**4. Conclusions**

**Author Contributions:** Conceptualization, C.S. and G.F.; validation, C.S., G.F. and X.M.; formal analysis, C.S. and G.F.; and writing—original draft preparation, G.F.; writing—review and editing, C.S.; funding acquisition, D.M. All authors have read and agreed to the published version of the manuscript.

**Funding:** The authors would like to acknowledge the financial assistance provided by Project of State Key Laboratory of High Performance Complex Manufacturing, Central South University (No.ZZYJKT2019-09), and National Basic Research Program of China (No.2014CB046702).

**Conflicts of Interest:** The authors declare no conflict of interest.


**5. Conclusions**

272 Advances in Hematopoietic Stem Cell Research

acute leukemia in adults, affecting roughly three out of 100.000 people. AML patients are predominantly elderly, with a median age at diagnosis of 67 (National Cancer Institute

In AML, there is wide patient-to-patient heterogeneity in the appearance of the leukemic blasts. Conventionally, AML is classified into seven French–American–British (FAB)

The discovery of leukemia-initiating cells in acute myeloid leukemias (AMLs) started with the discovery that the large majority of AML blasts do not proliferate and only a small

A common feature to all AML cases is the arrested aberrant differentiation leading to an accumulation of more than 20% blast cells in the bone marrow (Gilliland & Tallman, 2002). More than 80% of myeloid leukemias are associated with at least one chromosomal rearrangement (Pandolfi, 2001), and over 100 different chromosomal translocations have been cloned (Gilliland & Tallman, 2002). Frequently, these translocations involve genes encoding transcription factors that have been shown to play an important role in hematopoietic lineage development. Thus, alteration of the transcriptional machinery appears to be a common

Clinical investigation and experimental animal models suggest that at least two genetic alterations are required for the clinical manifestation of acute leukemia. According to the model proposed by Gilliland & Tallman (2002), cooperation between class I activating mutations and class II mutations that induce termination of differentiation give rise to AML. The class I mutations, such as mutations in the receptor tyrosine kinase genes FLT3 and KIT, RAS family members, and loss of function of neurofibromin 1, confer proliferative and/or survival advantage to hematopoietic progenitors, typically as a consequence of aberrant activation of signal transduction pathways. The class II mutations lead to a halt in differentiation via

While the LSC appears to share many of the cell surface markers previously identified for HSC such as CD34, CD38, HLA-DR, and CD71, there have been several groups who have reported surface markers that are differentially expressed in the two populations. CD90 or Thy-1 is one marker that has been described to be potentially specific of the LSC compartment. Thy-1 is downregulated in normal hematopoiesis as the most primitive stem cells progress toward the progenitor stage. This finding of the lack of expression on LSC might suggest that the primitive stem cell does not contribute to the primary pathological event, or that Thy-1 expression is downregulated as a result of the leukemogenic events

The interaction between CXCL12 (stromal cell–derived factor-1 alpha) and its receptor CXCR4 on leukemic progenitor cells contributes to their homing to the bone marrow microenvironment. CXCR4 levels are significantly elevated in leukemic cells from patients with AML and CXCR4 expression is associated with poor outcome (Konopleva & Jordan, 2011). Constitutive activation of the nuclear factor kappa B (NF-kB) pathway in primary human AML stem cells provided evidence that NF-kB plays a significant role in the overall survival of LSCs as well as AML cell types in general. This pathway is strongly implicated

as a central target in developing LSC-specific therapies (Konopleva & Jordan, 2011).

subtypes corresponding to the maturation stage of the leukemia (Warner et al., 2004).

mechanism leading to arrested differentiation (Pandolfi, 2001; Tenen, 2003).

interference with transcription factors or co-activators (Frankfurt et al., 2009).

minority is capable of forming new colonies (Testa, 2011).

1975–2007).

(Hope et al., 2003).

Adult hematopoietic stem cells are undifferentiated cells capable of self-renewal and differentiation potential in several cell types that comprise the hematopoietic tissue. These cells have been used in bone marrow transplantation for treatment of hematological malignancies as well as non-hematological diseases. The HSC is the main component in the process of hematopoiesis which, together with the cells that make up the bone marrow stromal environment and other intrinsic and extrinsic factors, orchestrates the entire production of progenitors and terminally differentiated blood cells.

