The Molecular Mechanisms for Maintenance of Cancer Stem Cells in Chronic Myeloid Leukemia: A Dissertation

Chronic myeloid leukemia (CML) is a clonal hematopoietic stem cell disorder associated with the Philadelphia chromosome (Ph) that arises from a reciprocal translocation between chromosomes 9 and 22, thereby resulting in the formation of the chimeric BCR-ABL oncogene encoding a constitutively activated tyrosine kinase. BCR-ABL tyrosine kinase inhibitors (TKIs) induce a complete hematologic and cytogenetic response in the majority of chronic phrase CML patients. However, TKIs cannot efficiently eradicate leukemia stem cells (LSCs) because of the insensitivity of LSCs to TKIs. Therefore, developing new strategies to target LSCs is necessary and critical for curing CML, and success of this approach depends on further understanding the molecular mechanisms by which LSCs survive and are maintained. In Chapter I, I briefly introduce CML disease, BCR-ABL oncoprotein, and TKIs. I also describe the identification and features of LSCs. Several key pathways in LSCs including Wnt/ß-catenin, hedgehog, FoxO, Bcl6 and HIF1, are discussed. I also propose our strategy to identify unique molecular pathways that are important for LSCs but not their normal stem cell counterparts. In Chapter II, I describe our finding about the function of the positive regulator, HIF1α, in CML development and LSC survival. I show that loss of HIF1α impairs the maintenance of CML through impairing cell cycle progression and inducing apoptosis of LSCs, and I also report that p16Ink4a and p19Arf mediate the effect of HIF1α on LSCs, as knockdown of p16Ink4a and p19Arf rescues the defective colony-forming ability of HIF1α-/- LSCs. As detailed in Chapter III and IV, through comparing the global gene expression profiles of LSCs and HSCs, I find two novel regulators, Blk and Scd1, which act as tumor suppressors in CML development. In Chapter III, I show that Blk is markedly down-regulated by BCR-ABL in LSCs, and that c-Myc and Pax5 mediate this down-regulation. Deletion of Blk accelerates CML development; conversely, Blk overexpression significantly delays the development of CML and impairs the function of LSCs. I also demonstrate that p27, as a downstream effector, is involved in the function of Blk in LSCs. Blk also functions as a tumor suppressor in human CML stem cells, and inhibits the colony-forming ability of human CML cells. In Chapter IV, I investigate the function of another negative regulator, Scd1, in CML LSCs, and find that expression of Scd1 is down-regulated in mouse LSCs and human CML cells. We report that Scd1 acts as a tumor suppressor in CML, as loss of Scd1 causes acceleration of CML development and overexpression of Scd1 delays CML development. Using a colony-forming assay, I demonstrate that Scd1 impairs the maintenance of LSCs due to the change of expression of Pten, p53 and Bcl2. Importantly, I find that both Blk and Scd1 do not affect normal hematopoietic stem cells (HSCs) or hematopoiesis. Taken together, our findings demonstrate that HIF1α is required for the maintenance of CML LSCs, and conversely that Blk and Scd1 suppress the function of LSCs, suggesting that combining TKI treatment with specific targeting of LSCs will be necessary for curing CML

Exploiting Unique Biological Features of Leukemia Stem Cells for Therapeutic Benefit

Cancer stem cells play a critical role in disease initiation and insensitivity to chemotherapy in numerous hematologic malignancies and some solid tumors, and these stem cells need to be eradicated to achieve a cure. Key to successful targeting of cancer stem cells is to identify and functionally test critical target genes and to fully understand their associated molecular network in these stem cells. Human chronic myeloid leukemia (CML) is well accepted as one of the typical types of hematopoietic malignancies that are derived from leukemia stem cells (LSCs), serving as an excellent model disease for understanding the biology of LSCs and developing effective, selective, and curative strategies through targeting LSCs. Here, we discuss LSCs in CML with a focus on identification of unique biological features of these stem cells to emphasize the feasibility and significance of specific targeting of LSCs while sparing normal stem cell counterparts in leukemia therapy

Solubility behavior and thermodynamic modeling of inosine (form β) in four cosolvency systems at T = 278.15 to 323.15 K

