the enactment of the Adoption and Safe Families Act.

The Surveillance Tentacles of the Child Welfare System

The family regulation system identifies families through the use of widespread, cross-system surveillance for the purported purpose of keeping children safe. But the system does not surveil all families equally, leading to the disproportionate impact of family regulation on Black, Brown, and Native families, and fails to protect while causing more harm to children and communities of color. We examine how institutions and professionals that are meant to provide necessary services to the community—medical providers, social services agencies, the police, and schools—act as tentacles of surveillance, entrapping families in the family regulation system. We argue that engineering service and community providers as surveillance agents perpetuates inequality and leads to unnecessary family separation and trauma, and that genuine support for families can only thrive outside of the family regulation system and its surveillance tentacles.

Structural and mechanistic basis for protein glutamylation by the kinase fold

The kinase domain transfers phosphate from ATP to substrates. However, the Legionella effector SidJ adopts a kinase fold yet catalyzes calmodulin (CaM)-dependent glutamylation to inactivate the SidE ubiquitin ligases. The structural and mechanistic basis in which the kinase domain catalyzes protein glutamylation is unknown. Here we present cryo-EM reconstructions of SidJ:CaM:SidE reaction intermediate complexes. We show that the kinase-like active site of SidJ adenylates an active site Glu in SidE resulting in the formation of a stable reaction intermediate complex. An insertion in the catalytic loop of the kinase domain positions the donor Glu near the acyl-adenylate for peptide bond formation. Our structural analysis led us to discover that the SidJ paralog SdjA is a glutamylase that differentially regulates the SidE-ligases during Legionella infection. Our results uncover the structural and mechanistic basis in which the kinase fold catalyzes non-ribosomal amino acid ligations and reveal an unappreciated level of SidE-family regulation.

Family pedagogy and home education

The textbook reveals the normative and legislative acts of family regulation and the legal foundations of family education, describes the family at different stages of its formation, the history and traditions of the family in different faiths. The article presents the characteristics of family formation and marital relations. The basics of raising children in different types of families are formulated and methods of improving the pedagogical culture of parents are proposed. For students of secondary vocational education institutions. It can be useful for bachelors, undergraduates, postgraduates and students of advanced training courses and retraining of teaching staff.