However, when this process of cell production is unbalanced, leading to an exacerbated and uncontrolled proliferation of blood progenitor cells, leukemia may develop.The ultimate challenge in coming years will be to understand the stem cell 'programme', particularly the control of self-renewal, in an attempt to develop novel, stem cell-directed therapies. An improved understanding of clonal evolution will be critical if we are to ensure that cancers are not able to evolve mechanisms to evade the new directed therapies. However, reducing the risk of relapse and minimizing long-term side effects should always remain the ultimate goal of understanding the CSCs.

With little doubt, the leukemia stem cell model has had the greatest clinical impact on our understanding and treatment of Philadelphia chromosome positive leukemias. Effective

Hematopoietic Stem Cell in Acute Myeloid Leukemia Development 275

Gordon, M. *Stem cells and haemopoiesis*. In: Hoffbrand, V., Catovsky, D., Tuddenham, E.G., 5th

Guzman ML, Neering SJ, Upchurch D, Grimes B, Howard DS, Rizzieri DA, Luger SM,

Hanahan, D. & Weinberg, R.A. (2000). The hallmarks of cancer*. Cell* Vol.100, No. 1, (January

Hope, K. J., Jin, L., Dick, JE. (2003). Human Acute Myeloid Leukemia Stem Cells*. Archives of Medical Research* Vol.34, No. 6, (November-December 2003), pp. 507–514. Hosen, N., Park, C.Y., Tatsumi, N., Oji, Y., Sugiyama, H., Gramatzki, M., Krensky, A.M.,

Huls, M., Russel, F.G., Masereeuw, R. (2009). The role of ATP binding cassette transporters

Jing, D., Fonseca, A.V., Alakel, N., Fierro, F.A., Muller, K., Bornhauser, M., Ehninger, G.,

Kohler, B.A., Ward, E., McCarthy, B.J., Schymura, M.J., Ries, L.A., Eheman, C., Jemal, A.,

Konopleva, M.Y. & Jordan, CT*.* (2011). Leukemia Stem Cells and Microenvironment. *Biology*

Lane, S.W., Wang, Y.J., Lo Celso, C., Ragu, C., Bullinger, L., Sykes, S.M., Ferraro, F.,

Lapidot, T, Sirard, C., Vormoor, J., Murdoch, B., Hoang, T., Caceres-Cortes, J., Minden, M.,

Marques, D.S., Sandrini, J.Z., Boyle, R.T., Marins, L.F., Trindade, G.S. (2010). Relationships

Pandolfi, P.P. (2001). In vivo analysis of the molecular genetics of acute promyelocytic leukemia. *Oncogene* Vol.20, No. 40, (September 2001), pp. 5726-5735. Pollyea, D.A., Kohrt, H.E., Medeiros, B.C. (2011). Acute myeloid leukaemia in the elderly: a review. *British Journal of Haematology* Vol.152, No. 5, (March 2011), pp. 524-542.

*and Therapeutic Targeting* Vol.29, No. 5, (May 2011), pp. 591-599.

leukemia progression. *Blood* (July 2011), *in press.*

(February 1994), pp. 645–648.

acute myeloid leukemia. pp. 1-12. New York.

26, (June 2007), pp. 11008-11013.

*Therapy* Vol.328, No. 1, (January 2009), pp. 3-9.

*Haematologica* Vol.95, No. 4, (April 2010), pp. 542550.

2000), 57-70.

37-46.

762.

acute myelogenous leukemia cells. *Blood*: 2301–2307.

ed. Blackwell Publishing, (2005): Differential niche and Wnt requirements during

Jordan CT (2001). Nuclear factor-KB is constitutively activated in primitive human

Weissman, I.L. (2007). CD96 is a leukemic stem cell-specific marker in human acute myeloid leukemia. *Proceedings of National Academy of Science of U S A* Vol.104, No.

in tissue defense and organ regeneration. *Journal of Pharmacogycal Experimental*

Corbeil, D., Ordemann, R. (2010). Hematopoietic stem cells in co-culture with mesenchymal stromal cells-modeling the niche compartments in vitro.

Anderson, R.N., Ajani, U.A., Edwards, B.K. (2011). Annual report to the nation on the status of cancer, 1975-2007, featuring tumors of the brain and other nervous system*. Journal of National Cancer Institute* Vol.103, No. 9, (May 2011), pp. 714-736. Kondo, M. (2010). Lymphoid and myeloid lineage commitment in multipotent

hematopoietic progenitors. *Immunology Reviews* Vol. 238, No. 1, (January 2010), pp.