N,N-Dimethylformamide (DMF) as the main solvent with strong dissolving power was blended with four secondary solvents (ethanol, n-propanol, isopropanol, and propylene glycol) with relatively weak dissolving power to form many new solvents. The dispersion index (inosine (form β) mole fraction) in five organic solvents such as DMF and in the newly formed solvent was also obtained one by one with the static method commonly used in solid–liquid equilibrium. The temperature environment includes the high temperature set at 323.15 K, the low temperature set at 278.15 K, and an interval between each temperature of 5 K. The pressure environment was the atmospheric pressure in the natural state, and the usual value was 101.0 kPa. In a mixed system, temperature was a non-negligible influencing factor from beginning to end, and its increase often led the solute to the trend of high solubility. In addition, the proportion of the main solvent also dominated the solubilization trend of the inosine (form β); the larger the proportion of DMF, the easier the dissolution process. When both of the above factors were fixed at a certain point, the dispersing ability of the dispersing liquid composed of DMF and ethanol was undoubtedly the first. Three models (Jouyban–Acree model, van’t Hoff–Jouyban–Acree model, and modified Apelblat–Jouyban–Acree model) were used to correlate the solubility data. The largest relative average deviation and root-mean-square deviation values were 4.66 × 10–2 and 7.27 × 10–4, respectively. The dispersion data of inosine (form β) obtained through this experimental process and related thermodynamic parameters obtained through thermodynamic calculations have important application significance for its industrial production and further purification

An Efficient Multi-solution Solver for the Inverse Kinematics of 3-Section Constant-Curvature Robots

Piecewise constant curvature is a popular kinematics framework for continuum robots. Computing the model parameters from the desired end pose, known as the inverse kinematics problem, is fundamental in manipulation, tracking and planning tasks. In this paper, we propose an efficient multi-solution solver to address the inverse kinematics problem of 3-section constant-curvature robots by bridging both the theoretical reduction and numerical correction. We derive analytical conditions to simplify the original problem into a one-dimensional problem. Further, the equivalence of the two problems is formalised. In addition, we introduce an approximation with bounded error so that the one dimension becomes traversable while the remaining parameters analytically solvable. With the theoretical results, the global search and numerical correction are employed to implement the solver. The experiments validate the better efficiency and higher success rate of our solver than the numerical methods when one solution is required, and demonstrate the ability of obtaining multiple solutions with optimal path planning in a space with obstacles.Comment: Robotics: Science and Systems 202

Experimental determination and thermodynamic correlation of 7-amino-4-methylcoumarin solubility in various cosolvency mixtures at (278.15–323.15) K

Four solvents (ethanol, isopropanol, ethylene glycol (EG), and N,N-dimethylformamide (DMF)) that can be mixed with water in any ratio were selected to determine the dissolution performance of 7-amino-4-methylcoumarin by the classical shake-flask method. The measured temperature range began at 278.15 K and ended at 303.15 K, and the pressure environment was controlled at standard atmospheric pressure (101.1 kPa). Results reveal that with the addition of organic solvents, the solubilization effect of 7-amino-4-methylcoumarin was very significant and the larger the amount of addition, the more obvious was the effect of solubilization. Not only that, the temperature change had a non-negligible effect on the dispersion of 7-amino-4-methylcoumarin; the temperature increased monotonically, and the better was the dissolution. When the external conditions were kept constant, the addition of DMF made the solubilization effect of 7-amino-4-methylcoumarin most obvious among all organic solvents used. This study involved three models, including the Jouyban–Acree model and its two variants (van’t Hoff–Jouyban–Acree model and Apelblat–Jouyban–Acree model), which were used to correlate the solubility data of 7-amino-4-methylcoumarin in aqueous cosolvent mixtures. The relative average deviation (RAD) and root-mean-square deviation (RMSD) reaches to 3.47 × 10–2 and 1.79 × 10–3 rooting in the van’t Hoff–Jouyban–Acree model. The relevant parameters obtained through model calculation and experimental means are essential for product synthesis, separation, and purification processes

