Increased heterogeneity in expression of genes associated with cancer progression and drug resistance.

Fluctuations in the number of regulatory molecules and differences in timings of molecular events can generate variation in gene expression among genetically identical cells in the same environmental condition. This variation, termed as expression noise, can create differences in metabolic state and cellular functions, leading to phenotypic heterogeneity. Expression noise and phenotypic heterogeneity have been recognized as important contributors to intra-tumor heterogeneity, and have been associated with cancer growth, progression, and therapy resistance. However, how expression noise changes with cancer progression in actual cancer patients has remained poorly explored. Such an analysis, through identification of genes with increasing expression noise, can provide valuable insights into generation of intra-tumor heterogeneity, and could have important implications for understanding immune-suppression, drug tolerance and therapy resistance. In this work, we performed a genome-wide identification of changes in gene expression noise with cancer progression using single-cell RNA-seq data of lung adenocarcinoma patients at different stages of cancer. We identified 37 genes in epithelial cells that showed an increasing noise trend with cancer progression, many of which were also associated with cancer growth, EMT and therapy resistance. We found that expression of several of these genes was positively associated with expression of mitochondrial genes, suggesting an important role of mitochondria in generation of heterogeneity. In addition, we uncovered substantial differences in sample-specific noise profiles which could have implications for personalized prognosis and treatment.

KDM6 demethylases integrate DNA repair gene regulation and loss of KDM6A sensitizes human acute myeloid leukemia to PARP and BCL2 inhibition.

Acute myeloid leukemia (AML) is a heterogeneous, aggressive malignancy with dismal prognosis and with limited availability of targeted therapies. Epigenetic deregulation contributes to AML pathogenesis. KDM6 proteins are histone-3-lysine-27-demethylases that play context-dependent roles in AML. We inform that KDM6-demethylase function critically regulates DNA-damage-repair-(DDR) gene expression in AML. Mechanistically, KDM6 expression is regulated by genotoxic stress, with deficiency of KDM6A-(UTX) and KDM6B-(JMJD3) impairing DDR transcriptional activation and compromising repair potential. Acquired KDM6A loss-of-function mutations are implicated in chemoresistance, although a significant percentage of relapsed-AML has upregulated KDM6A. Olaparib treatment reduced engraftment of KDM6A-mutant-AML-patient-derived xenografts, highlighting synthetic lethality using Poly-(ADP-ribose)-polymerase-(PARP)-inhibition. Crucially, a higher KDM6A expression is correlated with venetoclax tolerance. Loss of KDM6A increased mitochondrial activity, BCL2 expression, and sensitized AML cells to venetoclax. Additionally, BCL2A1 associates with venetoclax resistance, and KDM6A loss was accompanied with a downregulated BCL2A1. Corroborating these results, dual targeting of PARP and BCL2 was superior to PARP or BCL2 inhibitor monotherapy in inducing AML apoptosis, and primary AML cells carrying KDM6A-domain mutations were even more sensitive to the combination. Together, our study illustrates a mechanistic rationale in support of a novel combination therapy for AML based on subtype-heterogeneity, and establishes KDM6A as a molecular regulator for determining therapeutic efficacy.

Self-stresses control stiffness and stability in overconstrained disordered networks.

We investigate the interplay between prestress and mechanical properties in random elastic networks. To do this in a controlled fashion, we introduce an algorithm for creating random free-standing frames that support exactly one state of self-stress. By multiplying all the bond tensions in this state of self-stress by the same number-which with the appropriate normalization corresponds to the physical prestress inside the frame-we systematically evaluate the linear mechanical response of the frame as a function of prestress. After proving that the mechanical moduli of affinely deforming frames are rigorously independent of prestress, we turn to nonaffinely deforming frames. In such frames, prestress has a profound effect on linear response: not only can it change the values of the linear modulus-an effect we demonstrate to be related to a suppressive effect of prestress on nonaffinity-but prestresses also generically trigger a bistable mechanical response. Thus, prestress can be leveraged to both augment the mechanical response of network architectures on the fly, and to actuate finite deformations. These control modalities may be of use in the design of both novel responsive materials and soft actuators.

Geometry and the onset of rigidity in a disordered network.

Disordered spring networks that are undercoordinated may abruptly rigidify when sufficient strain is applied. Since the deformation in response to applied strain does not change the generic quantifiers of network architecture, the number of nodes and the number of bonds between them, this rigidity transition must have a geometric origin. Naive, degree-of-freedom-based mechanical analyses such as the Maxwell-Calladine count or the pebble game algorithm overlook such geometric rigidity transitions and offer no means of predicting or characterizing them. We apply tools that were developed for the topological analysis of zero modes and states of self-stress on regular lattices to two-dimensional random spring networks and demonstrate that the onset of rigidity, at a finite simple shear strain γ^{★}, coincides with the appearance of a single state of self-stress, accompanied by a single floppy mode. The process conserves the topologically invariant difference between the number of zero modes and the number of states of self-stress but imparts a finite shear modulus to the spring network. Beyond the critical shear, the network acquires a highly anisotropic elastic modulus, resisting further deformation most strongly in the direction of the rigidifying shear. We confirm previously reported critical scaling of the corresponding differential shear modulus. In the subcritical regime, a singular value decomposition of the network's compatibility matrix foreshadows the onset of rigidity by way of a continuously vanishing singular value corresponding to the nascent state of self-stress.