Shterental, S., Lin, C.P., Gilliland, D.G., Scadden, D.T., Armstrong, S.A., Williams, D.A. (2011). Differential niche and Wnt requirements during acute myeloid

Paterson, B., Caliguri, M.A., Dick, J.E. (1994). A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. *Nature* Vol.367, No. 6464,

between multidrug resistance (MDR) and stem cell markers in human chronic myeloid leukemia cell lines*. Leukemia Research* Vol.34, No. 6, (June 2010), pp. 757–

targeted agents and the ability to follow the impact of therapy on critical rare subpopulations of the malignant clone has greatly advanced our understanding of chronic myeloid leukemia.

For AML, the clinical impact of the leukemic stem cell model is less clear. The ability to isolate and characterize rare LSC populations has had a significant importance on our understanding of the biology of AML. In the past decade, we have gained considerable insight into the properties that distinguish leukemic stem cells from their normal counterparts and some of the rules that govern the leukemic hierarchy.

Despite the wide variance in techniques and to some degree expression profiles, common signaling pathways have been shown to play a role not only in AML stem cells, but also in cancer stem cells in general. These include BMI1, Wnt, Sonic Hedgehog, Notch, and NF-kB. These pathways are being evaluated for their role in LSC biology and agents targeting these pathways are making their way through the pre-clinical focus.

### **6. References**

targeted agents and the ability to follow the impact of therapy on critical rare subpopulations of the malignant clone has greatly advanced our understanding of chronic

For AML, the clinical impact of the leukemic stem cell model is less clear. The ability to isolate and characterize rare LSC populations has had a significant importance on our understanding of the biology of AML. In the past decade, we have gained considerable insight into the properties that distinguish leukemic stem cells from their normal

Despite the wide variance in techniques and to some degree expression profiles, common signaling pathways have been shown to play a role not only in AML stem cells, but also in cancer stem cells in general. These include BMI1, Wnt, Sonic Hedgehog, Notch, and NF-kB. These pathways are being evaluated for their role in LSC biology and agents targeting these

Becker, M. W. & Jordan, C. T. (2011). Leukemia stem cells in 2010: Current understanding and future directions. *Blood Reviews* Vol.25, No. 2, (March 2011), pp. 75–81. Blair, A.;Hogge, D.E. & Sutherland, H.J. (1998). Most acute myeloid leukemia progenitor

Bomken, S., Fiser, K., Heidenreich, O. (2010). Vormoor, J. Understanding the cancer stem cell. *British Journal of Cancer* Vol.103, No. 4, (August 2010), pp. 439 – 445. Broner, F. & Carson, M C. *Topics in bone biology*. Springer. 2009; 4: pp. 2-4. New York, USA. Buting, K. D. (2002). ABC transporters as phenotypic markers and functional regulators of

Cao, R., Tsukada, Y., Zhang, Y. (2005). Role of Bmi-1 and Ring1A in H2A ubiquitylation and Hox gene silencing. *Molecular Cell* Vol.22, No. 6, (December 2005),pp. 845-854. Chen, Y.C., Hsu, H.S., Chen, Y.W., Tsai, T.H., How, C.K., Wang, C.Y.,

Dick, J.E. (2003). Stem cells: Self-renewal writ in blood. *Nature* Vol.423, No. 6937, (May 2003),

Du, W., Adam, Z., Rani, R., Zhang, X., Pang, Q. (2008). Oxidative stress in Fanconi anemia

Frankfurt, O., Licht, J.D., Tallman, M.S. (2007). Molecular characterization of acute myeloid

Gilliland, D.G.& Tallman, M.S. (2002). Focus on acute leukemias. *Cancer Cell.* Vol.1, No. 5,

Hung,S.C.,Chang,Y.L.,Tsai,M.L.,Lee,Y.Y.,Ku,H.H.,Chiou,S.H.(2008). Oct-4 expression maintained cancer stem-like properties in lung cancer-derived CD-133

hematopoiesis and disease progression. *Antioxidants & Redox Signaling* Vol.10, No.

leukemia and its impact on treatment. *Current Opinion in Oncology* Vol.19, No. 6,

stem cells. *Stem Cells* Vol.20, No. 12, (December 2002), pp. 11-20.

positive cells. *PLoS ONE* Vol.3, No. 7, (July 2008), pp. 2637.

cells with long-term proliferative ability in vitro and in vivo have the phenotype CD34þ/CD71-/HLA-DR-. *Blood* Vol.92, No. 11, (December 1998), pp. 4325–4335. Bojanić, I. & Golubić Cepulić, B. (2006).Umbilical cord blood as a source of stem cells. *Acta*

counterparts and some of the rules that govern the leukemic hierarchy.