Prompt Tuning with Soft Context Sharing for Vision-Language Models

Vision-language models have recently shown great potential on many computer vision tasks. Meanwhile, prior work demonstrates prompt tuning designed for vision-language models could acquire superior performance on few-shot image recognition compared to linear probe, a strong baseline. In real-world applications, many few-shot tasks are correlated, particularly in a specialized area. However, such information is ignored by previous work. Inspired by the fact that modeling task relationships by multi-task learning can usually boost performance, we propose a novel method SoftCPT (Soft Context Sharing for Prompt Tuning) to fine-tune pre-trained vision-language models on multiple target few-shot tasks, simultaneously. Specifically, we design a task-shared meta network to generate prompt vector for each task using pre-defined task name together with a learnable meta prompt as input. As such, the prompt vectors of all tasks will be shared in a soft manner. The parameters of this shared meta network as well as the meta prompt vector are tuned on the joint training set of all target tasks. Extensive experiments on three multi-task few-shot datasets show that SoftCPT outperforms the representative single-task prompt tuning method CoOp [78] by a large margin, implying the effectiveness of multi-task learning in vision-language prompt tuning. The source code and data will be made publicly available

GABP transcription factor is required for development of chronic myelogenous leukemia via its control of PRKD2

Hematopoietic stem cells (HSCs) are the source of all blood lineages, and HSCs must balance quiescence, self-renewal, and differentiation to meet lifelong needs for blood cell development. Transformation of HSCs by the breakpoint cluster region-ABL tyrosine kinase (BCR-ABL) oncogene causes chronic myelogenous leukemia (CML). The E-twenty six (ets) transcription factor GA binding protein (GABP) is a tetrameric transcription factor complex that contains GABPalpha and GABPbeta proteins. Deletion in bone marrow of Gabpa, the gene that encodes the DNA-binding component, caused cell cycle arrest in HSCs and profound loss of hematopoietic progenitor cells. Loss of Gabpalpha prevented development of CML, although mice continued to generate BCR-ABL-expressing Gabpalpha-null cells for months that were serially transplantable and contributed to all lineages in secondary recipients. A bioinformatic screen identified the serine-threonine kinase protein kinase D2 (PRKD2) as a potential effector of GABP in HSCs. Prkd2 expression was markedly reduced in Gabpalpha-null HSCs and progenitor cells. Reduced expression of PRKD2 or pharmacologic inhibition decreased cell cycling, and PRKD2 rescued growth of Gabpalpha-null BCR-ABL-expressing cells. Thus, GABP is required for HSC cell cycle entry and CML development through its control of PRKD2. This offers a potential therapeutic target in leukemia

The Blk pathway functions as a tumor suppressor in chronic myeloid leukemia stem cells

A therapeutic strategy for treating cancer is to target and eradicate cancer stem cells (CSCs) without harming their normal stem cell counterparts. The success of this approach relies on identification of molecular pathways that selectively regulate CSC function. Using BCR-ABL-induced chronic myeloid leukemia (CML) as a disease model for CSCs, we show that BCR-ABL down-regulates the B lymphoid kinase (Blk) gene through c-Myc in leukemia stem cells (LSCs) in CML mice and that Blk functions as a tumor suppressor in LSCs but does not affect normal hematopoietic stem cells (HSCs) or hematopoiesis. Blk suppresses LSC function through a pathway involving an upstream regulator, Pax5, and a downstream effector, p27. Inhibition of this Blk pathway accelerates CML development, whereas increased activity of the Blk pathway delays CML development. Blk also suppresses human CML stem cells. Our results demonstrate the feasibility of selectively targeting LSCs, an approach that should be applicable to other cancers

LSK Derived LSK– Cells Have a High Apoptotic Rate Related to Survival Regulation of Hematopoietic and Leukemic Stem Cells

A balanced pool of hematopoietic stem cells (HSCs) in bone marrow is tightly regulated, and this regulation is disturbed in hematopoietic malignancies such as chronic myeloid leukemia (CML). The underlying mechanisms are largely unknown. Here we show that the Lin−Sca-1+c-Kit- (LSK−) cell population derived from HSC-containing Lin−Sca-1+c-Kit+ (LSK) cells has significantly higher numbers of apoptotic cells. Depletion of LSK cells by radiation or the cytotoxic chemical 5-fluorouracil results in an expansion of the LSK− population. In contrast, the LSK− population is reduced in CML mice, and depletion of leukemia stem cells (LSCs; BCR-ABL-expressing HSCs) by deleting Alox5 or by inhibiting heat shock protein 90 causes an increase in this LSK− population. The transition of LSK to LSK− cells is controlled by the Icsbp gene and its downstream gene Lyn, and regulation of this cellular transition is critical for the survival of normal LSK cells and LSCs. These results indicate a potential function of the LSK− cells in the regulation of LSK cells and LSCs