*Medica Croatica* Vol.60, No. 3, (June 2006),pp. 215-225.

pathways are making their way through the pre-clinical focus.

myeloid leukemia.

**6. References**

pp. 231–233.

11 (November 2008), pp. 1909-1921.

(November 2007), pp. 635–649.

(June 2002), pp. 417-420.

**13**

*Mexico*

**From HSC to B-Lymphoid Cells in Normal and**

**Malignant Hematopoiesis**

Rosana Pelayo1, Elisa Dorantes-Acosta1,2,

 *Mexican Institute for Social Security, Mexico City*

*Social Security, Mexico City*

Eduardo Vadillo1 and Ezequiel Fuentes-Pananá3

*1Oncology Research Unit, Oncology Hospital, Mexican Institute for*

*2Leukemia Clinic, Mexican Children's Hospital 'Federico Gómez', Mexico City 3Research Unit on Parasitic and Infectious Diseases, Pediatric Hospital,*

Development of B-lymphoid cells is a highly ordered multi-step process that, in adult mammals, starts in bone marrow in a pool of self-renewing multipotential hematopoietic stem cells, which gradually commit to the lymphoid lineage and advance through high regulated differentiation pathways until formation of mature functional cells. Over the last few years, exceptional advances have been recorded in identifying primitive progenitors that lay the foundations of the lymphoid program while losing myeloid potential, along with patterns of transcriptional activity controlling lineage fate decisions and environmental cues that influence the differentiation pathway during normal hematopoiesis. Multicolor flow cytometry, controlled cell cultures, genetic marking systems, microarray technologies and xenotransplantation approaches are being extensively used to address fundamental questions on this regard. Of special interest is the stem cell research with relevance to hierarchy and early events in malignant lymphopoiesis, and to new insights into perspectives that may allow progress in means to protect and sustain the immune system during chemotherapy, inflammation, infection, and following hematopoietic transplantation. In this book chapter, we focus on the hierarchical structure of the early lymphoid system, the current knowledge about intrinsic and microenvironmental factors regulating the differentiation of lymphoid progenitors, and the emerging research to understand malignant lymphoid

Mature blood cells are constantly replaced from a unique cell population of hematopoietic stem cells (HSC) residing in specialized niches within the bone marrow (BM), where the hematopoietic system is organized as a hierarchy of cell types that gradually lose multiple alternate potentials while commit to lineage fates and gain specialized functions (Baba et

**1. Introduction**

development.

**2. The early steps in the lymphoid development**


**4.2 Container sewage wastewater treatment plant (CSWTP)—leachate treatment plant in reverse osmosis technology**

Container sewage wastewater treatment plant (CSWTP) with a capacity of 30 m3 /d includes system called a single-stage membrane process ensuring obtaining the leachate parameters enabling them to be safely transported to the municipal sewage treatment plant. The treatment plant operates on the basis of the reverse osmosis process (e.g., **Figure 6**), the essence of which consists in passing the leachate from the storage site through a semipermeable membrane under the influence of the pressure set on the inlet side of the effluent.

Membrane separation is a purely physical separation; separated components do not undergo any chemical or biological transformation. The applied solution is a pilot solution for cooperation with the existing pretreatment plant in order to increase the cleaning effects. The effect of the treatment plant is to obtain purified leachate (permeate) and leachate residue (concentrate).

The sewage treatment plant works in a continuous system and cleanses the effluents from the ZRO2 reservoir, i.e., from quarters 9 to 12. The treatment plant will be controlled by means of a computer program and will work in an automatic system. The computer will monitor, through systematic conductivity measurement, the quality of treated leachate discharged into the environment. If the conductivity

**Figure 6.**  *The treatment plant based on the reverse osmosis process (source: [18]).*

rises above the programmed value, the installation will automatically stop, and a cleaning program for filters or modules will start.

Sewage plant is located in a paved square for turning vehicles. Container of treatment plant with dimensions of 12.2 × 2.5 m is set on a separate foundation.

The container is made of steel construction with a layer casing made of trapezoid sheet metal from the outside and a polypropylene plate from the inside of the container. The addition between the layers is a mineral wool insulation layer. Tight floor is made of chemically resistant material. The container has mechanical ventilation.